Araştırma Makalesi
BibTex RIS Kaynak Göster

Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate

Yıl 2026, Sayı: Advanced Online Publication
https://doi.org/10.65206/pajes.44837

Öz

In this work, microbial inactivation experiments are performed on the biogas digestate samples via laboratory type ohmic heating system. The ohmic heating system ensures 2.72 V/cm voltage gradient with 50 Hz frequency square waveform pulsed alternating signals. Operations with 5 min and 15 min holding times are carried out at 70°C target temperature. Microbiological analysis performed before and after ohmic heating treatments with 5 minutes holding times revealed that, the number of Clostridium spp spores and vegetative cells suspended in the digestate with the related composition, are subjected to log reduction as 0.987 and 1.650 respectively. As a result of the 15 minutes holding time, it is determined that Clostridium spp spores and vegetative cells exhibited log reductions 0.819 and 1.725 respectively. The ohmic heating operation with 5 minutes holding time resulted in 18% decrease in the number of B.cereus spores and 55% increase in their vegetative cells in the same product. As a result of the 15 minutes holding time, the number of B.cereus spores and the vegetative cells increased by 37% and 23% respectively. Existing literature work on ohmic heating generally discusses the effects of the process parameters on the inactivation of microbial species which exist in various food matrixes. In this work, the effect of ohmic heating on the microbial inactivation of the biogas digestate which is originated from a biogas plant utilizing chicken manure as a substrate is investigated.

