Araştırma Makalesi
Presence and virulence characterization of Listeria monocytogenes from fish samples in the Black Sea, Turkey
Öz
Listeria monocytogenes, characterized by a high mortality rate in humans, is a bacterium that causes listeriosis and is found in various aquatic products. The aim of this research was to investigate the presence, serotype distribution, virulence factor genes, and antibiotic susceptibility of L. monocytogenes strains isolated from a total of 500 fish samples of whiting (Merlangius merlangus euxinus) (n:243) and striped red mullet (Mullus surmuletus) (n:257) caught in the Black Sea between the years 2013-2014. Only one (0.2%) L. monocytogenes strain (striped red mullet) was isolated according to the cultural method (EN ISO 11290-1) and confirmed by PCR analysis. The L. monocytogenes strain was identified as serogroup 4b-4d-4e. Furthermore, the strain harboured hlyA, inlA, inlC, inlJ, plcA, plcB, prfA, mpl, actA, monoA-B, flaA, lip 1-2a, fri, iap, and gtcA genes except the dltA gene. On the other hand, L. monocytogenes strain susceptibility to ampicillin, meropenem, erythromycin, trimetophrim/sulfamethoxazole, and penicillin G was evaluated with the disc-diffusion method. According to the results, serogroup 4b-4d-4e isolated from striped red mullet was found to be unique to raw fish and susceptible to all tested antibiotics. In addition, it is considered that carrying out this research in different seas and with different fish species would be appropriate for determining the prevalence and virulence characteristics of L. monocytogenes.
Anahtar Kelimeler
Antibiotic susceptibility fish Listeria monocytogenes virulence genes
Destekleyen Kurum
This study was supported by the Scientific Research Project Fund of Istanbul University-Cerrahpaşa with the Project number of 56827.
Proje Numarası
56827
Teşekkür
This study is produced from the first author’s PhD thesis.
Kaynakça
- Akkaya L, Atabay HI, Gok V, et al (2011): Detection of Listeria species in fresh fish and fish market environment by IMS technique in Turkey. J Food Saf Food Qual, 62, 1-32.
- Bonazzi M, Lecuit M, Cossart P (2009): Listeria monocytogenes internalin and E-cadherin: From bench to bedside. Csh Perspect Biol, 1, a003087.
- Bubert A, Marcusrauch I, Lehner A, et al (1999): Detection and Differentiation of Listeria spp. by a Single Reaction Based on Multiplex PCR. Appl Environ Microbiol, 65, 4688-4692.
- Cabello FC (2006): Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment. Environ Microbiol, 8, 1137-1144.
- D’agostino M, Wagner M, Vazquez-Boland JA, et al (2004): A Validated PCR-Based Method to Detect Listeria monocytogenes Using Raw Milk as a Food Model-Towards an International Standart. J Food Protect, 67, 1646-1655.
- Dhanashree B, Otta SK, Karunasagar I, et al (2003): Incidence of Listeria spp. in clinical and food samples in Mangalore, India. Food Microbiol, 20, 447-453.
- Doumith M, Buchrieser C, Glaser P, et al (2004): Differentiation of the major Listeria monocytogenes serovars by multiplex PCR. J Clin Microbiol, 42, 3819-3822.
- Dussurget O, Cabanes D, Dehoux P, et al (2002): Listeria monocytogenes bile salt hydrolase is a PrfA-regulated virulence factor involved in the intestinal and hepatic phases of listeriosis. Mol Microbiol, 45, 1095–1106.
- Dussurget O, Pizzaro-Cerda J, Cossart P (2004): Molecular determinants of Listeria monocytogenes virulence. Annu Rev Microbiol, 58, 587–610.
- EFSA/ECDC (2019): The European Union One Health 2018 Zoonoses Report. Available at https://www.efsa.europa.eu/en/efsajournal/pub/5926 (Accessed May 18, 2021).
- El Marrakchi A, Boum’handi N, Hamama A (2005): Performance of a new chromogenic plating medium for the isolation of Listeria monocytogenes from marine environments. Lett Appl Microbiol, 40, 87-91.
- Ertas HB, Seker E (2005): Isolation of Listeria monocytogenes from Fish Intestines and RAPD Analysis. Turk J Vet Anim Sci, 29, 1007-1011.
