TY  - JOUR
AU  - Levent, Gizem
AU  - Božić, Aleksandar
AU  - Petrujkić, Branko
AU  - Callaway, Todd R.
AU  - Poole, Toni L.
AU  - Crippen, Tawni L.
AU  - Harvey, Roger B.
AU  - Ochoa-García, Pedro
AU  - Corral-Luna, Agustin
AU  - Yeater, Kathleen M.
AU  - Anderson, Robin C.
PY  - 2024
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/3703
AB  - Livestock producers need new technologies to maintain the optimal health and wellbeing of their animals while minimizing the risks of propagating and disseminating pathogenic and
antimicrobial-resistant bacteria to humans or other animals. Where possible, these interventions
should contribute to the efficiency and profitability of animal production to avoid passing costs on
to consumers. In this study, we examined the potential of nitroethane, 3-nitro-1-propionate, ethyl
nitroacetate, taurine and L-cysteinesulfinic acid to modulate rumen methane production, a digestive
inefficiency that results in the loss of up to 12% of the host’s dietary energy intake and a major
contributor of methane as a greenhouse gas to the atmosphere. The potential for these compounds
to inhibit the foodborne pathogens, Escherichia coli O157:H7 and Salmonella Typhimurium DT104,
was also tested. The results from the present study revealed that anaerobically grown O157:H7 and
DT104 treated with the methanogenic inhibitor, ethyl nitroacetate, at concentrations of 3 and 9 mM
had decreased (p < 0.05) mean specific growth rates of O157:H7 (by 22 to 36%) and of DT104 (by 16 to
26%) when compared to controls (0.823 and 0.886 h−1
, respectively). The growth rates of O157:H7 and
DT104 were decreased (p < 0.05) from controls by 31 to 73% and by 41 to 78% by α-lipoic acid, which
we also found to inhibit in vitro rumen methanogenesis up to 66% (p < 0.05). Ethyl nitroacetate was
mainly bacteriostatic, whereas 9 mM α-lipoic acid decreased (p < 0.05) maximal optical densities
(measured at 600 nm) of O157:H7 and DT104 by 25 and 42% compared to controls (0.448 and 0.451,
respectively). In the present study, the other oxidized nitro and organosulfur compounds were neither
antimicrobial nor anti-methanogenic.
PB  - MDPI
T2  - Microorganisms
T1  - Assessment of potential anti-methanogenic and antimicrobial activity of ethyl nitroacetate, α-lipoic acid, taurine and L-cysteinesulfinic acid In vitro
VL  - 12
IS  - 1
SP  - 34
DO  - 10.3390/microorganisms12010034
ER  - 

@article{
author = "Levent, Gizem and Božić, Aleksandar and Petrujkić, Branko and Callaway, Todd R. and Poole, Toni L. and Crippen, Tawni L. and Harvey, Roger B. and Ochoa-García, Pedro and Corral-Luna, Agustin and Yeater, Kathleen M. and Anderson, Robin C.",
year = "2024",
abstract = "Livestock producers need new technologies to maintain the optimal health and wellbeing of their animals while minimizing the risks of propagating and disseminating pathogenic and
antimicrobial-resistant bacteria to humans or other animals. Where possible, these interventions
should contribute to the efficiency and profitability of animal production to avoid passing costs on
to consumers. In this study, we examined the potential of nitroethane, 3-nitro-1-propionate, ethyl
nitroacetate, taurine and L-cysteinesulfinic acid to modulate rumen methane production, a digestive
inefficiency that results in the loss of up to 12% of the host’s dietary energy intake and a major
contributor of methane as a greenhouse gas to the atmosphere. The potential for these compounds
to inhibit the foodborne pathogens, Escherichia coli O157:H7 and Salmonella Typhimurium DT104,
was also tested. The results from the present study revealed that anaerobically grown O157:H7 and
DT104 treated with the methanogenic inhibitor, ethyl nitroacetate, at concentrations of 3 and 9 mM
had decreased (p < 0.05) mean specific growth rates of O157:H7 (by 22 to 36%) and of DT104 (by 16 to
26%) when compared to controls (0.823 and 0.886 h−1
, respectively). The growth rates of O157:H7 and
DT104 were decreased (p < 0.05) from controls by 31 to 73% and by 41 to 78% by α-lipoic acid, which
we also found to inhibit in vitro rumen methanogenesis up to 66% (p < 0.05). Ethyl nitroacetate was
mainly bacteriostatic, whereas 9 mM α-lipoic acid decreased (p < 0.05) maximal optical densities
(measured at 600 nm) of O157:H7 and DT104 by 25 and 42% compared to controls (0.448 and 0.451,
respectively). In the present study, the other oxidized nitro and organosulfur compounds were neither
antimicrobial nor anti-methanogenic.",
publisher = "MDPI",
journal = "Microorganisms",
title = "Assessment of potential anti-methanogenic and antimicrobial activity of ethyl nitroacetate, α-lipoic acid, taurine and L-cysteinesulfinic acid In vitro",
volume = "12",
number = "1",
pages = "34",
doi = "10.3390/microorganisms12010034"
}

Levent, G., Božić, A., Petrujkić, B., Callaway, T. R., Poole, T. L., Crippen, T. L., Harvey, R. B., Ochoa-García, P., Corral-Luna, A., Yeater, K. M.,& Anderson, R. C.. (2024). Assessment of potential anti-methanogenic and antimicrobial activity of ethyl nitroacetate, α-lipoic acid, taurine and L-cysteinesulfinic acid In vitro. in Microorganisms
MDPI., 12(1), 34.
https://doi.org/10.3390/microorganisms12010034

Levent G, Božić A, Petrujkić B, Callaway TR, Poole TL, Crippen TL, Harvey RB, Ochoa-García P, Corral-Luna A, Yeater KM, Anderson RC. Assessment of potential anti-methanogenic and antimicrobial activity of ethyl nitroacetate, α-lipoic acid, taurine and L-cysteinesulfinic acid In vitro. in Microorganisms. 2024;12(1):34.
doi:10.3390/microorganisms12010034 .

Levent, Gizem, Božić, Aleksandar, Petrujkić, Branko, Callaway, Todd R., Poole, Toni L., Crippen, Tawni L., Harvey, Roger B., Ochoa-García, Pedro, Corral-Luna, Agustin, Yeater, Kathleen M., Anderson, Robin C., "Assessment of potential anti-methanogenic and antimicrobial activity of ethyl nitroacetate, α-lipoic acid, taurine and L-cysteinesulfinic acid In vitro" in Microorganisms, 12, no. 1 (2024):34,
https://doi.org/10.3390/microorganisms12010034 . .

TY  - JOUR
AU  - Levent, Gizem
AU  - Anderson, Robin C.
AU  - Petrujkić, Branko
AU  - Poole, Toni L.
AU  - He, Haiqi
AU  - Genovese, Kenneth J.
AU  - Hume, Michael E.
AU  - Beier, Ross C.
AU  - Harvey, Roger B.
AU  - Nisbet, David J.
PY  - 2021
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/2221
AB  - The gut of food-producing animals is a reservoir for foodborne pathogens. Thymol is bactericidal against foodborne pathogens but rapid absorption of thymol from the proximal gut precludes the delivery of effective concentrations to the lower gut where pathogens mainly colonize. Thymol-β-d-glucopyranoside is reported to be more resistant to absorption than thymol in everted jejunal segments and could potentially function as a prebiotic by resisting degradation and absorption in the proximal gut but being hydrolysable by microbial β-glycosidase in the distal gut. Previous in vitro studies showed bactericidal effects of thymol-β-d-glucopyranoside against Campylobacter, Escherichia coli, and Salmonella enterica serovar Typhimurium in the presence but not absence of intestinal microbes expressing β-glycosidase activity, indicating that hydrolysis was required to obtain antimicrobial activity. Presently, the oral administration of thymol-β-d-glucopyranoside was studied to examine the effects on intestinal carriage of Campylobacter, E. coli, and S. Typhimurium in swine. The effects of thymol-β-d-glucopyranoside or thymol on antimicrobial sensitivity of representative E. coli isolates and characterized Salmonella strains were also explored. Results from two in vivo studies revealed little antimicrobial effects of thymol-β-d-glucopyranoside on Campylobacter, E. coli, or S. Typhimurium in swine gut. These findings add credence to current thinking that hydrolysis and absorption of thymol-β-d-glucopyranoside and thymol may be sufficiently rapid within the proximal gut to preclude delivery to the distal gut. Antibiotic susceptibilities of selected bacterial isolates and strains were mainly unaffected by thymol. Further research is warranted to overcome obstacles, preventing the delivery of efficacious amounts of thymol-β-d-glucopyranoside to the lower gut.
PB  - MDPI
T2  - Microorganisms
T1  - Evaluation of Thymol-β-d-Glucopyranoside as a Potential Prebiotic Intervention to Reduce Carriage of Zoonotic Pathogens in Weaned and Feeder Pigs
VL  - 9
IS  - 4
SP  - 860
DO  - 10.3390/microorganisms9040860
ER  - 