Kaynakça

  • [1] Tülek Y, Filizay G. “Applications of high hydrostatic pressure in the food industry”. Pamukkale University Journal of Engineering Sciences,12(1), 369–377, 2006.
  • [2] Keynan A, Evanchik Z, Halvorson HO, Hastings JW. “Activation of bacterial endospores”. J Bacteriol, 88(1), 313–8, 1964.
  • [3] Løvdal IS, Hovda MB, Granum PE, Rosnes JT. “Promoting Bacillus cereus spore germination for subsequent inactivation by mild heat treatment”. J Food Prot, 74(1), 2079–89,2011.
  • [4] Cho HY, Yousef AE, Sastry SK. “Kinetics of inactivation of Bacillus subtilis spores by continuous or intermittent ohmic and conventional heating”. Biotechnol Bioeng, 62(1), 368–72, 1999.
  • [5] Vepachedu VR, Hirneisen K, Hoover DG, Setlow P. “Studies of the release of small molecules during pressure germination of spores of Bacillus subtilis”. Lett Appl Microbiol, 45(1), 342–8, 2007.
  • [6] Somavat R, Mohamed HMH, Chung YK, Yousef AE, Sastry SK. “Accelerated inactivation of Geobacillus stearothermophilus spores by ohmic heating”. J Food Eng, 108(1), 69–76, 2012.
  • [7] Paredes-Sabja D, Setlow P, Sarker MR. “Germination of spores of Bacillales and Clostridiales species: mechanisms and proteins involved”. Trends Microbiol, 19(1), 85–94, 2011.
  • [8] Baysal AH, Icier F. “Inactivation Kinetics of Alicyclobacillus acidoterrestris Spores in orange juice by ohmic heating: effects of voltage gradient and temperature on inactivation”. J Food Prot, 73(1), 299–304, 2010.
  • [9] Blocher JC, Busta FF. “Bacterial spore resistance to acid”. Food Technology, 37(1), 87–89, 1983.
  • [10] Chang SS, Kang DH. “Alicyclobacillus spp. in the fruit juice industry: history, characteristics, and current isolation/detection procedures”. Crit Rev Microbiol, 30(1), 55–74, 2004.
  • [11] Zhang QH. “PEF treatment chamber designs and their effect on system configuration”. Institute of Food Technologists Annual Meeting, Atlanta, GA, 1998.
  • [12] Xiangxiao Qiu, Tuhela L, Zhang QH. “Application of pulsed power technology in nonthermal food processing and system optimization”, 11th IEEE International Pulsed Power Conference (Cat. No.97CH36127), USA, 1997.
  • [13] Demir E, Tappi S, Dymek K, Rocculi P, Gómez Galindo F. “Reversible electroporation caused by pulsed electric field – Opportunities and challenges for the food sector”. Trends in Food Science and Technology, 139(8), 104120, 2023.
  • [14] Novickij V, Rembiałkowska N, Szlasa W, Kulbacka J. “Does the shape of the electric pulse matter in electroporation?”. Front Oncol, 12(1), 958128, 2022.
  • [15] Wu WJ, Chang J. “Inactivation of vegetative cells, germinated spores, and dormant spores of Bacillus atrophaeus by pulsed electric field with fixed energy input”. J Food Process Eng, 45(2), 13959, 2022.
  • [16] Gould GW, Dring GJ. “Mechanisms of spore heat resistance”. Advances in Microbial Physiology, 11(1), 137-164, 1974.
  • [17] Grahl T, Märkl H. “Killing of microorganisms by pulsed electric fields”. Appl Microbiol Biotechnol, 45(1), 148–57, 1996.
  • [18] Jin, ZT, Su Y, Tuhela L, Zhang, QH, Sastry SK, Yousef AE. “Inactivation of Bacillus subtilis spores using high voltage pulsed electric fields”. Pulsed Electric Fields in Food Processing, 1, 167–81, 2019.
  • [19] Marquez VO, Mittal GS, Griffiths MW. “Destruction and inhibition of bacterial spores by high voltage pulsed electric field”. J Food Sci, 62(1), 399–401, 1997.
  • [20] Gerhardt P, Marquis RE, Spore thermoresistance measurements. Editors: Smith I, Slepecky RA, Setlow P. Regulation of Prokaryotic Development,43-63, Washington, USA, ASM Press, 1989.
  • [23] Yonemoto Y, Yamashita T, Muraji M, Tatebe W, Oshima H, Kato J.“Resistance of yeast and bacterial spores to high voltage electric pulses”. J Ferment Bioeng, 75(1), 99–102, 1993.
  • [24] Pillet F, Formosa-Dague C, Baaziz H, Dague E, Rols M-P. “Cell wall as a target for bacteria inactivation by pulsed electric fields”. Sci Rep, 6(1), 19778, 2016.
  • [25] Marsellés-Fontanet ÀR, Puig A, Olmos P, Mínguez-Sanz S, Martín-Belloso O. “Optimising the inactivation of grape juice spoilage organisms by pulse electric fields”. International Journal of Food Microbiology, 130(1), 159–65, 2009.
  • [26] Bermúdez-Aguirre D, Dunne CP, Barbosa-Cánovas GV. “Effect of processing parameters on inactivation of Bacillus cereus spores in milk using pulsed electric fields”. Int Dairy J, 24(1), 13–21, 2012.
  • [27] Eshtiaghi MN, Nakthong N. “Application of pulsed electric field for inactivation of Yeast S. cerevisiae in apple juice”. J Phys Conf Ser, 1893(1), 012008, 2021.
  • [28] Mendes-Oliveira G, Jin TZ, Campanella OH. “Modeling the inactivation of Escherichia coli O157:H7 and Salmonella Typhimurium in juices by pulsed electric fields: The role of the energy density”. J Food Eng, 282(1), 110001, 2020.
  • [29] Yan Z, Yin L, Hao C, Liu K, Qiu J. “Synergistic effect of pulsed electric fields and temperature on the inactivation of microorganisms”. AMB Express, 11(1), 47, 2021.
  • [30] Samaranayake CP, Sastry SK, Zhang H. “Pulsed Ohmic Heating– A novel technique for minimization of electrochemical reactions during processing”. J Food Sci, 70(8), 460-465, 2005.
  • [31] van Midden C, Harris J, Shaw L, et al., “The impact of anaerobic digestate on soil life: a review”. Applied Soil Ecology, 191(1), 2023.
  • [32] Froschle B, Heiermann M,Lebuhn M,Messelhausser U, Plochl M." Hygiene and Sanitation in Biogas Plants “. Adv Biochem Eng Biotechnol,151(1),63-99,2015.
  • [33] Ryang JH, Kim NH, Lee BS, Kim CT, Lee SH, Hwang IG. “Inactivation of Bacillus cereus spores in a tsuyu sauce using continuous ohmic heating with five sequential elbow-type electrodes”. J Appl Microbiol,120(1), 175–84, 2016.
  • [34] Sıkı R, Eryaşar A.”A literature review: Effects of ohmic heating on inactivation of specific bacterial spores in various matrixes”. Sigma Journal of Engineering and Natural Scienes, 43(4), 1100-1112, 2025.
  • [35] de Vries YP. “The role of calcium in bacterial spore germination”. Microbes Environ, 19(1), 3199–202, 2004.
  • [36] Harrell WK, Mantini E. “Studies on dipicolinic acid in the spores of Bacillus Cereus Var. Terminalis. Can”. J Microbiol, 3(5), 735–9,1957.
  • [37] Bishop HL, Doi RH. “Isolation and characterization of ribosomes from Bacillus subtilis spores”. J Bacteriol, 91(2), 695–701, 1966.

