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Presence of methicillin-resistant Staphylococcus aureus in slaughterhouse environment, pigs, carcasses, and workers Texte intégral
2017
Ivbule Meldra | Miklaševičs Edvīns | Čupāne Liene | Bērziņa Laima | Bālinš Andris | Valdovska Anda
Presence of methicillin-resistant Staphylococcus aureus in slaughterhouse environment, pigs, carcasses, and workers Texte intégral
2017
Ivbule Meldra | Miklaševičs Edvīns | Čupāne Liene | Bērziņa Laima | Bālinš Andris | Valdovska Anda
Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) is a highly resistant and difficult to cure zoonotic microorganism, which makes up a large part of food toxic infections and has shown high prevalence among pig population all over the world. The aim of the study was to establish the occurrence of MRSA in slaughterhouses, evaluate its antimicrobial resistance, and verify whether there are any differences or similarities with reference to other European countries. Material and Methods: A total of 100 pigs, 105 carcasses, 19 workers, and 24 samples from the environment of several slaughterhouses were examined by conventional microbial and molecular methods. Results: In total, 78 MRSA isolates were found. MRSA prevalence in slaughtered pigs varied from 8.0% to 88.6% depending on the slaughterhouse, reaching higher prevalence in slaughterhouses with higher slaughter capacity. In total, 21.1% of all workers were carriers of MRSA and 6.7% of carcasses were contaminated with MRSA. The 98.2% of MRSA isolates were resistant to penicillin, 89.1% to tetracycline, 60.1% to erythromycin, 65.5% to gentamycin, and 15 different spa types were found, among which spa type t01333 was most widespread. Conclusion: The study indicated that MRSA prevalence and spa types differed according to slaughterhouse slaughter capacity and good hygiene practices. Quite high MRSA occurrence among slaughterhouse workers is one of the main factors which increase pork contamination risk.
Afficher plus [+] Moins [-]Presence of methicillin-resistant Staphylococcus aureus in slaughterhouse environment, pigs, carcasses, and workers Texte intégral
2017
Ivbule, Meldra | Miklaševičs, Edvīns | Čupāne, Liene | Bērziņa, Laima | Bālinš, Andris | Valdovska, Anda
Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) is a highly resistant and difficult to cure zoonotic microorganism, which makes up a large part of food toxic infections and has shown high prevalence among pig population all over the world. The aim of the study was to establish the occurrence of MRSA in slaughterhouses, evaluate its antimicrobial resistance, and verify whether there are any differences or similarities with reference to other European countries. Material and Methods: A total of 100 pigs, 105 carcasses, 19 workers, and 24 samples from the environment of several slaughterhouses were examined by conventional microbial and molecular methods. Results: In total, 78 MRSA isolates were found. MRSA prevalence in slaughtered pigs varied from 8.0% to 88.6% depending on the slaughterhouse, reaching higher prevalence in slaughterhouses with higher slaughter capacity. In total, 21.1% of all workers were carriers of MRSA and 6.7% of carcasses were contaminated with MRSA. The 98.2% of MRSA isolates were resistant to penicillin, 89.1% to tetracycline, 60.1% to erythromycin, 65.5% to gentamycin, and 15 different spa types were found, among which spa type t01333 was most widespread. Conclusion: The study indicated that MRSA prevalence and spa types differed according to slaughterhouse slaughter capacity and good hygiene practices. Quite high MRSA occurrence among slaughterhouse workers is one of the main factors which increase pork contamination risk.
Afficher plus [+] Moins [-]Characterisation of newly emerged isolates of classical swine fever virus in China, 2014–2015 Texte intégral
2017
Leng Chaoliang | Zhang Hongliang | Kan Yunchao | Yao Lunguang | Li Mingliang | Zhai Hongyue | Li Zhen | Liu Chunxiao | Shi Hongfei | Ji Jun | Qiu Reng | Tian Zhijun
Characterisation of newly emerged isolates of classical swine fever virus in China, 2014–2015 Texte intégral
2017
Leng Chaoliang | Zhang Hongliang | Kan Yunchao | Yao Lunguang | Li Mingliang | Zhai Hongyue | Li Zhen | Liu Chunxiao | Shi Hongfei | Ji Jun | Qiu Reng | Tian Zhijun
Introduction: In 2014–2015, the epidemic of classical swine fever (CSF) occurred in many large-scale pig farms in different provinces of China, and a subgenotype 2.1d of CSF virus (CSFV) was newly identified.