- EUCAST (2017): EUCAST disk diffusion method for antimicrobial susceptibility testing Version 6.0. Available at http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_test_documents/Version_5/Manual_v_6.0_EUCAST_Disk_Test_final.pdf (Accessed Feb 18, 2018).
- EUCAST (2017): Media preparation for EUCAST disk diffusion testing and for determination of MIC values by the broth microdilution method. Available at http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_test_documents/Version_5/Media_preparation_v_5.0_EUCAST_AST.pdf (Accessed Feb 18, 2018).
- EUCAST (2018): Breakpoint tables for interpretation of MICs and zone diameters Version 8.0, valid from 2018-01-01. Available at http://www.eucast.org/fileadmin/src/ media/PDFs/EUCAST_files/Breakpoint_tables/v_8.0_Breakpoint_Tables.pdf. (Accessed Feb 21, 2018).
- Fallah AA, Saei-Dehkordia SS, Mahzouniehb M (2013): Occurrence and antibiotic resistance profiles of Listeria monocytogenes isolated from seafood products and market and processing environments in Iran. Food Control, 34, 630-636.
- GDFA (2013): Statistical Area of Aquatic Products.
- GDFA (2020): Su Ürünleri İstatistikleri Available at https://www.tarimorman.gov.tr/BSGM/Belgeler/Icerikler/Su%20%C3%9Cr%C3%BCnleri%20Veri%20ve%20D%C3%B6k%C3%BCmanlar%C4%B1/Su-%C3%9Cr%C3 %BCnleri-%C4%B0statistikleri.pdf (Accessed May 18, 2021).
- ISO (2004): Microbiology of food animal feeding stuffs: Horizontal method for the detection and enumeration of Listeria monocytogenes. Part 1: Detection method Amendment 1: Modification of isolation media and haemolysis test, and inclusion of precision data. ISO 11290-1/A1, Geneva, Switzerland.
- Jamali H, Paydar M, Ismail S, et al (2015): Prevalence, antimicrobial susceptibility and virulotyping of Listeria species and Listeria monocytogenes isolated from open-air fish markets. BMC Microbiol, 15, 144.
- Jaradat ZW, Schutze GE, Bhunia AK (2002): Genetic homogenity among Listeria monocytogenes strains from infected patients and meat products from two geographic locations determined by phenotyping, ribotyping and PCR analysis of virulence genes. Int J Food Microbiol, 76, 1-10.
- Kramarenko T, Roasto M, Meremӓe K, et al (2013): Listeria monocytogenes prevalence and serotype diversity in various foods. Food Control, 30, 24-29.
- Kyoui D, Takahashi H, Miya S, et al (2014): Comparison of the major virulence-related genes of Listeria monocytogenes in Internalin A truncated strain 36-25-1 and a clinical wild-type strain. BMC Microbiol, 14, 15.
- Leimeister-Wachter M, Domann E, Chakraborty T (1991): Detection of a gene encoding a phosphatidylinositol-specific phospholipase C that is co-ordinately expressed with listeriolysin in Listeria monocytogenes. Mol Microbiol, 5, 361-366.
- Liu D, Ainsworth AJ, Austin FW, et al (2004): Use of PCR primers derived from a putative transcriptional regulator gene for species-specific determination of Listeria monocytogenes. Int J Food Microbiol, 91, 297-304.
- Liu D, Lawrence ML, Austin FW et al (2007): A multiplex PCR for species- and virulence-specific determination of Listeria monocytogenes. J Microbiol Meth, 71, 133-140.
- Lomonaco S, Nucera D, Filipello V (2015): The evolution and epidemiology of Listeria monocytogenes in Europe and the United States. Infect Genet Evol, 35, 172-183.
- Lyautey E, Lapen DR, Wilkes G, et al (2007): Distribution and characteristics of Listeria monocytogenes isolates from surface waters of the South nation river watershed, Ontario, Canada. Appl Environ Microbiol, 73, 5401-5410.
- Marian MN, Sharifah Aminah SM, Zuraini MI, et al (2012): MPN-PCR detection and antimicrobial resistance of Listeria monocytogenes isolated from raw and ready-to-eat foods in Malaysia. Food Control, 28, 309-314.