Kare vurgu dalgalı ohmik ısıtma işleminin, biyogaz prosesinden elde edilen fermente ürün içerisindeki B. Cereus ve Clostridium spp türlerinin canlı hücre ve sporları üzerindeki etkileri

Yıl 2026, Sayı: Advanced Online Publication
https://doi.org/10.65206/pajes.44837

Öz

Bu çalışmada, laboratuvar tipi ohmik ısıtma sistemi kullanılarak biyogaz santralinden alınan fermente ürün örneği üzerinde mikrobiyolojik inaktivasyon denemeleri gerçekleştirilmiştir. Ohmik ısıtma işleminde 50 Hz frekansında, 2.72 V/cm voltaj gradyanı sağlanan kare vurgu dalgalı alternatif sinyal kullanılmıştır. 70°C hedef sıcaklıkta, 5 dk ve 15 dk bekleme süreli işlemler gerçekleştirilmiştir. 5 dk bekleme süreli ohmik ısıtma işlemlerinin öncesi ve sonrasında gerçekleştirilen mikrobiyolojik analizler, ilgili bileşimdeki fermente ürün içerisindeki Clostridium spp spor ve canlı hücrelerinin sırasıyla 0.987 ve 1.650 log azalmaya tabi olduğunu göstermiştir. 15 dk bekleme süreli işlem neticesinde Clostridium spp spor ve canlı hücrelerinde sırasıyla 0.819 ve 1.725 log azalma tespit edilmiştir. 5 dk bekleme süreli ohmik ısıtma uygulaması neticesinde, aynı ürün içerisinde yer alan B.cereus spor sayısında %18 azalma, canlı hücre sayısında ise %55 artma tespit edilmiştir. 15 dk bekleme süreli işlem neticesinde B.cereus spor ve canlı hücre sayılarında sırasıyla %37 ve %23 artış meydana gelmiştir. Ohmik ısıtma kapsamındaki mevcut literatür çalışmaları genellikle, uygulanan proses parametrelerinin, çeşitli gıda ürünü matriksleri içerisinde bulunan mikrobiyal türlerin inaktivasyonu üzerine olan etkilerini tartışmaktadır. Bu çalışmada, ohmik ısıtma işleminin, proses girdisi tavuk gübresi olan bir biyogaz santralinden elde edilen fermente ürünün mikrobiyal inaktivasyonu üzerine olan etkisi araştırılmıştır.