Afficher plus [+] Moins [-]Characterisation of newly emerged isolates of classical swine fever virus in China, 2014–2015 Texte intégral
2017
Leng, Chaoliang | Zhang, Hongliang | Kan, Yunchao | Yao, Lunguang | Li, Mingliang | Zhai, Hongyue | Li, Zhen | Liu, Chunxiao | Shi, Hongfei | Ji, Jun | Qiu, Reng | Tian, Zhijun
Introduction: In 2014–2015, the epidemic of classical swine fever (CSF) occurred in many large-scale pig farms in different provinces of China, and a subgenotype 2.1d of CSF virus (CSFV) was newly identified. Material and Methods: The phylogenetic relationship, genetic diversity, and epidemic status of the 2014–2015 CSFV isolates, 18 new CSFV isolates collected in 2015, and 43 other strains isolated in 2014–2015 were fully analysed, together with 163 CSFV reference isolates. Results: Fifty-two 2014–2015 isolates belonged to subgenotype 2.1d and nine other isolates belonged to subgenotype 2.1b. The two subgenotype isolates showed unique molecular characteristics. Furthermore, the 2.1d isolates were found to possibly diverge from 2.1b isolates. Conclusion: This study suggests that the Chinese CSFVs will remain pandemic.
Afficher plus [+] Moins [-]Toxinotyping and antimicrobial resistance of Clostridium perfringens isolated from processed chicken meat products Texte intégral
2017
Hamza Dalia | Dorgham Sohad | Hakim Ashraf
Toxinotyping and antimicrobial resistance of Clostridium perfringens isolated from processed chicken meat products Texte intégral
2017
Hamza Dalia | Dorgham Sohad | Hakim Ashraf
Introduction: The toxinotyping and antimicrobial susceptibility of Clostridium perfringens strains isolated from processed chicken meat were determined.
Afficher plus [+] Moins [-]Toxinotyping and antimicrobial resistance of Clostridium perfringens isolated from processed chicken meat products Texte intégral
2017
Hamza, Dalia | Dorgham, Sohad | Ḥakīm, Ashraf
Introduction: The toxinotyping and antimicrobial susceptibility of Clostridium perfringens strains isolated from processed chicken meat were determined. Material and Methods: Two hundred processed chicken meat samples from luncheon meats, nuggets, burgers, and sausages were screened for Clostridium perfringens by multiplex PCR assay for the presence of alpha (cpa), beta (cpb), epsilon (etx), iota (ia), and enterotoxin toxin (cpe) genes. The C. perfringens isolates were examined in vitro against eight antibiotics (streptomycin, amoxicillin, ampicillin, ciprofloxacin, lincomycin, cefotaxime, rifampicin, and trimethoprim-sulfamethoxazole) Results: An overall of 32 C. perfringens strains (16%) were isolated from 200 processed chicken meat samples tested. The prevalence of C. perfringens was significantly dependent on the type of toxin genes detected (P = 0.0), being the highest in sausages (32%), followed by luncheon meats (24%), burgers (6%), and nuggets (2%). C. perfringens type A was the most frequently present toxinotype (24/32; 75%), followed by type D (21.9 %) and type E (3.1%). Of the 32 C. perfringens strains tested, only 9 (28%) were enterotoxin gene carriers, with most representing type A (n = 6). C. perfringens strains differed in their resistance/susceptibility to commonly used antibiotics. Most of the strains tested were sensitive to ampicillin (97%) and amoxicillin (94%), with 100% of the strains being resistant to streptomycin and lincomycin. It is noteworthy that the nine isolates with enterotoxigenic potential had a higher resistance than the non-enterotoxigenic ones. Conclusion: The considerably high C. perfringens isolation rates from processed chicken meat samples and resistance to some of the commonly used antibiotics indicate a potential public health risk. Recent information about the isolation of enterotoxigenic C. perfringens type E from chicken sausage has been reported.