@article{
author = "Levent, Gizem and Anderson, Robin C. and Petrujkić, Branko and Poole, Toni L. and He, Haiqi and Genovese, Kenneth J. and Hume, Michael E. and Beier, Ross C. and Harvey, Roger B. and Nisbet, David J.",
year = "2021",
abstract = "The gut of food-producing animals is a reservoir for foodborne pathogens. Thymol is bactericidal against foodborne pathogens but rapid absorption of thymol from the proximal gut precludes the delivery of effective concentrations to the lower gut where pathogens mainly colonize. Thymol-β-d-glucopyranoside is reported to be more resistant to absorption than thymol in everted jejunal segments and could potentially function as a prebiotic by resisting degradation and absorption in the proximal gut but being hydrolysable by microbial β-glycosidase in the distal gut. Previous in vitro studies showed bactericidal effects of thymol-β-d-glucopyranoside against Campylobacter, Escherichia coli, and Salmonella enterica serovar Typhimurium in the presence but not absence of intestinal microbes expressing β-glycosidase activity, indicating that hydrolysis was required to obtain antimicrobial activity. Presently, the oral administration of thymol-β-d-glucopyranoside was studied to examine the effects on intestinal carriage of Campylobacter, E. coli, and S. Typhimurium in swine. The effects of thymol-β-d-glucopyranoside or thymol on antimicrobial sensitivity of representative E. coli isolates and characterized Salmonella strains were also explored. Results from two in vivo studies revealed little antimicrobial effects of thymol-β-d-glucopyranoside on Campylobacter, E. coli, or S. Typhimurium in swine gut. These findings add credence to current thinking that hydrolysis and absorption of thymol-β-d-glucopyranoside and thymol may be sufficiently rapid within the proximal gut to preclude delivery to the distal gut. Antibiotic susceptibilities of selected bacterial isolates and strains were mainly unaffected by thymol. Further research is warranted to overcome obstacles, preventing the delivery of efficacious amounts of thymol-β-d-glucopyranoside to the lower gut.",
publisher = "MDPI",
journal = "Microorganisms",
title = "Evaluation of Thymol-β-d-Glucopyranoside as a Potential Prebiotic Intervention to Reduce Carriage of Zoonotic Pathogens in Weaned and Feeder Pigs",
volume = "9",
number = "4",
pages = "860",
doi = "10.3390/microorganisms9040860"
}

Levent, G., Anderson, R. C., Petrujkić, B., Poole, T. L., He, H., Genovese, K. J., Hume, M. E., Beier, R. C., Harvey, R. B.,& Nisbet, D. J.. (2021). Evaluation of Thymol-β-d-Glucopyranoside as a Potential Prebiotic Intervention to Reduce Carriage of Zoonotic Pathogens in Weaned and Feeder Pigs. in Microorganisms
MDPI., 9(4), 860.
https://doi.org/10.3390/microorganisms9040860

Levent G, Anderson RC, Petrujkić B, Poole TL, He H, Genovese KJ, Hume ME, Beier RC, Harvey RB, Nisbet DJ. Evaluation of Thymol-β-d-Glucopyranoside as a Potential Prebiotic Intervention to Reduce Carriage of Zoonotic Pathogens in Weaned and Feeder Pigs. in Microorganisms. 2021;9(4):860.
doi:10.3390/microorganisms9040860 .

Levent, Gizem, Anderson, Robin C., Petrujkić, Branko, Poole, Toni L., He, Haiqi, Genovese, Kenneth J., Hume, Michael E., Beier, Ross C., Harvey, Roger B., Nisbet, David J., "Evaluation of Thymol-β-d-Glucopyranoside as a Potential Prebiotic Intervention to Reduce Carriage of Zoonotic Pathogens in Weaned and Feeder Pigs" in Microorganisms, 9, no. 4 (2021):860,
https://doi.org/10.3390/microorganisms9040860 . .

TY  - JOUR
AU  - Anderson, Robin C.
AU  - Levent, Gizem
AU  - Petrujkić, Branko
AU  - Harvey, Roger B.
AU  - Hume, Michael E.
AU  - He, Haiqi
AU  - Genovese, Kenneth J.
AU  - Beier, Ross C.
AU  - Poole, Toni L.
AU  - Crippen, Tawni L.
AU  - Nisbet, David J.
PY  - 2021
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/2224
AB  - Strategies are sought to reduce the carriage and dissemination of zoonotic pathogens and antimicrobial-resistant microbes within food-producing animals and their production environment. Thymol (an essential oil) is a potent bactericide in vitro but in vivo efficacy has been inconsistent, largely due to its lipophilicity and absorption, which limits its passage and subsequent availability in the distal gastrointestinal tract. Conjugation of thymol to glucose to form thymol-β-d-glucopyranoside can decrease its absorption, but in vivo passage of effective concentrations to the lower gut remains suboptimal. Considering that contemporary swine diets often contain 5% or more added fat (to increase caloric density and reduce dustiness), we hypothesized that there may be sufficient residual fat in the distal intestinal tract to sequester free or conjugated thymol, thereby limiting the availability and subsequent effectiveness of this biocide. In support of this hypothesis, the anti-Salmonella Typhimurium effects of 6 mM free or conjugated thymol, expressed as log10-fold reductions of colony-forming units (CFU) ml−1, were diminished 90 and 58%, respectively, following 24-h in vitro anaerobic fecal incubation (at 39°C) with 3% added vegetable oil compared to reductions achieved during culture without added oil (6.1 log10 CFU ml−1). The antagonistic effect of vegetable oil and the bactericidal effect of free and conjugated thymol against Escherichia coli K88 tested similarly were diminished 86 and 84%, respectively, compared to reductions achieved in cultures incubated without added vegetable oil (5.7 log10 CFU ml−1). Inclusion of taurine (8 mg/ml), bile acids (0.6 mg/ml), or emulsifiers such as polyoxyethylene-40 stearate (0.2%), Tween 20, or Tween 80 (each at 1%) in the in vitro incubations had little effect on vegetable oil-caused inhibition of free or conjugated thymol. Based on these results, it seems reasonable to suspect that undigested lipid in the distal gut may limit the effectiveness of free or conjugated thymol. Accordingly, additional research is warranted to learn how to overcome obstacles diminishing bactericidal activity of free and conjugated thymol in the lower gastrointestinal tract of food-producing animals.
PB  - Frontiers Media
T2  - Frontiers in Veterinary Science
T1  - Antagonistic Effects of Lipids Against the Anti-Escherichia coli and Anti-Salmonella Activity of Thymol and Thymol-β-d-Glucopyranoside in Porcine Gut and Fecal Cultures In Vitro
VL  - 8
SP  - 1053
DO  - 10.3389/fvets.2021.751266
ER  - 

@article{
author = "Anderson, Robin C. and Levent, Gizem and Petrujkić, Branko and Harvey, Roger B. and Hume, Michael E. and He, Haiqi and Genovese, Kenneth J. and Beier, Ross C. and Poole, Toni L. and Crippen, Tawni L. and Nisbet, David J.",
year = "2021",
abstract = "Strategies are sought to reduce the carriage and dissemination of zoonotic pathogens and antimicrobial-resistant microbes within food-producing animals and their production environment. Thymol (an essential oil) is a potent bactericide in vitro but in vivo efficacy has been inconsistent, largely due to its lipophilicity and absorption, which limits its passage and subsequent availability in the distal gastrointestinal tract. Conjugation of thymol to glucose to form thymol-β-d-glucopyranoside can decrease its absorption, but in vivo passage of effective concentrations to the lower gut remains suboptimal. Considering that contemporary swine diets often contain 5% or more added fat (to increase caloric density and reduce dustiness), we hypothesized that there may be sufficient residual fat in the distal intestinal tract to sequester free or conjugated thymol, thereby limiting the availability and subsequent effectiveness of this biocide. In support of this hypothesis, the anti-Salmonella Typhimurium effects of 6 mM free or conjugated thymol, expressed as log10-fold reductions of colony-forming units (CFU) ml−1, were diminished 90 and 58%, respectively, following 24-h in vitro anaerobic fecal incubation (at 39°C) with 3% added vegetable oil compared to reductions achieved during culture without added oil (6.1 log10 CFU ml−1). The antagonistic effect of vegetable oil and the bactericidal effect of free and conjugated thymol against Escherichia coli K88 tested similarly were diminished 86 and 84%, respectively, compared to reductions achieved in cultures incubated without added vegetable oil (5.7 log10 CFU ml−1). Inclusion of taurine (8 mg/ml), bile acids (0.6 mg/ml), or emulsifiers such as polyoxyethylene-40 stearate (0.2%), Tween 20, or Tween 80 (each at 1%) in the in vitro incubations had little effect on vegetable oil-caused inhibition of free or conjugated thymol. Based on these results, it seems reasonable to suspect that undigested lipid in the distal gut may limit the effectiveness of free or conjugated thymol. Accordingly, additional research is warranted to learn how to overcome obstacles diminishing bactericidal activity of free and conjugated thymol in the lower gastrointestinal tract of food-producing animals.",
publisher = "Frontiers Media",
journal = "Frontiers in Veterinary Science",
title = "Antagonistic Effects of Lipids Against the Anti-Escherichia coli and Anti-Salmonella Activity of Thymol and Thymol-β-d-Glucopyranoside in Porcine Gut and Fecal Cultures In Vitro",
volume = "8",
pages = "1053",
doi = "10.3389/fvets.2021.751266"
}

Anderson, R. C., Levent, G., Petrujkić, B., Harvey, R. B., Hume, M. E., He, H., Genovese, K. J., Beier, R. C., Poole, T. L., Crippen, T. L.,& Nisbet, D. J.. (2021). Antagonistic Effects of Lipids Against the Anti-Escherichia coli and Anti-Salmonella Activity of Thymol and Thymol-β-d-Glucopyranoside in Porcine Gut and Fecal Cultures In Vitro. in Frontiers in Veterinary Science
Frontiers Media., 8, 1053.
https://doi.org/10.3389/fvets.2021.751266

Anderson RC, Levent G, Petrujkić B, Harvey RB, Hume ME, He H, Genovese KJ, Beier RC, Poole TL, Crippen TL, Nisbet DJ. Antagonistic Effects of Lipids Against the Anti-Escherichia coli and Anti-Salmonella Activity of Thymol and Thymol-β-d-Glucopyranoside in Porcine Gut and Fecal Cultures In Vitro. in Frontiers in Veterinary Science. 2021;8:1053.
doi:10.3389/fvets.2021.751266 .