Kaynakça

  • [1] Tülek Y, Filizay G. “Applications of high hydrostatic pressure in the food industry”. Pamukkale University Journal of Engineering Sciences,12(1), 369–377, 2006.
  • [2] Keynan A, Evanchik Z, Halvorson HO, Hastings JW. “Activation of bacterial endospores”. J Bacteriol, 88(1), 313–8, 1964.
  • [3] Løvdal IS, Hovda MB, Granum PE, Rosnes JT. “Promoting Bacillus cereus spore germination for subsequent inactivation by mild heat treatment”. J Food Prot, 74(1), 2079–89,2011.
  • [4] Cho HY, Yousef AE, Sastry SK. “Kinetics of inactivation of Bacillus subtilis spores by continuous or intermittent ohmic and conventional heating”. Biotechnol Bioeng, 62(1), 368–72, 1999.
  • [5] Vepachedu VR, Hirneisen K, Hoover DG, Setlow P. “Studies of the release of small molecules during pressure germination of spores of Bacillus subtilis”. Lett Appl Microbiol, 45(1), 342–8, 2007.
  • [6] Somavat R, Mohamed HMH, Chung YK, Yousef AE, Sastry SK. “Accelerated inactivation of Geobacillus stearothermophilus spores by ohmic heating”. J Food Eng, 108(1), 69–76, 2012.
  • [7] Paredes-Sabja D, Setlow P, Sarker MR. “Germination of spores of Bacillales and Clostridiales species: mechanisms and proteins involved”. Trends Microbiol, 19(1), 85–94, 2011.
  • [8] Baysal AH, Icier F. “Inactivation Kinetics of Alicyclobacillus acidoterrestris Spores in orange juice by ohmic heating: effects of voltage gradient and temperature on inactivation”. J Food Prot, 73(1), 299–304, 2010.
  • [9] Blocher JC, Busta FF. “Bacterial spore resistance to acid”. Food Technology, 37(1), 87–89, 1983.
  • [10] Chang SS, Kang DH. “Alicyclobacillus spp. in the fruit juice industry: history, characteristics, and current isolation/detection procedures”. Crit Rev Microbiol, 30(1), 55–74, 2004.
  • [11] Zhang QH. “PEF treatment chamber designs and their effect on system configuration”. Institute of Food Technologists Annual Meeting, Atlanta, GA, 1998.
  • [12] Xiangxiao Qiu, Tuhela L, Zhang QH. “Application of pulsed power technology in nonthermal food processing and system optimization”, 11th IEEE International Pulsed Power Conference (Cat. No.97CH36127), USA, 1997.
  • [13] Demir E, Tappi S, Dymek K, Rocculi P, Gómez Galindo F. “Reversible electroporation caused by pulsed electric field – Opportunities and challenges for the food sector”. Trends in Food Science and Technology, 139(8), 104120, 2023.
  • [14] Novickij V, Rembiałkowska N, Szlasa W, Kulbacka J. “Does the shape of the electric pulse matter in electroporation?”. Front Oncol, 12(1), 958128, 2022.
  • [15] Wu WJ, Chang J. “Inactivation of vegetative cells, germinated spores, and dormant spores of Bacillus atrophaeus by pulsed electric field with fixed energy input”. J Food Process Eng, 45(2), 13959, 2022.
  • [16] Gould GW, Dring GJ. “Mechanisms of spore heat resistance”. Advances in Microbial Physiology, 11(1), 137-164, 1974.
  • [17] Grahl T, Märkl H. “Killing of microorganisms by pulsed electric fields”. Appl Microbiol Biotechnol, 45(1), 148–57, 1996.
  • [18] Jin, ZT, Su Y, Tuhela L, Zhang, QH, Sastry SK, Yousef AE. “Inactivation of Bacillus subtilis spores using high voltage pulsed electric fields”. Pulsed Electric Fields in Food Processing, 1, 167–81, 2019.
  • [19] Marquez VO, Mittal GS, Griffiths MW. “Destruction and inhibition of bacterial spores by high voltage pulsed electric field”. J Food Sci, 62(1), 399–401, 1997.
  • [20] Gerhardt P, Marquis RE, Spore thermoresistance measurements. Editors: Smith I, Slepecky RA, Setlow P. Regulation of Prokaryotic Development,43-63, Washington, USA, ASM Press, 1989.
  • [23] Yonemoto Y, Yamashita T, Muraji M, Tatebe W, Oshima H, Kato J.“Resistance of yeast and bacterial spores to high voltage electric pulses”. J Ferment Bioeng, 75(1), 99–102, 1993.
  • [24] Pillet F, Formosa-Dague C, Baaziz H, Dague E, Rols M-P. “Cell wall as a target for bacteria inactivation by pulsed electric fields”. Sci Rep, 6(1), 19778, 2016.
  • [25] Marsellés-Fontanet ÀR, Puig A, Olmos P, Mínguez-Sanz S, Martín-Belloso O. “Optimising the inactivation of grape juice spoilage organisms by pulse electric fields”. International Journal of Food Microbiology, 130(1), 159–65, 2009.
  • [26] Bermúdez-Aguirre D, Dunne CP, Barbosa-Cánovas GV. “Effect of processing parameters on inactivation of Bacillus cereus spores in milk using pulsed electric fields”. Int Dairy J, 24(1), 13–21, 2012.
  • [27] Eshtiaghi MN, Nakthong N. “Application of pulsed electric field for inactivation of Yeast S. cerevisiae in apple juice”. J Phys Conf Ser, 1893(1), 012008, 2021.
  • [28] Mendes-Oliveira G, Jin TZ, Campanella OH. “Modeling the inactivation of Escherichia coli O157:H7 and Salmonella Typhimurium in juices by pulsed electric fields: The role of the energy density”. J Food Eng, 282(1), 110001, 2020.
  • [29] Yan Z, Yin L, Hao C, Liu K, Qiu J. “Synergistic effect of pulsed electric fields and temperature on the inactivation of microorganisms”. AMB Express, 11(1), 47, 2021.
  • [30] Samaranayake CP, Sastry SK, Zhang H. “Pulsed Ohmic Heating– A novel technique for minimization of electrochemical reactions during processing”. J Food Sci, 70(8), 460-465, 2005.
  • [31] van Midden C, Harris J, Shaw L, et al., “The impact of anaerobic digestate on soil life: a review”. Applied Soil Ecology, 191(1), 2023.
  • [32] Froschle B, Heiermann M,Lebuhn M,Messelhausser U, Plochl M." Hygiene and Sanitation in Biogas Plants “. Adv Biochem Eng Biotechnol,151(1),63-99,2015.
  • [33] Ryang JH, Kim NH, Lee BS, Kim CT, Lee SH, Hwang IG. “Inactivation of Bacillus cereus spores in a tsuyu sauce using continuous ohmic heating with five sequential elbow-type electrodes”. J Appl Microbiol,120(1), 175–84, 2016.
  • [34] Sıkı R, Eryaşar A.”A literature review: Effects of ohmic heating on inactivation of specific bacterial spores in various matrixes”. Sigma Journal of Engineering and Natural Scienes, 43(4), 1100-1112, 2025.
  • [35] de Vries YP. “The role of calcium in bacterial spore germination”. Microbes Environ, 19(1), 3199–202, 2004.
  • [36] Harrell WK, Mantini E. “Studies on dipicolinic acid in the spores of Bacillus Cereus Var. Terminalis. Can”. J Microbiol, 3(5), 735–9,1957.
  • [37] Bishop HL, Doi RH. “Isolation and characterization of ribosomes from Bacillus subtilis spores”. J Bacteriol, 91(2), 695–701, 1966.
Toplam 35 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Çevre Mühendisliği (Diğer)
Bölüm Araştırma Makalesi
Yazarlar