Afficher plus [+] Moins [-]Development of a recombinant protein-based ELISA for detection of antibodies against bovine foamy virus Texte intégral
2017
Materniak-Kornas Magdalena | Osiński Zbigniew | Rudzki Marcin | Kuźmak Jacek
Development of a recombinant protein-based ELISA for detection of antibodies against bovine foamy virus Texte intégral
2017
Materniak-Kornas Magdalena | Osiński Zbigniew | Rudzki Marcin | Kuźmak Jacek
Introduction: Infections with bovine foamy virus (BFV) were found in many countries but there is a lack of large-scale surveys on the prevalence of BFV among dairy cattle. The aim of this study was to develop and validate the recombinant Gag protein-based ELISA and to estimate the prevalence of antibodies against BFV. Material and Methods: Gag coding region from BFV was cloned into expression vector pT7Arg-STOP, which expressed a high level of recombinant Gag protein from E.coli. The ELISA was standardised, and the cut-off value and sensitivity and specificity of the test were calculated using a receiver operating characteristic and Bayesian estimation. Results: A total of 3,051 serum samples were tested by ELISA and 939 (30.8%) sera were recognised as positive. When Bayesian approach was used, the overall true BFV prevalence was 29.7% (95% CI: 25.9-33.4%). Conclusion: Expressed Gag protein of BFV has been used successfully as an antigen for ELISA. Eventually, this study provides basic information about the epidemiological status of infection with BFV in dairy cattle in Poland, which can be used for further studies on dissemination and transmission of BFV infection.
Afficher plus [+] Moins [-]Development of a recombinant protein-based ELISA for detection of antibodies against bovine foamy virus Texte intégral
2017
Materniak-Kornas, Magdalena | Osiński, Zbigniew | Rudzki, Marcin | Kuźmak, Jacek
Introduction: Infections with bovine foamy virus (BFV) were found in many countries but there is a lack of large-scale surveys on the prevalence of BFV among dairy cattle. The aim of this study was to develop and validate the recombinant Gag protein-based ELISA and to estimate the prevalence of antibodies against BFV. Material and Methods: Gag coding region from BFV was cloned into expression vector pT7Arg-STOP, which expressed a high level of recombinant Gag protein from E.coli. The ELISA was standardised, and the cut-off value and sensitivity and specificity of the test were calculated using a receiver operating characteristic and Bayesian estimation. Results: A total of 3,051 serum samples were tested by ELISA and 939 (30.8%) sera were recognised as positive. When Bayesian approach was used, the overall true BFV prevalence was 29.7% (95% CI: 25.9-33.4%). Conclusion: Expressed Gag protein of BFV has been used successfully as an antigen for ELISA. Eventually, this study provides basic information about the epidemiological status of infection with BFV in dairy cattle in Poland, which can be used for further studies on dissemination and transmission of BFV infection.
Afficher plus [+] Moins [-]Epizootic and epidemiological situation of Trichinella sp. infection in Poland in 2006–2015 in view of wild boar population dynamics Texte intégral
2017
Flis Marian | Grela Eugeniusz R. | Gugała Dariusz
Epizootic and epidemiological situation of Trichinella sp. infection in Poland in 2006–2015 in view of wild boar population dynamics Texte intégral
2017
Flis Marian | Grela Eugeniusz R. | Gugała Dariusz
Introduction: The objective was to evaluate the epizootic and epidemiological situation of Trichinella sp. infection in Poland between 2006 and 2015 against the dynamics of the wild boar population and its primary reservoir host.