Anderson, Robin C., Levent, Gizem, Petrujkić, Branko, Harvey, Roger B., Hume, Michael E., He, Haiqi, Genovese, Kenneth J., Beier, Ross C., Poole, Toni L., Crippen, Tawni L., Nisbet, David J., "Antagonistic Effects of Lipids Against the Anti-Escherichia coli and Anti-Salmonella Activity of Thymol and Thymol-β-d-Glucopyranoside in Porcine Gut and Fecal Cultures In Vitro" in Frontiers in Veterinary Science, 8 (2021):1053,
https://doi.org/10.3389/fvets.2021.751266 . .

TY  - JOUR
AU  - Petrujkić, Branko
AU  - Beier, Ross C.
AU  - He, Haiqi
AU  - Genovese, Kenneth J.
AU  - Swaggerty, Christina L.
AU  - Hume, Michael E.
AU  - Crippen, Tawni L.
AU  - Harvey, Roger B.
AU  - Anderson, Robin C.
AU  - Nisbet, David J.
PY  - 2018
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1627
AB  - BACKGROUNDNigella sativa L. (NS) is a plant containing bioactive constituents such as thymoquinone. Extracts of NS improve performance and reduce enteropathogen colonization in poultry and small ruminants, but studies with swine are lacking. In two different studies oral administration of NS extracts at doses equivalent to 0, 1.5 and 4.5g kg(-1) diet was assessed on piglet performance and intestinal carriage of wildtype Escherichia coli and Campylobacter, and Salmonella Typhimurium. RESULTSWildtype E. coli populations in the jejunal and rectal content collected 9days after treatment began were decreased (P0.05). Populations recovered from pigs treated with extract at 1.5 and 4.5g kg(-1) diet were 0.72-1.31 log(10) units lower than the controls (ranging from 6.05 to 6.61 log(10) CFU g(-1)). Wildtype Campylobacter and Salmonella Typhimurium were unaffected by NS treatment. Feed efficiency over the 9days improved linearly (P<0.05) from 3.88 with 0 NS-treated pigs to 1.47 and 1.41 with pigs treated with NS at 1.5 and 4.5g kg(-1) diet, respectively, possibly due to high glutamine/glutamic acid content of the NS extract. CONCLUSIONNS supplementation of weanling pigs improved feed efficiency and helped control intestinal E. coli during this vulnerable production phase.
PB  - Wiley, Hoboken
T2  - Journal of The Science of Food and Agriculture
T1  - Nigella sativa L. as an alternative antibiotic feed supplement and effect on growth performance in weanling pigs
VL  - 98
IS  - 8
SP  - 3175
EP  - 3181
DO  - 10.1002/jsfa.8823
ER  - 

@article{
author = "Petrujkić, Branko and Beier, Ross C. and He, Haiqi and Genovese, Kenneth J. and Swaggerty, Christina L. and Hume, Michael E. and Crippen, Tawni L. and Harvey, Roger B. and Anderson, Robin C. and Nisbet, David J.",
year = "2018",
abstract = "BACKGROUNDNigella sativa L. (NS) is a plant containing bioactive constituents such as thymoquinone. Extracts of NS improve performance and reduce enteropathogen colonization in poultry and small ruminants, but studies with swine are lacking. In two different studies oral administration of NS extracts at doses equivalent to 0, 1.5 and 4.5g kg(-1) diet was assessed on piglet performance and intestinal carriage of wildtype Escherichia coli and Campylobacter, and Salmonella Typhimurium. RESULTSWildtype E. coli populations in the jejunal and rectal content collected 9days after treatment began were decreased (P0.05). Populations recovered from pigs treated with extract at 1.5 and 4.5g kg(-1) diet were 0.72-1.31 log(10) units lower than the controls (ranging from 6.05 to 6.61 log(10) CFU g(-1)). Wildtype Campylobacter and Salmonella Typhimurium were unaffected by NS treatment. Feed efficiency over the 9days improved linearly (P<0.05) from 3.88 with 0 NS-treated pigs to 1.47 and 1.41 with pigs treated with NS at 1.5 and 4.5g kg(-1) diet, respectively, possibly due to high glutamine/glutamic acid content of the NS extract. CONCLUSIONNS supplementation of weanling pigs improved feed efficiency and helped control intestinal E. coli during this vulnerable production phase.",
publisher = "Wiley, Hoboken",
journal = "Journal of The Science of Food and Agriculture",
title = "Nigella sativa L. as an alternative antibiotic feed supplement and effect on growth performance in weanling pigs",
volume = "98",
number = "8",
pages = "3175-3181",
doi = "10.1002/jsfa.8823"
}

Petrujkić, B., Beier, R. C., He, H., Genovese, K. J., Swaggerty, C. L., Hume, M. E., Crippen, T. L., Harvey, R. B., Anderson, R. C.,& Nisbet, D. J.. (2018). Nigella sativa L. as an alternative antibiotic feed supplement and effect on growth performance in weanling pigs. in Journal of The Science of Food and Agriculture
Wiley, Hoboken., 98(8), 3175-3181.
https://doi.org/10.1002/jsfa.8823

Petrujkić B, Beier RC, He H, Genovese KJ, Swaggerty CL, Hume ME, Crippen TL, Harvey RB, Anderson RC, Nisbet DJ. Nigella sativa L. as an alternative antibiotic feed supplement and effect on growth performance in weanling pigs. in Journal of The Science of Food and Agriculture. 2018;98(8):3175-3181.
doi:10.1002/jsfa.8823 .

Petrujkić, Branko, Beier, Ross C., He, Haiqi, Genovese, Kenneth J., Swaggerty, Christina L., Hume, Michael E., Crippen, Tawni L., Harvey, Roger B., Anderson, Robin C., Nisbet, David J., "Nigella sativa L. as an alternative antibiotic feed supplement and effect on growth performance in weanling pigs" in Journal of The Science of Food and Agriculture, 98, no. 8 (2018):3175-3181,
https://doi.org/10.1002/jsfa.8823 . .

TY  - JOUR
AU  - Epps, Sharon V. R.
AU  - Harvey, Roger B.
AU  - Byrd, J. Allen
AU  - Petrujkić, Branko
AU  - Sedej, Ivana
AU  - Beier, Ross C.
AU  - Phillips, Timothy D.
AU  - Hume, Michael E.
AU  - Anderson, Robin C.
AU  - Nisbet, David J.
PY  - 2015
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1226
AB  - Campylobacter jejuni is an important human food-borne pathogen that can contaminate meat and poultry during processing. Consequently, strategies are sought to reduce the carriage of C. jejuni in food animals before they arrive at the abattoir. Thymol is a natural product that reduces survivability of Campylobacter in vitro, but its rapid absorption from the proximal alimentary tract limits its bactericidal efficacy in vivo. Thymol-beta-d-glucopyranoside is more resistant to absorption than free thymol, but its administration to chickens has not been reported. In the present studies, 1mM thymol-beta-d-glucopyranoside was shown to exhibit near equal anti-Campylobacter activity as 1mM thymol when incubated anaerobically in avian crop or cecal contents in vitro, resulting in reductions of 1.10-2.32 log(10) colony forming units mL(-1) in C. jejuni concentrations after 24h incubation. In a follow-up live animal study, oral administration of thymol-beta-d-glucopyranoside, but not free thymol, significantly lowered (>10-fold) recovery of Campylobacter from the crop of market-aged broilers when compared to placebo-treated controls (n = 6 broilers/treatment). Neither thymol-beta-d-glucopyranoside nor thymol affected recovery of Campylobacter from cecal contents of the treated broilers. These results indicate that rapid absorption or passage of free thymol from the crop precluded its anti-Campylobacter activity at this site and throughout the entire gastrointestinal tract. Conversely, lower recovery of Campylobacter from the crop of birds treated with thymol-beta-d-glucopyranoside indicates this conjugate was retained and able to be hydrolyzed to biologically active free thymol at this site as intended, yet was not sufficiently protected to allow passage of efficacious amounts of the intact glycoside to the lower gut. Nevertheless, these results warrant further research to see if higher doses or encapsulation of thymol-beta-d-glucopyranoside or similar glycosides may yield an efficacious additive to reduce carriage of Campylobacter as well as other pathogens throughout the avian gut.
PB  - Taylor & Francis Inc, Philadelphia
T2  - Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural
T1  - Comparative effect of thymol or its glucose conjugate, thymol-beta-d-glucopyranoside, on Campylobacter in avian gut contents
VL  - 50
IS  - 1
SP  - 55
EP  - 61
DO  - 10.1080/03601234.2015.965634
ER  - 