Rengin Sıkı

Ahmet Eryaşar

Gönderilme Tarihi 18 Nisan 2025
Kabul Tarihi 30 Ekim 2025
Erken Görünüm Tarihi 5 Aralık 2025
Yayımlandığı Sayı Yıl 2026 Sayı: Advanced Online Publication

Kaynak Göster

APA Sıkı, R., & Eryaşar, A. (2025). Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi(Advanced Online Publication). https://doi.org/10.65206/pajes.44837
AMA Sıkı R, Eryaşar A. Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi. Aralık 2025;(Advanced Online Publication). doi:10.65206/pajes.44837
Chicago Sıkı, Rengin, ve Ahmet Eryaşar. “Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate”. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi, sy. Advanced Online Publication (Aralık 2025). https://doi.org/10.65206/pajes.44837.
EndNote Sıkı R, Eryaşar A (01 Aralık 2025) Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi Advanced Online Publication
IEEE R. Sıkı ve A. Eryaşar, “Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate”, Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi, sy. Advanced Online Publication, Aralık2025, doi: 10.65206/pajes.44837.
ISNAD Sıkı, Rengin - Eryaşar, Ahmet. “Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate”. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi Advanced Online Publication (Aralık2025). https://doi.org/10.65206/pajes.44837.
JAMA Sıkı R, Eryaşar A. Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi. 2025. doi:10.65206/pajes.44837.
MLA Sıkı, Rengin ve Ahmet Eryaşar. “Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate”. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi, sy. Advanced Online Publication, 2025, doi:10.65206/pajes.44837.
Vancouver Sıkı R, Eryaşar A. Effects of square waveform pulsed ohmic heating on the vegetative cells and spores of B. cereus and Clostridium spp in the biogas digestate. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi. 2025(Advanced Online Publication).