Afficher plus [+] Moins [-]Epizootic and epidemiological situation of Trichinella sp. infection in Poland in 2006–2015 in view of wild boar population dynamics Texte intégral
2017
Flis, Marian | Grela, Eugeniusz R. | Gugała, Dariusz
Introduction: The objective was to evaluate the epizootic and epidemiological situation of Trichinella sp. infection in Poland between 2006 and 2015 against the dynamics of the wild boar population and its primary reservoir host.Material and Methods: Boar and porcine trichinosis epizootic analysis was based on General Veterinary Inspectorate data from RRW-6 bulletins. The epidemiological situation was evaluated on the basis of the data supplied by the Department of Epidemiology of the National Institute of Hygiene - National Institute of Public Health. The wild boar hunting harvest and population dynamics were estimated, as these animals remain the basic infection source for humans. Population size and harvest data were obtained from hunting statistics.Results: The study timeframe showed an almost 2.5-fold increase in Trichinella infection cases in wild boars but a significant decline in human cases. In the domestic pig, the incidence rate did not exceed 0.00037%. The highest infection risk exists in West Pomerania, Greater Poland, and Kuyavian-Pomeranian Provinces. Over the study period, the wild boar population increased more than 1.5-fold, while the hunting harvest more than tripled. During the last two seasons the total hunt surpassed 100% of the spring population.Conclusion: Wild boar management by increasing the hunting take of the annual population growth should limit that growth and decrease the take in the future. Thereby, over some years intra-species trichinosis spread should reduce, for a substantial safety gain for wild boar meat.
Afficher plus [+] Moins [-]Evaluation of oxidative stress in sheep infected with Psoroptes ovis using total antioxidant capacity, total oxidant status, and malondialdehyde level Texte intégral
2017
Aktas Mustafa Sinan | Kandemir Fatih Mehmet | Kirbas Akin | Hanedan Basak | Aydin Mehmet Akif
Evaluation of oxidative stress in sheep infected with Psoroptes ovis using total antioxidant capacity, total oxidant status, and malondialdehyde level Texte intégral
2017
Aktas Mustafa Sinan | Kandemir Fatih Mehmet | Kirbas Akin | Hanedan Basak | Aydin Mehmet Akif
Introduction: The study aimed at evaluating oxidative stress using malondialdehyde (MDA), total antioxidant capacity (TAC), total oxidant status (TOS), and oxidative stress index (OSI) markers in sheep naturally infected with Psoroptes ovis (Acari).
Afficher plus [+] Moins [-]Evaluation of oxidative stress in sheep infected with Psoroptes ovis using total antioxidant capacity, total oxidant status, and malondialdehyde level Texte intégral
2017
Aktas, Mustafa Sinan | Kandemir, Fatih Mehmet | Kirbas, Akin | Hanedan, Basak | Aydın, M. Âkif
Introduction: The study aimed at evaluating oxidative stress using malondialdehyde (MDA), total antioxidant capacity (TAC), total oxidant status (TOS), and oxidative stress index (OSI) markers in sheep naturally infected with Psoroptes ovis (Acari).Material and Methods: The study was performed on 40 sheep divided into two equal groups: a healthy group (group I) and a group naturally infected with Psoroptes ovis (group II). The sera were obtained by centrifuging blood samples collected from the vena jugularis and serum MDA level changes in the samples were measured spectrophotometrically. Commercially available test kits were used for the measurement of TAC and TOS levels. The percentage ratio of TOS level to TAC level was accepted as OSI.Results: The serum malondialdehyde, total oxidant status levels, and oxidative stress index increased significantly (P < 0.01) in group II, while the serum total antioxidant capacity levels decreased significantly (P < 0.01) in this group. Negative correlations between total antioxidant capacity and total oxidant status and total antioxidant capacity and malondialdehyde, and a positive correlation between total oxidant status and malondialdehyde were found in infected sheep.Conclusion: The obtained results indicated the relationship between oxidant/antioxidant imbalance and Psoroptes ovis infection in sheep. Their MDA, TAC, TOS, and OSI markers may be used to determine the oxidative stress in natural infections with Psoroptes ovis.