- Mengaud J, Vincente MF, Cossart P (1989): Transcriptional mapping and nucleotide sequence of the Listeria monocytogenes hlyA region reveal structural features that may be involved in regulation. Infect Immun, 57, 3695-3701.
- Mizan FR, Jahid IK, Ha, S-H (2015): Microbial biofilms in seafood: A food-hygiene challenge. Food Microbiol, 49, 41-55.
- Molla B, Yilma R, Alemayehu D (2004): Listeria monocytogenes and other Listeria species in retail meat and milk products in Addis Adaba, Ethiopia. Ethiop J Health Dev, 18, 208-212.
- Nishibori T, Cooray K, Xiong H, et al (1995): Correlation between the Presence of Virulence-Associated Genes as Determined by PCR and Actual Virulence to Mice in Various Strains of Listeria Spp. Microbiol Immunol, 39, 343-349.
- Orsi RH, Wiedmann M (2016): Characteristics and distribution of Listeria spp., including Listeria species newly described since 2009. Appl Microbiol Biotechnol, 12, 5273-5287.
- Promadej N, Fielder F, Cossart P, et al (1999): Cell Wall Teichoic Acid Glycosylation in Listeria monocytogenes Serotype 4b Requires gtcA, a novel, Serogrup-Specific Gene. J Bacteriol, 181, 418-425.
- Rodάs-Suάrez OR, Flores-Pedroche JF, Betancourt-Rule JM, et al (2006): Occurrence and antibiotic sensitivity of Listeria monocytogenes strains isolated from oysters, fish, and estuarine water. Appl Environ Microbiol, 72, 7410-7412.
- Shen J, Rump L, Zhang Y, et al (2013): Molecular subtyping and virulence gene analysis of Listeria monocytogenes isolates from food. Food Microbiol, 35, 58-64.
- Siriken B, Ayaz ND, Erol I (2013): Prevalence and serotype distribution of Listeria monocytogenes in salted anchovy, raw anchovy, and raw mussel using IMS-based cultivation technique and PCR. J Aquat Food Prod T, 22, 77-82.
- Skowron K, Kwiecińska-Pirόg J, Grudlewska K, et al (2018): The occurence, transmission, virulence and antibiotic resistance of Listeria monocytogenes in fish processin plant. Int J Food Microbiol, 282, 71-83.
- Slama RB, Miladi H, Chaieb K, et al (2013): Survival of Listeria monocytogenes Cells and the Effects of Extended Frozen Storage (-20°C) on the Expression of Its Virulence Gene. Appl Biochem Biotech, 170, 1174-1183.
- Soultos N, Abrahim A, Papageorgiou K, et al (2007): Incidence of Listeria spp. in fish and environment of fish markets in northern Greece. Food Control, 18, 554-557.
- Stea EC, Purdue LM, Jamieson RC, et al (2015): Comparison of the Prevalences and Diversities of Listeria Species and Listeria monocytogenes in an Urban and a Rural Agricultural Watershed. Appl Environ Microbiol, 81, 3812-3822.
- TAGEM (2019): Su ürünleri Sektör Politika Belgesi 2019-2023. Available at https: //www.tarimorman.gov.tr/ TAGEM/Belgeler/yayin/Su%20Ürünleri%20Sektör%20Politika%20Belgesi%202019-2023.pdf (Accessed May 18, 2021).
- Thomas DJI, Strachan N, Goodburn K, et al (2012): A review of the published literatüre and current production and processing practices in smoked fish processing plants with emphasis on contamination by Listeria monocytogenes. Aberdeen (UK): Food Standards Agency. Report No: Final FSA report. Available at http://www.foodstandards.gov.scot/downloads/Final_Report_%285%29.pdf (Accessed Jun 06, 2018).
- Ucun Ozel H, Gemici B (2016): Bartın Irmağı Kirlilik Profilinin Fiziksel Parametrelerle Belirlenmesi. Mak Er U Fen Bil Ens Derg, 7, 52-58.
- Vasilev V, Japheth R, Breuer R, et al (2010): A survey of Listeria monocytogenes strains, isolated from ready-to-eat foods in Israel over a period of 10 years, 1998-2007. Food Control, 21, 1179-1181.