@article{
author = "Epps, Sharon V. R. and Harvey, Roger B. and Byrd, J. Allen and Petrujkić, Branko and Sedej, Ivana and Beier, Ross C. and Phillips, Timothy D. and Hume, Michael E. and Anderson, Robin C. and Nisbet, David J.",
year = "2015",
abstract = "Campylobacter jejuni is an important human food-borne pathogen that can contaminate meat and poultry during processing. Consequently, strategies are sought to reduce the carriage of C. jejuni in food animals before they arrive at the abattoir. Thymol is a natural product that reduces survivability of Campylobacter in vitro, but its rapid absorption from the proximal alimentary tract limits its bactericidal efficacy in vivo. Thymol-beta-d-glucopyranoside is more resistant to absorption than free thymol, but its administration to chickens has not been reported. In the present studies, 1mM thymol-beta-d-glucopyranoside was shown to exhibit near equal anti-Campylobacter activity as 1mM thymol when incubated anaerobically in avian crop or cecal contents in vitro, resulting in reductions of 1.10-2.32 log(10) colony forming units mL(-1) in C. jejuni concentrations after 24h incubation. In a follow-up live animal study, oral administration of thymol-beta-d-glucopyranoside, but not free thymol, significantly lowered (>10-fold) recovery of Campylobacter from the crop of market-aged broilers when compared to placebo-treated controls (n = 6 broilers/treatment). Neither thymol-beta-d-glucopyranoside nor thymol affected recovery of Campylobacter from cecal contents of the treated broilers. These results indicate that rapid absorption or passage of free thymol from the crop precluded its anti-Campylobacter activity at this site and throughout the entire gastrointestinal tract. Conversely, lower recovery of Campylobacter from the crop of birds treated with thymol-beta-d-glucopyranoside indicates this conjugate was retained and able to be hydrolyzed to biologically active free thymol at this site as intended, yet was not sufficiently protected to allow passage of efficacious amounts of the intact glycoside to the lower gut. Nevertheless, these results warrant further research to see if higher doses or encapsulation of thymol-beta-d-glucopyranoside or similar glycosides may yield an efficacious additive to reduce carriage of Campylobacter as well as other pathogens throughout the avian gut.",
publisher = "Taylor & Francis Inc, Philadelphia",
journal = "Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural",
title = "Comparative effect of thymol or its glucose conjugate, thymol-beta-d-glucopyranoside, on Campylobacter in avian gut contents",
volume = "50",
number = "1",
pages = "55-61",
doi = "10.1080/03601234.2015.965634"
}

Epps, S. V. R., Harvey, R. B., Byrd, J. A., Petrujkić, B., Sedej, I., Beier, R. C., Phillips, T. D., Hume, M. E., Anderson, R. C.,& Nisbet, D. J.. (2015). Comparative effect of thymol or its glucose conjugate, thymol-beta-d-glucopyranoside, on Campylobacter in avian gut contents. in Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural
Taylor & Francis Inc, Philadelphia., 50(1), 55-61.
https://doi.org/10.1080/03601234.2015.965634

Epps SVR, Harvey RB, Byrd JA, Petrujkić B, Sedej I, Beier RC, Phillips TD, Hume ME, Anderson RC, Nisbet DJ. Comparative effect of thymol or its glucose conjugate, thymol-beta-d-glucopyranoside, on Campylobacter in avian gut contents. in Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural. 2015;50(1):55-61.
doi:10.1080/03601234.2015.965634 .

Epps, Sharon V. R., Harvey, Roger B., Byrd, J. Allen, Petrujkić, Branko, Sedej, Ivana, Beier, Ross C., Phillips, Timothy D., Hume, Michael E., Anderson, Robin C., Nisbet, David J., "Comparative effect of thymol or its glucose conjugate, thymol-beta-d-glucopyranoside, on Campylobacter in avian gut contents" in Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural, 50, no. 1 (2015):55-61,
https://doi.org/10.1080/03601234.2015.965634 . .

TY  - JOUR
AU  - Epps, Sharon V. R.
AU  - Petrujkić, Branko
AU  - Sedej, Ivana
AU  - Krueger, Nathan A.
AU  - Harvey, Roger B.
AU  - Beier, Ross C.
AU  - Stanton, Thad B.
AU  - Phillips, Timothy D.
AU  - Anderson, Robin C.
AU  - Nisbet, David J.
PY  - 2015
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1218
AB  - Thymol is a natural product that exhibits antimicrobial activity in vitro but in vivo results indicate that absorption within the proximal alimentary tract precludes its delivery to the distal gut. Presently, the anti-Campylobacter activity of thymol was compared against that of thymol-beta-D-glucopyranoside, the latter being resistant to absorption. When treated with 1 mM thymol, Campylobacter coli and jejuni were reduced during pure or co-culture with a beta-glycoside-hydrolysing Parabacteroides distasonis. Thymol-beta-D-glucopyranoside treatment (1 mM) did not reduce C coli and jejuni during pure culture but did during co-culture with P. distasonis or during mixed culture with porcine or bovine faecal microbes possessing beta-glycoside-hydrolysing activity. Fermentation acid production was reduced by thymol-beta-D-glucopyranoside treatment, indicating that fermentation was inhibited, which may limit its application to just before harvest. Results suggest that thymol-beta-D-glucopyranoside or similar beta-glycosides may be able to escape absorption within the proximal gut and become activated by bacterial beta-glycosidases in the distal gut. Published by Elsevier Ltd.
PB  - Elsevier Sci Ltd, Oxford
T2  - Food Chemistry
T1  - Comparison of anti-Campylobacter activity of free thymol and thymol-beta-D-glucopyranoside in absence or presence of beta-glycoside-hydrolysing gut bacteria
VL  - 173
SP  - 92
EP  - 98
DO  - 10.1016/j.foodchem.2014.10.007
ER  - 

@article{
author = "Epps, Sharon V. R. and Petrujkić, Branko and Sedej, Ivana and Krueger, Nathan A. and Harvey, Roger B. and Beier, Ross C. and Stanton, Thad B. and Phillips, Timothy D. and Anderson, Robin C. and Nisbet, David J.",
year = "2015",
abstract = "Thymol is a natural product that exhibits antimicrobial activity in vitro but in vivo results indicate that absorption within the proximal alimentary tract precludes its delivery to the distal gut. Presently, the anti-Campylobacter activity of thymol was compared against that of thymol-beta-D-glucopyranoside, the latter being resistant to absorption. When treated with 1 mM thymol, Campylobacter coli and jejuni were reduced during pure or co-culture with a beta-glycoside-hydrolysing Parabacteroides distasonis. Thymol-beta-D-glucopyranoside treatment (1 mM) did not reduce C coli and jejuni during pure culture but did during co-culture with P. distasonis or during mixed culture with porcine or bovine faecal microbes possessing beta-glycoside-hydrolysing activity. Fermentation acid production was reduced by thymol-beta-D-glucopyranoside treatment, indicating that fermentation was inhibited, which may limit its application to just before harvest. Results suggest that thymol-beta-D-glucopyranoside or similar beta-glycosides may be able to escape absorption within the proximal gut and become activated by bacterial beta-glycosidases in the distal gut. Published by Elsevier Ltd.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Food Chemistry",
title = "Comparison of anti-Campylobacter activity of free thymol and thymol-beta-D-glucopyranoside in absence or presence of beta-glycoside-hydrolysing gut bacteria",
volume = "173",
pages = "92-98",
doi = "10.1016/j.foodchem.2014.10.007"
}

Epps, S. V. R., Petrujkić, B., Sedej, I., Krueger, N. A., Harvey, R. B., Beier, R. C., Stanton, T. B., Phillips, T. D., Anderson, R. C.,& Nisbet, D. J.. (2015). Comparison of anti-Campylobacter activity of free thymol and thymol-beta-D-glucopyranoside in absence or presence of beta-glycoside-hydrolysing gut bacteria. in Food Chemistry
Elsevier Sci Ltd, Oxford., 173, 92-98.
https://doi.org/10.1016/j.foodchem.2014.10.007

Epps SVR, Petrujkić B, Sedej I, Krueger NA, Harvey RB, Beier RC, Stanton TB, Phillips TD, Anderson RC, Nisbet DJ. Comparison of anti-Campylobacter activity of free thymol and thymol-beta-D-glucopyranoside in absence or presence of beta-glycoside-hydrolysing gut bacteria. in Food Chemistry. 2015;173:92-98.
doi:10.1016/j.foodchem.2014.10.007 .

Epps, Sharon V. R., Petrujkić, Branko, Sedej, Ivana, Krueger, Nathan A., Harvey, Roger B., Beier, Ross C., Stanton, Thad B., Phillips, Timothy D., Anderson, Robin C., Nisbet, David J., "Comparison of anti-Campylobacter activity of free thymol and thymol-beta-D-glucopyranoside in absence or presence of beta-glycoside-hydrolysing gut bacteria" in Food Chemistry, 173 (2015):92-98,
https://doi.org/10.1016/j.foodchem.2014.10.007 . .