Afficher plus [+] Moins [-]DON-induced changes in bone homeostasis in mink dams Texte intégral
2017
Tomaszewska Ewa | Dobrowolski Piotr | Muszyński Siemowit | Kostro Krzysztof | Taszkun Iwona | Żmuda Andrzej | Blicharski Tomasz | Hułas-Stasiak Monika
DON-induced changes in bone homeostasis in mink dams Texte intégral
2017
Tomaszewska Ewa | Dobrowolski Piotr | Muszyński Siemowit | Kostro Krzysztof | Taszkun Iwona | Żmuda Andrzej | Blicharski Tomasz | Hułas-Stasiak Monika
Introduction: The aim of the study was to investigate the mechanical and geometric properties as well as bone tissue and mineral density of long bones in mink dams exposed to deoxynivalenol (DON) since one day after mating, throughout gestation (ca. 46 d) and lactation to pelt harvesting. Material and Methods: Thirty clinically healthy multiparous minks (Neovison vison) of the standard dark brown type were used. After the mating, the minks were randomly assigned into two equal groups: nontreated control group and DON group fed wheat contaminated naturally with DON at a concentration of 1.1 mg·kg-1 of feed. Results: The final body weight and weight and length of the femur did not differ between the groups. However, DON contamination decreased mechanical endurance of the femur. Furthermore, DON reduced the mean relative wall thickness and vertical wall thickness of the femur, while vertical cortical index, midshaft volume, and cross-sectional moment of inertia increased. Finally, DON contamination did not alter bone tissue density, bone mineral density, or bone mineral content, but decreased the values of all investigated structural and material properties. Conclusion: DON at applied concentration probably intensified the process of endosteal resorption, which was the main reason for bone wall thinning and the weakening of the whole bone.
Afficher plus [+] Moins [-]DON-induced changes in bone homeostasis in mink dams Texte intégral
2017
Tomaszewska, Ewa | Dobrowolski, Piotr | Muszyński, Siemowit | Kostro, Krzysztof | Taszkun, Iwona | Żmuda, Andrzej | Blicharski, Tomasz | Hułas-Stasiak, Monika
Introduction: The aim of the study was to investigate the mechanical and geometric properties as well as bone tissue and mineral density of long bones in mink dams exposed to deoxynivalenol (DON) since one day after mating, throughout gestation (ca. 46 d) and lactation to pelt harvesting. Material and Methods: Thirty clinically healthy multiparous minks (Neovison vison) of the standard dark brown type were used. After the mating, the minks were randomly assigned into two equal groups: nontreated control group and DON group fed wheat contaminated naturally with DON at a concentration of 1.1 mg·kg⁻¹ of feed. Results: The final body weight and weight and length of the femur did not differ between the groups. However, DON contamination decreased mechanical endurance of the femur. Furthermore, DON reduced the mean relative wall thickness and vertical wall thickness of the femur, while vertical cortical index, midshaft volume, and cross-sectional moment of inertia increased. Finally, DON contamination did not alter bone tissue density, bone mineral density, or bone mineral content, but decreased the values of all investigated structural and material properties. Conclusion: DON at applied concentration probably intensified the process of endosteal resorption, which was the main reason for bone wall thinning and the weakening of the whole bone.