- Vázquez-Boland JA, Kocks C, Dramsi S, et al (1992): Nucleotide Sequence of the Lecithinase Operon of Listeria monocytogenes and Possible Role of Lecitinase in Cell-to-Cell Spread. Infect Immun, 60, 219-230.
- Wang X, Lü X, Yin L, et al (2013): Occurrence and antimicrobial susceptibility of Listeria monocytogenes isolates from retail raw foods. Food Control, 32, 153-158.
- Wieczorek K, Osek J (2017): Prevalence, genetic diversity and antimicrobial resistance of Listeria monocytogenes isolated from fresh and smoked fish in Poland. Food Microbiol, 64, 164-171.
- Yucel N, Balci S (2010): Prevalence of Listeria, Aeromonas, and Vibrio species in fish used for human consumption in Turkey. J Food Protect, 73, 380-384.
Yıl 2022,
Cilt: 69 Sayı: 4, 387 - 394, 30.09.2022
Öz
Proje Numarası
56827
Kaynakça
- Akkaya L, Atabay HI, Gok V, et al (2011): Detection of Listeria species in fresh fish and fish market environment by IMS technique in Turkey. J Food Saf Food Qual, 62, 1-32.
- Bonazzi M, Lecuit M, Cossart P (2009): Listeria monocytogenes internalin and E-cadherin: From bench to bedside. Csh Perspect Biol, 1, a003087.
- Bubert A, Marcusrauch I, Lehner A, et al (1999): Detection and Differentiation of Listeria spp. by a Single Reaction Based on Multiplex PCR. Appl Environ Microbiol, 65, 4688-4692.
- Cabello FC (2006): Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment. Environ Microbiol, 8, 1137-1144.
- D’agostino M, Wagner M, Vazquez-Boland JA, et al (2004): A Validated PCR-Based Method to Detect Listeria monocytogenes Using Raw Milk as a Food Model-Towards an International Standart. J Food Protect, 67, 1646-1655.
- Dhanashree B, Otta SK, Karunasagar I, et al (2003): Incidence of Listeria spp. in clinical and food samples in Mangalore, India. Food Microbiol, 20, 447-453.
- Doumith M, Buchrieser C, Glaser P, et al (2004): Differentiation of the major Listeria monocytogenes serovars by multiplex PCR. J Clin Microbiol, 42, 3819-3822.
- Dussurget O, Cabanes D, Dehoux P, et al (2002): Listeria monocytogenes bile salt hydrolase is a PrfA-regulated virulence factor involved in the intestinal and hepatic phases of listeriosis. Mol Microbiol, 45, 1095–1106.
- Dussurget O, Pizzaro-Cerda J, Cossart P (2004): Molecular determinants of Listeria monocytogenes virulence. Annu Rev Microbiol, 58, 587–610.
- EFSA/ECDC (2019): The European Union One Health 2018 Zoonoses Report. Available at https://www.efsa.europa.eu/en/efsajournal/pub/5926 (Accessed May 18, 2021).
- El Marrakchi A, Boum’handi N, Hamama A (2005): Performance of a new chromogenic plating medium for the isolation of Listeria monocytogenes from marine environments. Lett Appl Microbiol, 40, 87-91.
- Ertas HB, Seker E (2005): Isolation of Listeria monocytogenes from Fish Intestines and RAPD Analysis. Turk J Vet Anim Sci, 29, 1007-1011.
- EUCAST (2017): EUCAST disk diffusion method for antimicrobial susceptibility testing Version 6.0. Available at http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_test_documents/Version_5/Manual_v_6.0_EUCAST_Disk_Test_final.pdf (Accessed Feb 18, 2018).
- EUCAST (2017): Media preparation for EUCAST disk diffusion testing and for determination of MIC values by the broth microdilution method. Available at http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Disk_test_documents/Version_5/Media_preparation_v_5.0_EUCAST_AST.pdf (Accessed Feb 18, 2018).
- EUCAST (2018): Breakpoint tables for interpretation of MICs and zone diameters Version 8.0, valid from 2018-01-01. Available at http://www.eucast.org/fileadmin/src/ media/PDFs/EUCAST_files/Breakpoint_tables/v_8.0_Breakpoint_Tables.pdf. (Accessed Feb 21, 2018).
- Fallah AA, Saei-Dehkordia SS, Mahzouniehb M (2013): Occurrence and antibiotic resistance profiles of Listeria monocytogenes isolated from seafood products and market and processing environments in Iran. Food Control, 34, 630-636.