TY  - JOUR
AU  - Petrujkić, Branko
AU  - Šefer, Dragan
AU  - Jovanović, Ivan B.
AU  - Jovičin, Milovan
AU  - Janković, S.
AU  - Jakovljević, Goran
AU  - Beier, Ross C.
AU  - Anderson, Robin C.
PY  - 2014
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1089
AB  - The aim of this study was to determine how dietary supplementation of inorganic and organic selenium affects the selenium concentration and glutathione peroxidase activity in blood and sperm of sexually mature stud boars. Twenty-four boars of the Large White, Landrace, Pietrain and Duroc breeds of optimal breeding age (on average 2.5 years old) were used. The study lasted 90 days. The boars were randomly assigned to one of three dietary treatment groups: T1 = control; no added selenium (n = 8 boars), T2 = added 0.3 ppm inorganic selenium (sodium selenite, Microgran (R) Se 1% BMP) (n = 8 boars), and T3 = added 0.3 ppm organic selenium (Se-yeast, Sel-Plex 2000 (R)) (n = 8 boars). The concentration of selenium was determined in whole blood and semen, while the activity of glutathione peroxidase (GPx) was measured in blood plasma and semen. In order to measure GPx activity in semen, reactivation of the enzymatic GPx activity was performed. The determined selenium concentration in blood was lowest in the non-supplemented group of boars. Blood plasma GPx activity was higher in boars fed organic selenium than in boars fed a diet without supplemented selenium. While the supplementation of sodium selenite significantly increased GPx activity in boar semen. The highest-concentration of selenium in semen at the end of the trial was determined in the group of boars supplemented with organic selenium, somewhat lower in boars fed supplemented inorganic selenium, and the lowest in the non-supplemented group of boars. The only significant difference between the selenite and Se-yeast diet supplementation was observed in the Se concentration of the semen. The supplementation of selenium affected semen quality, and organic selenium improved the progressive motility of the spermatozoa and increased their resistance in hypo-osmotic and thermal tests. The storage ability of short term preserved semen was improved by organic selenium supplementation, as well as also increasing the fertility rate in gilts.
PB  - Elsevier Science Bv, Amsterdam
T2  - Animal Feed Science and Technology
T1  - Effects of commercial selenium products on glutathione peroxidase activity and semen quality in stud boars
VL  - 197
SP  - 194
EP  - 205
DO  - 10.1016/j.anifeedsci.2014.09.001
ER  - 

@article{
author = "Petrujkić, Branko and Šefer, Dragan and Jovanović, Ivan B. and Jovičin, Milovan and Janković, S. and Jakovljević, Goran and Beier, Ross C. and Anderson, Robin C.",
year = "2014",
abstract = "The aim of this study was to determine how dietary supplementation of inorganic and organic selenium affects the selenium concentration and glutathione peroxidase activity in blood and sperm of sexually mature stud boars. Twenty-four boars of the Large White, Landrace, Pietrain and Duroc breeds of optimal breeding age (on average 2.5 years old) were used. The study lasted 90 days. The boars were randomly assigned to one of three dietary treatment groups: T1 = control; no added selenium (n = 8 boars), T2 = added 0.3 ppm inorganic selenium (sodium selenite, Microgran (R) Se 1% BMP) (n = 8 boars), and T3 = added 0.3 ppm organic selenium (Se-yeast, Sel-Plex 2000 (R)) (n = 8 boars). The concentration of selenium was determined in whole blood and semen, while the activity of glutathione peroxidase (GPx) was measured in blood plasma and semen. In order to measure GPx activity in semen, reactivation of the enzymatic GPx activity was performed. The determined selenium concentration in blood was lowest in the non-supplemented group of boars. Blood plasma GPx activity was higher in boars fed organic selenium than in boars fed a diet without supplemented selenium. While the supplementation of sodium selenite significantly increased GPx activity in boar semen. The highest-concentration of selenium in semen at the end of the trial was determined in the group of boars supplemented with organic selenium, somewhat lower in boars fed supplemented inorganic selenium, and the lowest in the non-supplemented group of boars. The only significant difference between the selenite and Se-yeast diet supplementation was observed in the Se concentration of the semen. The supplementation of selenium affected semen quality, and organic selenium improved the progressive motility of the spermatozoa and increased their resistance in hypo-osmotic and thermal tests. The storage ability of short term preserved semen was improved by organic selenium supplementation, as well as also increasing the fertility rate in gilts.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Animal Feed Science and Technology",
title = "Effects of commercial selenium products on glutathione peroxidase activity and semen quality in stud boars",
volume = "197",
pages = "194-205",
doi = "10.1016/j.anifeedsci.2014.09.001"
}

Petrujkić, B., Šefer, D., Jovanović, I. B., Jovičin, M., Janković, S., Jakovljević, G., Beier, R. C.,& Anderson, R. C.. (2014). Effects of commercial selenium products on glutathione peroxidase activity and semen quality in stud boars. in Animal Feed Science and Technology
Elsevier Science Bv, Amsterdam., 197, 194-205.
https://doi.org/10.1016/j.anifeedsci.2014.09.001

Petrujkić B, Šefer D, Jovanović IB, Jovičin M, Janković S, Jakovljević G, Beier RC, Anderson RC. Effects of commercial selenium products on glutathione peroxidase activity and semen quality in stud boars. in Animal Feed Science and Technology. 2014;197:194-205.
doi:10.1016/j.anifeedsci.2014.09.001 .

Petrujkić, Branko, Šefer, Dragan, Jovanović, Ivan B., Jovičin, Milovan, Janković, S., Jakovljević, Goran, Beier, Ross C., Anderson, Robin C., "Effects of commercial selenium products on glutathione peroxidase activity and semen quality in stud boars" in Animal Feed Science and Technology, 197 (2014):194-205,
https://doi.org/10.1016/j.anifeedsci.2014.09.001 . .

TY  - JOUR
AU  - Petrujkić, Branko
AU  - Sedej, Ivana
AU  - Beier, Ross C.
AU  - Anderson, Robin C.
AU  - Harvey, Roger B.
AU  - Epps, Sharon V. R.
AU  - Stipanović, Robert D.
AU  - Krueger, Nathan A.
AU  - Nisbet, David J.
PY  - 2013
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1016
AB  - Food-producing animals are reservoirs of Campylobacter, a leading bacterial cause of human foodborne illness. The natural product thymol can reduce the survivability of Campylobacter, but its rapid absorption in the proximal gastrointestinal tract may preclude its use as a feed additive to reduce intestinal colonization of these pathogens. This work examined the ex vivo absorption of thymol and thymol-beta-D-glucopyranoside in everted porcine jejunal segments, as the latter was hypothesized to be more resistant to absorption. A modified gas chromatography and extraction method was developed to determine 1.0-500 mg/L thymol. From 1 and 3 mM solutions, 0.293 +/- 0.04 and 0.898 +/- 0.212 mM thymol, respectively, p = 0.0347, were absorbed, and 0.125 +/- 0.041 and 0.317 +/- 0.143 mM thymol-beta-D-glucopyranoside, respectively, p = 0.0892, were absorbed. Results indicate that thymol-beta-D-glucopyranoside was absorbed 2.3 to 2.8 times less effectively than thymol, thus providing evidence that thymol-beta-D-glucopyranoside may potentially be used as a feed additive to transport thymol to the piglet lower gut.
PB  - Amer Chemical Soc, Washington
T2  - Journal of Agricultural and Food Chemistry
T1  - Ex Vivo Absorption of Thymol and Thymol-beta-D-glucopyranoside in Piglet Everted Jejunal Segments
VL  - 61
IS  - 15
SP  - 3757
EP  - 3762
DO  - 10.1021/jf401013a
ER  - 

@article{
author = "Petrujkić, Branko and Sedej, Ivana and Beier, Ross C. and Anderson, Robin C. and Harvey, Roger B. and Epps, Sharon V. R. and Stipanović, Robert D. and Krueger, Nathan A. and Nisbet, David J.",
year = "2013",
abstract = "Food-producing animals are reservoirs of Campylobacter, a leading bacterial cause of human foodborne illness. The natural product thymol can reduce the survivability of Campylobacter, but its rapid absorption in the proximal gastrointestinal tract may preclude its use as a feed additive to reduce intestinal colonization of these pathogens. This work examined the ex vivo absorption of thymol and thymol-beta-D-glucopyranoside in everted porcine jejunal segments, as the latter was hypothesized to be more resistant to absorption. A modified gas chromatography and extraction method was developed to determine 1.0-500 mg/L thymol. From 1 and 3 mM solutions, 0.293 +/- 0.04 and 0.898 +/- 0.212 mM thymol, respectively, p = 0.0347, were absorbed, and 0.125 +/- 0.041 and 0.317 +/- 0.143 mM thymol-beta-D-glucopyranoside, respectively, p = 0.0892, were absorbed. Results indicate that thymol-beta-D-glucopyranoside was absorbed 2.3 to 2.8 times less effectively than thymol, thus providing evidence that thymol-beta-D-glucopyranoside may potentially be used as a feed additive to transport thymol to the piglet lower gut.",
publisher = "Amer Chemical Soc, Washington",
journal = "Journal of Agricultural and Food Chemistry",
title = "Ex Vivo Absorption of Thymol and Thymol-beta-D-glucopyranoside in Piglet Everted Jejunal Segments",
volume = "61",
number = "15",
pages = "3757-3762",
doi = "10.1021/jf401013a"
}

Petrujkić, B., Sedej, I., Beier, R. C., Anderson, R. C., Harvey, R. B., Epps, S. V. R., Stipanović, R. D., Krueger, N. A.,& Nisbet, D. J.. (2013). Ex Vivo Absorption of Thymol and Thymol-beta-D-glucopyranoside in Piglet Everted Jejunal Segments. in Journal of Agricultural and Food Chemistry
Amer Chemical Soc, Washington., 61(15), 3757-3762.
https://doi.org/10.1021/jf401013a

Petrujkić B, Sedej I, Beier RC, Anderson RC, Harvey RB, Epps SVR, Stipanović RD, Krueger NA, Nisbet DJ. Ex Vivo Absorption of Thymol and Thymol-beta-D-glucopyranoside in Piglet Everted Jejunal Segments. in Journal of Agricultural and Food Chemistry. 2013;61(15):3757-3762.
doi:10.1021/jf401013a .