Afficher plus [+] Moins [-]Pathological alterations in respiratory system during co-infection with low pathogenic avian influenza virus (H9N2) and Escherichia coli in broiler chickens Texte intégral
2017
Jaleel Shahid | Younus Muhammad | Idrees Asif | Arshad Muhammad | Khan Aman Ullah | Ehtisham-ul-Haque Syed | Zaheer Muhammad Irfan | Tanweer Muhammad | Towakal Farhan | Munibullah | Tipu Muhammad Yasin | Sohail Muhammad Luqman | Umar Sajid
Pathological alterations in respiratory system during co-infection with low pathogenic avian influenza virus (H9N2) and Escherichia coli in broiler chickens Texte intégral
2017
Jaleel Shahid | Younus Muhammad | Idrees Asif | Arshad Muhammad | Khan Aman Ullah | Ehtisham-ul-Haque Syed | Zaheer Muhammad Irfan | Tanweer Muhammad | Towakal Farhan | Munibullah | Tipu Muhammad Yasin | Sohail Muhammad Luqman | Umar Sajid
Introduction: Despite the advancements in the field, there is a lack of data when it comes to co-infections in poultry. Therefore, this study was designed to address this issue. Material and Methods: Broiler birds were experimentally infected with E. coli (O78) and low pathogenic avian influenza (LPAI) strain, alone or in combination. The experimental groups were negative control. Results: The infected birds showed most severe clinical signs in E. coli+LPAI group along with a significant decrease in weight and enhanced macroscopic and microscopic pathological lesions. The survival rate was 60%, 84%, and 100% in birds inoculated with E. coli+LPAI, E. coli, and LPAI virus alone, respectively. The results showed that experimental co-infection with E. coli and H9N2 strain of LPAI virus increased the severity of clinical signs, mortality rate, and gross lesions. The HI titre against LPAI virus infection in the co-infected group was significantly higher than the HI titre of LPAI group, which may indicate that E. coli may promote propagation of H9N2 LPAI virus by alteration of immune response. Conclusion: The present study revealed that co-infection with E. coli and H9N2 LPAI virus caused more serious synergistic pathogenic effects and indicates the role of both pathogens as complicating factors in poultry infections.
Afficher plus [+] Moins [-]Pathological alterations in respiratory system during co-infection with low pathogenic avian influenza virus (H9N2) and Escherichia coli in broiler chickens Texte intégral
2017
Jaleel, Shahid | Younus, Muhammad | Idrees, Asif | Arshad, Muḥammad | Khan, Aman Ullah | Ehtisham-ul-Haque, Syed | Zaheer, Muhammad Irfan | Tanweer, Muhammad | Towakal, Farhan | Munibullah, | Tipu, Muhammad Yasin | Sohail, Muhammad Luqman | Umar, Sajid
Introduction: Despite the advancements in the field, there is a lack of data when it comes to co-infections in poultry. Therefore, this study was designed to address this issue. Material and Methods: Broiler birds were experimentally infected with E. coli (O78) and low pathogenic avian influenza (LPAI) strain, alone or in combination. The experimental groups were negative control. Results: The infected birds showed most severe clinical signs in E. coli+LPAI group along with a significant decrease in weight and enhanced macroscopic and microscopic pathological lesions. The survival rate was 60%, 84%, and 100% in birds inoculated with E. coli+LPAI, E. coli, and LPAI virus alone, respectively. The results showed that experimental co-infection with E. coli and H9N2 strain of LPAI virus increased the severity of clinical signs, mortality rate, and gross lesions. The HI titre against LPAI virus infection in the co-infected group was significantly higher than the HI titre of LPAI group, which may indicate that E. coli may promote propagation of H9N2 LPAI virus by alteration of immune response. Conclusion: The present study revealed that co-infection with E. coli and H9N2 LPAI virus caused more serious synergistic pathogenic effects and indicates the role of both pathogens as complicating factors in poultry infections.
Afficher plus [+] Moins [-]In ovo administration of CpG ODN induces expression of immune response genes in neonatal chicken spleen Texte intégral
2017
Sajewicz-Krukowska Joanna | Olszewska-Tomczyk Monika | Domańska-Blicharz Katarzyna
In ovo administration of CpG ODN induces expression of immune response genes in neonatal chicken spleen Texte intégral
2017
Sajewicz-Krukowska Joanna | Olszewska-Tomczyk Monika | Domańska-Blicharz Katarzyna
Introduction: Due to their immunostimulatory properties TLR ligands are used prophylactically to protect against a variety of viral and bacterial pathogens in mammals. Knowledge of the molecular and functional aspects of TLRs is essential for a better understanding of the immune system and resistance to diseases in birds. For that reason, this study attempted to determine the impact of TLR21 stimulation by its synthetic ligand (CpG ODN, class B) on the chicken immune system.