- GDFA (2013): Statistical Area of Aquatic Products.
- GDFA (2020): Su Ürünleri İstatistikleri Available at https://www.tarimorman.gov.tr/BSGM/Belgeler/Icerikler/Su%20%C3%9Cr%C3%BCnleri%20Veri%20ve%20D%C3%B6k%C3%BCmanlar%C4%B1/Su-%C3%9Cr%C3 %BCnleri-%C4%B0statistikleri.pdf (Accessed May 18, 2021).
- ISO (2004): Microbiology of food animal feeding stuffs: Horizontal method for the detection and enumeration of Listeria monocytogenes. Part 1: Detection method Amendment 1: Modification of isolation media and haemolysis test, and inclusion of precision data. ISO 11290-1/A1, Geneva, Switzerland.
- Jamali H, Paydar M, Ismail S, et al (2015): Prevalence, antimicrobial susceptibility and virulotyping of Listeria species and Listeria monocytogenes isolated from open-air fish markets. BMC Microbiol, 15, 144.
- Jaradat ZW, Schutze GE, Bhunia AK (2002): Genetic homogenity among Listeria monocytogenes strains from infected patients and meat products from two geographic locations determined by phenotyping, ribotyping and PCR analysis of virulence genes. Int J Food Microbiol, 76, 1-10.
- Kramarenko T, Roasto M, Meremӓe K, et al (2013): Listeria monocytogenes prevalence and serotype diversity in various foods. Food Control, 30, 24-29.
- Kyoui D, Takahashi H, Miya S, et al (2014): Comparison of the major virulence-related genes of Listeria monocytogenes in Internalin A truncated strain 36-25-1 and a clinical wild-type strain. BMC Microbiol, 14, 15.
- Leimeister-Wachter M, Domann E, Chakraborty T (1991): Detection of a gene encoding a phosphatidylinositol-specific phospholipase C that is co-ordinately expressed with listeriolysin in Listeria monocytogenes. Mol Microbiol, 5, 361-366.
- Liu D, Ainsworth AJ, Austin FW, et al (2004): Use of PCR primers derived from a putative transcriptional regulator gene for species-specific determination of Listeria monocytogenes. Int J Food Microbiol, 91, 297-304.
- Liu D, Lawrence ML, Austin FW et al (2007): A multiplex PCR for species- and virulence-specific determination of Listeria monocytogenes. J Microbiol Meth, 71, 133-140.
- Lomonaco S, Nucera D, Filipello V (2015): The evolution and epidemiology of Listeria monocytogenes in Europe and the United States. Infect Genet Evol, 35, 172-183.
- Lyautey E, Lapen DR, Wilkes G, et al (2007): Distribution and characteristics of Listeria monocytogenes isolates from surface waters of the South nation river watershed, Ontario, Canada. Appl Environ Microbiol, 73, 5401-5410.
- Marian MN, Sharifah Aminah SM, Zuraini MI, et al (2012): MPN-PCR detection and antimicrobial resistance of Listeria monocytogenes isolated from raw and ready-to-eat foods in Malaysia. Food Control, 28, 309-314.
- Mengaud J, Vincente MF, Cossart P (1989): Transcriptional mapping and nucleotide sequence of the Listeria monocytogenes hlyA region reveal structural features that may be involved in regulation. Infect Immun, 57, 3695-3701.
- Mizan FR, Jahid IK, Ha, S-H (2015): Microbial biofilms in seafood: A food-hygiene challenge. Food Microbiol, 49, 41-55.
- Molla B, Yilma R, Alemayehu D (2004): Listeria monocytogenes and other Listeria species in retail meat and milk products in Addis Adaba, Ethiopia. Ethiop J Health Dev, 18, 208-212.
- Nishibori T, Cooray K, Xiong H, et al (1995): Correlation between the Presence of Virulence-Associated Genes as Determined by PCR and Actual Virulence to Mice in Various Strains of Listeria Spp. Microbiol Immunol, 39, 343-349.
- Orsi RH, Wiedmann M (2016): Characteristics and distribution of Listeria spp., including Listeria species newly described since 2009. Appl Microbiol Biotechnol, 12, 5273-5287.