Petrujkić, Branko, Sedej, Ivana, Beier, Ross C., Anderson, Robin C., Harvey, Roger B., Epps, Sharon V. R., Stipanović, Robert D., Krueger, Nathan A., Nisbet, David J., "Ex Vivo Absorption of Thymol and Thymol-beta-D-glucopyranoside in Piglet Everted Jejunal Segments" in Journal of Agricultural and Food Chemistry, 61, no. 15 (2013):3757-3762,
https://doi.org/10.1021/jf401013a . .

TY  - CONF
AU  - Beier, Ross C.
AU  - Petrujkić, Branko
AU  - Sedej, Ivana
AU  - Anderson, Robin C.
AU  - Nedeljković-Trailović, Jelena
AU  - Nisbet, David J.
PY  - 2012
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/896
AB  - Aromatic plants and their extracts have been used in human and veterinary medicine since ancient times. However, the exact mode of action and kinetics or these compounds are generally poorly understood. The aim of this work was to determine the degradation and absorption kinetics of thymol in the pig stomach. An improved GC method was developed to easily measure the thymol content in samples extracted into ethylacetate. The GC separation was achieved on a 30 m × 0.25 mm ™ 0.25 mm film thickness Equity™ - 1701 fused silica capillary column, resulting in a standard curve over 1-500 mg/L. Isolated pig stomach was incubated for 360 min at 37 °C in an incubator with 95% O2:5% CO2 atmosphere on an oscillating plate at 40 rpm. Tyrode's solution pH 6.5 was used as incubation fluid. Pig stomach contents were inoculated with thymol (1.7 mM), and samples were collected at intervals during incubation and were extracted as is or after treatment with a lysis buffer. The lysis buffer was used to release thymol that may have been internalized by endogenous bacteria. Thymol concentrations in unlysed stomach samples decreased rapidly, being reduced more than twofold during 30 min incubations. Conversely, thymol concentrations in lysed samples increased rapidly, following 60 min incubations, concentrations were achieved at twice those measured in unlysed stomach contents at the beginning of the incubation. Thymol concentrations in lysed samples remained nearly double those in unlysed samples even after 360 min. Differing thymol concentrations measured in lysed and unlysed samples suggest luminal depletion resulting from bacterial uptake of thymol. Upon ingestion, luminal depletion would be expected to occur rapidly in stomach contents, but whether this sequestration would make thymol unavailable for absorption in the stomach or more distally is not known. Further investigation is needed to identify the bacteria responsible for this "thymol sequestration" phenomenon.
PB  - University of Novi Sad, Faculty of Technology
C3  - CEFood 2012 - Proceedings of 6th Central European Congress on Food
T1  - Thymol depletion in the pig stomach
SP  - 279
EP  - 283
UR  - https://hdl.handle.net/21.15107/rcub_veterinar_896
ER  - 

@conference{
author = "Beier, Ross C. and Petrujkić, Branko and Sedej, Ivana and Anderson, Robin C. and Nedeljković-Trailović, Jelena and Nisbet, David J.",
year = "2012",
abstract = "Aromatic plants and their extracts have been used in human and veterinary medicine since ancient times. However, the exact mode of action and kinetics or these compounds are generally poorly understood. The aim of this work was to determine the degradation and absorption kinetics of thymol in the pig stomach. An improved GC method was developed to easily measure the thymol content in samples extracted into ethylacetate. The GC separation was achieved on a 30 m × 0.25 mm ™ 0.25 mm film thickness Equity™ - 1701 fused silica capillary column, resulting in a standard curve over 1-500 mg/L. Isolated pig stomach was incubated for 360 min at 37 °C in an incubator with 95% O2:5% CO2 atmosphere on an oscillating plate at 40 rpm. Tyrode's solution pH 6.5 was used as incubation fluid. Pig stomach contents were inoculated with thymol (1.7 mM), and samples were collected at intervals during incubation and were extracted as is or after treatment with a lysis buffer. The lysis buffer was used to release thymol that may have been internalized by endogenous bacteria. Thymol concentrations in unlysed stomach samples decreased rapidly, being reduced more than twofold during 30 min incubations. Conversely, thymol concentrations in lysed samples increased rapidly, following 60 min incubations, concentrations were achieved at twice those measured in unlysed stomach contents at the beginning of the incubation. Thymol concentrations in lysed samples remained nearly double those in unlysed samples even after 360 min. Differing thymol concentrations measured in lysed and unlysed samples suggest luminal depletion resulting from bacterial uptake of thymol. Upon ingestion, luminal depletion would be expected to occur rapidly in stomach contents, but whether this sequestration would make thymol unavailable for absorption in the stomach or more distally is not known. Further investigation is needed to identify the bacteria responsible for this "thymol sequestration" phenomenon.",
publisher = "University of Novi Sad, Faculty of Technology",
journal = "CEFood 2012 - Proceedings of 6th Central European Congress on Food",
title = "Thymol depletion in the pig stomach",
pages = "279-283",
url = "https://hdl.handle.net/21.15107/rcub_veterinar_896"
}

Beier, R. C., Petrujkić, B., Sedej, I., Anderson, R. C., Nedeljković-Trailović, J.,& Nisbet, D. J.. (2012). Thymol depletion in the pig stomach. in CEFood 2012 - Proceedings of 6th Central European Congress on Food
University of Novi Sad, Faculty of Technology., 279-283.
https://hdl.handle.net/21.15107/rcub_veterinar_896

Beier RC, Petrujkić B, Sedej I, Anderson RC, Nedeljković-Trailović J, Nisbet DJ. Thymol depletion in the pig stomach. in CEFood 2012 - Proceedings of 6th Central European Congress on Food. 2012;:279-283.
https://hdl.handle.net/21.15107/rcub_veterinar_896 .

Beier, Ross C., Petrujkić, Branko, Sedej, Ivana, Anderson, Robin C., Nedeljković-Trailović, Jelena, Nisbet, David J., "Thymol depletion in the pig stomach" in CEFood 2012 - Proceedings of 6th Central European Congress on Food (2012):279-283,
https://hdl.handle.net/21.15107/rcub_veterinar_896 .

TY  - CONF
AU  - Sedej, Ivana
AU  - Petrujkić, Branko
AU  - Beier, Ross C.
AU  - Anderson, Robin C.
AU  - Nisbet, David J.
PY  - 2012
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/940
PB  - American Chemical Society
C3  - Abstracts of Papers of The American Chemical Society
T1  - Gas chromatography determination of thymol in swine in vitro everted gut segments treated with a conjugate of thymol
VL  - 243
UR  - https://hdl.handle.net/21.15107/rcub_veterinar_940
ER  - 

@conference{
author = "Sedej, Ivana and Petrujkić, Branko and Beier, Ross C. and Anderson, Robin C. and Nisbet, David J.",
year = "2012",
publisher = "American Chemical Society",
journal = "Abstracts of Papers of The American Chemical Society",
title = "Gas chromatography determination of thymol in swine in vitro everted gut segments treated with a conjugate of thymol",
volume = "243",
url = "https://hdl.handle.net/21.15107/rcub_veterinar_940"
}

Sedej, I., Petrujkić, B., Beier, R. C., Anderson, R. C.,& Nisbet, D. J.. (2012). Gas chromatography determination of thymol in swine in vitro everted gut segments treated with a conjugate of thymol. in Abstracts of Papers of The American Chemical Society
American Chemical Society., 243.
https://hdl.handle.net/21.15107/rcub_veterinar_940

Sedej I, Petrujkić B, Beier RC, Anderson RC, Nisbet DJ. Gas chromatography determination of thymol in swine in vitro everted gut segments treated with a conjugate of thymol. in Abstracts of Papers of The American Chemical Society. 2012;243.
https://hdl.handle.net/21.15107/rcub_veterinar_940 .