Afficher plus [+] Moins [-]In ovo administration of CpG ODN induces expression of immune response genes in neonatal chicken spleen Texte intégral
2017
Sajewicz-Krukowska, Joanna | Olszewska-Tomczyk, Monika | Domańska-Blicharz, Katarzyna
Introduction: Due to their immunostimulatory properties TLR ligands are used prophylactically to protect against a variety of viral and bacterial pathogens in mammals. Knowledge of the molecular and functional aspects of TLRs is essential for a better understanding of the immune system and resistance to diseases in birds. For that reason, this study attempted to determine the impact of TLR21 stimulation by its synthetic ligand (CpG ODN, class B) on the chicken immune system. Material and Methods: Sixty embryonated chicken eggs were randomly allocated into three groups (control and two experimental groups). On day 18 of embryonic development, chickens in one experimental group were administered in ovo a low dose of CpG ODN and the birds of the second experimental group were given a high dose of the ligand. Spleens were collected at 1, 2, 5, and 10 days post-hatching (dph) for analysis of IFN-α, IFN-β, IFN-γ, IL-6, and IL-10 expression using qRT-PCR. Results: Significant differences were observed in mRNA expression levels of all the measured cytokines associated with the modulation and regulation of the immune response at different time points. Conclusion: The obtained data clearly demonstrate that immune response induction takes place after in ovo administration of class B CpG ODN, and that the ligand has the ability to induce cytokine responses in neonatal chicken spleen.
Afficher plus [+] Moins [-]Development of LC-MS/MS confirmatory method for the determination of testosterone in bovine serum Texte intégral
2017
Woźniak Barbara | Matraszek-Żuchowska Iwona | Witek Sebastian | Posyniak Andrzej
Development of LC-MS/MS confirmatory method for the determination of testosterone in bovine serum Texte intégral
2017
Woźniak Barbara | Matraszek-Żuchowska Iwona | Witek Sebastian | Posyniak Andrzej
Introduction: In the European Union the use of steroid growth promoters is prohibited under Council Directive 96/22/EC. For effective control of illegal use of natural steroids, highly sensitive analytical methods are required, because sex hormones can be present in very low concentrations in biological samples. The aim of the study was to develop a confirmatory method for the detection of testosterone in bovine serum at ppt level.
Afficher plus [+] Moins [-]Development of LC-MS/MS confirmatory method for the determination of testosterone in bovine serum Texte intégral
2017
Woźniak-Sobczak, Barbara | Matraszek-Żuchowska, Iwona | Witek, Sebastian | Posyniak, Andrzej
Introduction: In the European Union the use of steroid growth promoters is prohibited under Council Directive 96/22/EC. For effective control of illegal use of natural steroids, highly sensitive analytical methods are required, because sex hormones can be present in very low concentrations in biological samples. The aim of the study was to develop a confirmatory method for the detection of testosterone in bovine serum at ppt level. Material and Methods: 17β-testosterone and internal standards of 17β-testosterone-d2 were extracted from serum samples with a mixture of tert-butyl methyl ether/petroleum ether and were directly analysed by an LC/MS/MS on QTRAP 5500 instrument with a TurboIon-Spray source operating in a positive ionisation mode. Chromatographic separation was achieved on the analytical column Inertsil® ODS-3 with an isocratic elution using mobile phase consisting of acetonitrile, methanol, and water. Method validation has been carried out in accordance with the Commission Decision 2002/657/EC. Results: The method was characterised by good recovery (82%) and precision (R.S.D 17 %). Decision limit (CCα) and detection capability (CCβ) was 0.05 μg L⁻¹ and 0.09 μg L⁻¹ respectively. The method met the criteria set out in Commission Decision 2002/657/EC for the purpose of confirmation in terms of retention time and ion ratio in the whole range of its application. Conclusions: The developed method is specific and sensitive, suitable for measuring the natural level of testosterone in blood of cattle and for use in routine control programme for the detection of this hormone in bovine serum.
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