- Promadej N, Fielder F, Cossart P, et al (1999): Cell Wall Teichoic Acid Glycosylation in Listeria monocytogenes Serotype 4b Requires gtcA, a novel, Serogrup-Specific Gene. J Bacteriol, 181, 418-425.
- Rodάs-Suάrez OR, Flores-Pedroche JF, Betancourt-Rule JM, et al (2006): Occurrence and antibiotic sensitivity of Listeria monocytogenes strains isolated from oysters, fish, and estuarine water. Appl Environ Microbiol, 72, 7410-7412.
- Shen J, Rump L, Zhang Y, et al (2013): Molecular subtyping and virulence gene analysis of Listeria monocytogenes isolates from food. Food Microbiol, 35, 58-64.
- Siriken B, Ayaz ND, Erol I (2013): Prevalence and serotype distribution of Listeria monocytogenes in salted anchovy, raw anchovy, and raw mussel using IMS-based cultivation technique and PCR. J Aquat Food Prod T, 22, 77-82.
- Skowron K, Kwiecińska-Pirόg J, Grudlewska K, et al (2018): The occurence, transmission, virulence and antibiotic resistance of Listeria monocytogenes in fish processin plant. Int J Food Microbiol, 282, 71-83.
- Slama RB, Miladi H, Chaieb K, et al (2013): Survival of Listeria monocytogenes Cells and the Effects of Extended Frozen Storage (-20°C) on the Expression of Its Virulence Gene. Appl Biochem Biotech, 170, 1174-1183.
- Soultos N, Abrahim A, Papageorgiou K, et al (2007): Incidence of Listeria spp. in fish and environment of fish markets in northern Greece. Food Control, 18, 554-557.
- Stea EC, Purdue LM, Jamieson RC, et al (2015): Comparison of the Prevalences and Diversities of Listeria Species and Listeria monocytogenes in an Urban and a Rural Agricultural Watershed. Appl Environ Microbiol, 81, 3812-3822.
- TAGEM (2019): Su ürünleri Sektör Politika Belgesi 2019-2023. Available at https: //www.tarimorman.gov.tr/ TAGEM/Belgeler/yayin/Su%20Ürünleri%20Sektör%20Politika%20Belgesi%202019-2023.pdf (Accessed May 18, 2021).
- Thomas DJI, Strachan N, Goodburn K, et al (2012): A review of the published literatüre and current production and processing practices in smoked fish processing plants with emphasis on contamination by Listeria monocytogenes. Aberdeen (UK): Food Standards Agency. Report No: Final FSA report. Available at http://www.foodstandards.gov.scot/downloads/Final_Report_%285%29.pdf (Accessed Jun 06, 2018).
- Ucun Ozel H, Gemici B (2016): Bartın Irmağı Kirlilik Profilinin Fiziksel Parametrelerle Belirlenmesi. Mak Er U Fen Bil Ens Derg, 7, 52-58.
- Vasilev V, Japheth R, Breuer R, et al (2010): A survey of Listeria monocytogenes strains, isolated from ready-to-eat foods in Israel over a period of 10 years, 1998-2007. Food Control, 21, 1179-1181.
- Vázquez-Boland JA, Kocks C, Dramsi S, et al (1992): Nucleotide Sequence of the Lecithinase Operon of Listeria monocytogenes and Possible Role of Lecitinase in Cell-to-Cell Spread. Infect Immun, 60, 219-230.
- Wang X, Lü X, Yin L, et al (2013): Occurrence and antimicrobial susceptibility of Listeria monocytogenes isolates from retail raw foods. Food Control, 32, 153-158.
- Wieczorek K, Osek J (2017): Prevalence, genetic diversity and antimicrobial resistance of Listeria monocytogenes isolated from fresh and smoked fish in Poland. Food Microbiol, 64, 164-171.
- Yucel N, Balci S (2010): Prevalence of Listeria, Aeromonas, and Vibrio species in fish used for human consumption in Turkey. J Food Protect, 73, 380-384.
Toplam 50 adet kaynakça vardır.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Veteriner Cerrahi |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Proje Numarası | 56827 |
| Yayımlanma Tarihi | 30 Eylül 2022 |
| Yayımlandığı Sayı | Yıl 2022Cilt: 69 Sayı: 4 |