Sedej, Ivana, Petrujkić, Branko, Beier, Ross C., Anderson, Robin C., Nisbet, David J., "Gas chromatography determination of thymol in swine in vitro everted gut segments treated with a conjugate of thymol" in Abstracts of Papers of The American Chemical Society, 243 (2012),
https://hdl.handle.net/21.15107/rcub_veterinar_940 .

TY  - JOUR
AU  - Dimitrijević, Mirjana
AU  - Anderson, Robin C.
AU  - Karabasil, Nedjeljko
AU  - Pavlićević, Nataša
AU  - Jovanović, S.
AU  - Nedeljković-Trailović, Jelena
AU  - Teodorović, Vlado
AU  - Marković, Maja
AU  - Dojčinović, Slobodan
PY  - 2011
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/812
AB  - The presence of Listeria monocytogenes on the surfaces of equipment and worker's hands during different production stages, as well as on fish skin and meat during processing and storage of cold-smoked trout, was investigated. Listeria monocytogenes was recovered from 10 (6.06%) of a total 165 cotton-swabbed samples collected from the surfaces of equipment and worker's hands at two separate processing facilities. Of 105 samples collected from fish skin and meat during various production steps in both processing plants, 14 (13.33%) were confirmed culture-positive for L. monocytogenes, with recovery being most frequently in samples collected in the area before vacuum packaging. Recovery rates at two different Serbian processing plants did not differ (p lt 0.05), but suggested that different L. monocytogenes serotypes appeared to be resident within each processing plant and may have contributed to the final product contamination. From all smoked trout samples collected during 7, 14, 21 and 28 days of storage at 4ºC only two were culture-positive for L. monocytogenes serotype 1/2a and both of these were collected on the 7th day of storage. Conversely, 4, 3 and 1 sample(s) were contaminated with L. monocytogenes serotypes 1/2a and 1/2b after 7, 14 and 21 days storage at 10ºC. Listeria monocytogenes was not recovered from smoked trout stored 28 days at 10ºC. Results emphasize the importance of adhering to strict hygienic and quality control standards throughout the processing environment.
AB  - U ovom radu je ispitivan nalaz Listeria monocytogenes na površini opreme, rukama radnika, kao i koži i mesu ribe tokom različitih faza proizvodnje hladno dimljene pastrmke. Listeria monocytogenes je utvrđena kod 10 (6,06%) od ukupno 165 briseva sa površine opreme i ruku radnika iz dva proizvodna pogona. Od 105 uzoraka mesa ribe sa kožom, uzetih tokom proizvodnje u oba proizvodna pogona, 14 (13,33%) je bilo pozitivno na L. monocytogenes, pri čemu je nalaz i preživljavanje ove bakterije bio najčešći u uzorcima uzetih tokom proizvodnih procesa pre vakuum pakovanja. Stepen preživljavanja bakterije se nije značajno razlikovao u dva proizvodna pogona (P>0.05), ali se može zaključiti da su pojedini serotipovi L. monocytogenes otporniji unutar pogona i mogu dovesti do kontaminacije finalnog proizvoda. Od svih uzoraka iz oba proizvodna pogona, skladištenih tokom 7, 14, 21 i 28 dana pri temperaturi od 4ºC, dva su uzorka, sedmog dana skladištenja, bila kontaminirana L. monocytogenes serotipom 1/2a. Nasuprot tome, pri temperaturi od 10ºC, nalaz L. monocytogenes serotipa 1/2a i 1/2b ustanovljen je kod 4 uzorka sedmog dana skladištenja, 3 uzorka četrnaestog dana i jednog uzorka tokom dvadesetprvog dana skladištenja. Listeria monocytogenes nije preživela u uzorcima nakon skladištenja tokom 28 dana. Rezultati ukazuju na značaj primene strogih higijenskih mera, kao i standarda kontrole kvaliteta tokom proizvodnje hladno dimljene ribe.
PB  - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
T2  - Acta Veterinaria-Beograd
T1  - Environmental prevalence and persistence of Listeria monocytogenes in cold-smoked trout processing plants
T1  - Nalaz i preživljavanje Listeria monocytogenes u pogonima za proizvodnju hladno dimljene pastrmke
VL  - 61
IS  - 4
SP  - 429
EP  - 442
DO  - 10.2298/AVB1104429D
ER  - 

@article{
author = "Dimitrijević, Mirjana and Anderson, Robin C. and Karabasil, Nedjeljko and Pavlićević, Nataša and Jovanović, S. and Nedeljković-Trailović, Jelena and Teodorović, Vlado and Marković, Maja and Dojčinović, Slobodan",
year = "2011",
abstract = "The presence of Listeria monocytogenes on the surfaces of equipment and worker's hands during different production stages, as well as on fish skin and meat during processing and storage of cold-smoked trout, was investigated. Listeria monocytogenes was recovered from 10 (6.06%) of a total 165 cotton-swabbed samples collected from the surfaces of equipment and worker's hands at two separate processing facilities. Of 105 samples collected from fish skin and meat during various production steps in both processing plants, 14 (13.33%) were confirmed culture-positive for L. monocytogenes, with recovery being most frequently in samples collected in the area before vacuum packaging. Recovery rates at two different Serbian processing plants did not differ (p lt 0.05), but suggested that different L. monocytogenes serotypes appeared to be resident within each processing plant and may have contributed to the final product contamination. From all smoked trout samples collected during 7, 14, 21 and 28 days of storage at 4ºC only two were culture-positive for L. monocytogenes serotype 1/2a and both of these were collected on the 7th day of storage. Conversely, 4, 3 and 1 sample(s) were contaminated with L. monocytogenes serotypes 1/2a and 1/2b after 7, 14 and 21 days storage at 10ºC. Listeria monocytogenes was not recovered from smoked trout stored 28 days at 10ºC. Results emphasize the importance of adhering to strict hygienic and quality control standards throughout the processing environment., U ovom radu je ispitivan nalaz Listeria monocytogenes na površini opreme, rukama radnika, kao i koži i mesu ribe tokom različitih faza proizvodnje hladno dimljene pastrmke. Listeria monocytogenes je utvrđena kod 10 (6,06%) od ukupno 165 briseva sa površine opreme i ruku radnika iz dva proizvodna pogona. Od 105 uzoraka mesa ribe sa kožom, uzetih tokom proizvodnje u oba proizvodna pogona, 14 (13,33%) je bilo pozitivno na L. monocytogenes, pri čemu je nalaz i preživljavanje ove bakterije bio najčešći u uzorcima uzetih tokom proizvodnih procesa pre vakuum pakovanja. Stepen preživljavanja bakterije se nije značajno razlikovao u dva proizvodna pogona (P>0.05), ali se može zaključiti da su pojedini serotipovi L. monocytogenes otporniji unutar pogona i mogu dovesti do kontaminacije finalnog proizvoda. Od svih uzoraka iz oba proizvodna pogona, skladištenih tokom 7, 14, 21 i 28 dana pri temperaturi od 4ºC, dva su uzorka, sedmog dana skladištenja, bila kontaminirana L. monocytogenes serotipom 1/2a. Nasuprot tome, pri temperaturi od 10ºC, nalaz L. monocytogenes serotipa 1/2a i 1/2b ustanovljen je kod 4 uzorka sedmog dana skladištenja, 3 uzorka četrnaestog dana i jednog uzorka tokom dvadesetprvog dana skladištenja. Listeria monocytogenes nije preživela u uzorcima nakon skladištenja tokom 28 dana. Rezultati ukazuju na značaj primene strogih higijenskih mera, kao i standarda kontrole kvaliteta tokom proizvodnje hladno dimljene ribe.",
publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd",
journal = "Acta Veterinaria-Beograd",
title = "Environmental prevalence and persistence of Listeria monocytogenes in cold-smoked trout processing plants, Nalaz i preživljavanje Listeria monocytogenes u pogonima za proizvodnju hladno dimljene pastrmke",
volume = "61",
number = "4",
pages = "429-442",
doi = "10.2298/AVB1104429D"
}

Dimitrijević, M., Anderson, R. C., Karabasil, N., Pavlićević, N., Jovanović, S., Nedeljković-Trailović, J., Teodorović, V., Marković, M.,& Dojčinović, S.. (2011). Environmental prevalence and persistence of Listeria monocytogenes in cold-smoked trout processing plants. in Acta Veterinaria-Beograd
Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 61(4), 429-442.
https://doi.org/10.2298/AVB1104429D

Dimitrijević M, Anderson RC, Karabasil N, Pavlićević N, Jovanović S, Nedeljković-Trailović J, Teodorović V, Marković M, Dojčinović S. Environmental prevalence and persistence of Listeria monocytogenes in cold-smoked trout processing plants. in Acta Veterinaria-Beograd. 2011;61(4):429-442.
doi:10.2298/AVB1104429D .

Dimitrijević, Mirjana, Anderson, Robin C., Karabasil, Nedjeljko, Pavlićević, Nataša, Jovanović, S., Nedeljković-Trailović, Jelena, Teodorović, Vlado, Marković, Maja, Dojčinović, Slobodan, "Environmental prevalence and persistence of Listeria monocytogenes in cold-smoked trout processing plants" in Acta Veterinaria-Beograd, 61, no. 4 (2011):429-442,
https://doi.org/10.2298/AVB1104429D . .

TY  - JOUR
AU  - Stančić, Ivan
AU  - Stančić, Blagoje
AU  - Božić, Aleksandar
AU  - Anderson, Robin C.
AU  - Harvey, Roger B.
AU  - Gvozdić, Dragan
PY  - 2011
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/755
AB  - About 30% of the total number of gilts selected for reproduction at the large breeding farm units in Vojvodina (Republic of Serbia) are culled due to prolonged pre-insemination anoestrus (estrus not detected until 8 mo of age). The aim of this study was to provide the answer to the following question: do the culling gilts reach cyclic ovarian activity at all? One hundred seventy five culled gilts in which external estrus manifestations were not detected by 8 mo of age were sacrificed and their reproductive organs were examined for determination of sexual maturity (ovaries exhibiting pre-ovulatory follicles 8 to 11 mm in diameter, corpora hemorrhagica, corpora lutea and corpora albicantia). Uterine weights and horn length were also determined. Functional ovaries were observed in 107 (61.1%) examined gilts, with 62 animals having one and 45 having two puberty ovarian cycles (57.9% and 42.1%, respectively). Pathomorphological changes which could result in prolonged pre-insemination anoestrus were not observed on the reproductive organs of sexually mature gilts. Our results indicate that most of the culling gilts have reached cyclic ovarian activity. The main reason for culling due to the absence of external estrus manifestations in sexually mature gilts could be inadequate estrus detection technology.
PB  - Elsevier Science Inc, New York
T2  - Theriogenology
T1  - Ovarian activity and uterus organometry in delayed puberty gilts
VL  - 76
IS  - 6
SP  - 1022
EP  - 1026
DO  - 10.1016/j.theriogenology.2011.05.004
ER  - 

@article{
author = "Stančić, Ivan and Stančić, Blagoje and Božić, Aleksandar and Anderson, Robin C. and Harvey, Roger B. and Gvozdić, Dragan",
year = "2011",
abstract = "About 30% of the total number of gilts selected for reproduction at the large breeding farm units in Vojvodina (Republic of Serbia) are culled due to prolonged pre-insemination anoestrus (estrus not detected until 8 mo of age). The aim of this study was to provide the answer to the following question: do the culling gilts reach cyclic ovarian activity at all? One hundred seventy five culled gilts in which external estrus manifestations were not detected by 8 mo of age were sacrificed and their reproductive organs were examined for determination of sexual maturity (ovaries exhibiting pre-ovulatory follicles 8 to 11 mm in diameter, corpora hemorrhagica, corpora lutea and corpora albicantia). Uterine weights and horn length were also determined. Functional ovaries were observed in 107 (61.1%) examined gilts, with 62 animals having one and 45 having two puberty ovarian cycles (57.9% and 42.1%, respectively). Pathomorphological changes which could result in prolonged pre-insemination anoestrus were not observed on the reproductive organs of sexually mature gilts. Our results indicate that most of the culling gilts have reached cyclic ovarian activity. The main reason for culling due to the absence of external estrus manifestations in sexually mature gilts could be inadequate estrus detection technology.",
publisher = "Elsevier Science Inc, New York",
journal = "Theriogenology",
title = "Ovarian activity and uterus organometry in delayed puberty gilts",
volume = "76",
number = "6",
pages = "1022-1026",
doi = "10.1016/j.theriogenology.2011.05.004"
}

Stančić, I., Stančić, B., Božić, A., Anderson, R. C., Harvey, R. B.,& Gvozdić, D.. (2011). Ovarian activity and uterus organometry in delayed puberty gilts. in Theriogenology
Elsevier Science Inc, New York., 76(6), 1022-1026.
https://doi.org/10.1016/j.theriogenology.2011.05.004

Stančić I, Stančić B, Božić A, Anderson RC, Harvey RB, Gvozdić D. Ovarian activity and uterus organometry in delayed puberty gilts. in Theriogenology. 2011;76(6):1022-1026.
doi:10.1016/j.theriogenology.2011.05.004 .

Stančić, Ivan, Stančić, Blagoje, Božić, Aleksandar, Anderson, Robin C., Harvey, Roger B., Gvozdić, Dragan, "Ovarian activity and uterus organometry in delayed puberty gilts" in Theriogenology, 76, no. 6 (2011):1022-1026,
https://doi.org/10.1016/j.theriogenology.2011.05.004 . .

TY  - JOUR
AU  - Kim, W.K.
AU  - Karabasil, Nedjeljko
AU  - Bulajić, Snežana
AU  - Dunkley, K.D.
AU  - Callaway, T.R.
AU  - Poole, T.L.
AU  - Ricke, S.C.
AU  - Anderson, Robin C.
AU  - Nisbet, David J.
PY  - 2005
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/342
AB  - The objective of the study was to determine the frequency of spontaneous acquisition of resistance to select antibiotics by Salmonella Typhimurium (ST) when grown in glucose amended continuous flow culture at slow (D = 0.025 h -1) or fast (D = 0.27 h-1) dilution rates. The bacterium was grown in LB minimal medium (pH 6.25) containing no antibiotics. Upon achieving steady state, samples were plated to tryptic soy agar (TSA) alone or supplemented (per ml) with 2 and 16 μg oxytetracycline, 4 and 16 μg tetracycline, 2 and 64 μg kanamycin, and 0.25 and 2 μg enrofloxacin. Regardless of growth rate, CFU of resistant ST from the TSA containing antibiotics was less than 2 × 101 except for 2 μg kanamycin and 0.25 μg enrofloxacin treatments (higher than 1 × 109 and 4 × 107 CFU of resistant ST for trials 1 and 2, respectively). Frequency of recovering resistant ST from the TSA containing the higher antibiotic concentrations was less than 1 in 109 for all antibiotics, but was higher on the media containing 2 μg kanamycin and 0.25 μg enrofloxacin at both slow and fast growth rates. In general, minimal susceptibility differences were detected for isolates from slow and fast dilution rates.
PB  - Taylor and Francis Inc.
T2  - Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural
T1  - Comparison of spontaneous antibiotic resistance frequency of Salmonella typhimurium growth in glucose amended continuous culture at slow and fast dilution rates
VL  - 40
IS  - 3
SP  - 475
EP  - 484
DO  - 10.1081/PFC-200047598
ER  - 

@article{
author = "Kim, W.K. and Karabasil, Nedjeljko and Bulajić, Snežana and Dunkley, K.D. and Callaway, T.R. and Poole, T.L. and Ricke, S.C. and Anderson, Robin C. and Nisbet, David J.",
year = "2005",
abstract = "The objective of the study was to determine the frequency of spontaneous acquisition of resistance to select antibiotics by Salmonella Typhimurium (ST) when grown in glucose amended continuous flow culture at slow (D = 0.025 h -1) or fast (D = 0.27 h-1) dilution rates. The bacterium was grown in LB minimal medium (pH 6.25) containing no antibiotics. Upon achieving steady state, samples were plated to tryptic soy agar (TSA) alone or supplemented (per ml) with 2 and 16 μg oxytetracycline, 4 and 16 μg tetracycline, 2 and 64 μg kanamycin, and 0.25 and 2 μg enrofloxacin. Regardless of growth rate, CFU of resistant ST from the TSA containing antibiotics was less than 2 × 101 except for 2 μg kanamycin and 0.25 μg enrofloxacin treatments (higher than 1 × 109 and 4 × 107 CFU of resistant ST for trials 1 and 2, respectively). Frequency of recovering resistant ST from the TSA containing the higher antibiotic concentrations was less than 1 in 109 for all antibiotics, but was higher on the media containing 2 μg kanamycin and 0.25 μg enrofloxacin at both slow and fast growth rates. In general, minimal susceptibility differences were detected for isolates from slow and fast dilution rates.",
publisher = "Taylor and Francis Inc.",
journal = "Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural",
title = "Comparison of spontaneous antibiotic resistance frequency of Salmonella typhimurium growth in glucose amended continuous culture at slow and fast dilution rates",
volume = "40",
number = "3",
pages = "475-484",
doi = "10.1081/PFC-200047598"
}

Kim, W.K., Karabasil, N., Bulajić, S., Dunkley, K.D., Callaway, T.R., Poole, T.L., Ricke, S.C., Anderson, R. C.,& Nisbet, D. J.. (2005). Comparison of spontaneous antibiotic resistance frequency of Salmonella typhimurium growth in glucose amended continuous culture at slow and fast dilution rates. in Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural
Taylor and Francis Inc.., 40(3), 475-484.
https://doi.org/10.1081/PFC-200047598

Kim W, Karabasil N, Bulajić S, Dunkley K, Callaway T, Poole T, Ricke S, Anderson RC, Nisbet DJ. Comparison of spontaneous antibiotic resistance frequency of Salmonella typhimurium growth in glucose amended continuous culture at slow and fast dilution rates. in Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural. 2005;40(3):475-484.
doi:10.1081/PFC-200047598 .

Kim, W.K., Karabasil, Nedjeljko, Bulajić, Snežana, Dunkley, K.D., Callaway, T.R., Poole, T.L., Ricke, S.C., Anderson, Robin C., Nisbet, David J., "Comparison of spontaneous antibiotic resistance frequency of Salmonella typhimurium growth in glucose amended continuous culture at slow and fast dilution rates" in Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural, 40, no. 3 (2005):475-484,
https://doi.org/10.1081/PFC-200047598 . .