“…However, this range is comparable to those of previous studies, which revealed that the range of most of the peaks is from 800-3500 Da (Edwards-Jones et al, 2000;Walker et al, 2002). As concluded by Shell et al (2017), on strains and isolates extracted from mastitis cases examined by MBT, the peaks ranged from 3,000 to 11,000 Da and the highest levels of intensity ranged between 4,000 and 10,000 Da. Such variations in the spectral peaks, range, and mass can be attributed to differences in the arrangement of sampling approaches.…”
The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infection is one of the greatest threats to both animal and human health. Our investigation was aimed to identify and differentiate between MRSA and methicillin-sensitive Staphylococcus aureus (MSSA) recovered from mastitic milk using MALDI-TOF mass spectrometry compared with phenotypic methods and studying their susceptibility to various antibiotics. Four hundred milk samples from mastitic animals (cows, sheep, goats, and dromedary camels) were investigated. Phenotypic identification of S. aureus was made through MASTASAPH Latex test, STAPH ID 32, and Vitek 2 system. The proteomic characterization of S. aureus was done by MBT. The Kirby Bauer method was accomplished to detect the resistance of S. aureus strains to antibiotics. The results of the MASTASAPH Latex test, revealed that 54 (46%) were recognized as S. aureus. All S. aureus isolates were identified by MBT with a score of more or equal 2.00. Several peaks were identified in the mass of 4590 Da, 4863 Da, and 4938 Da for MSSA and in the mass of 2636 Da and 3009 Da for MRSA. The MSP dendrogram demonstrated that the S. aureus isolates were classified into one group with a distance level of less or equal 400. The percentage of S. aureus resistance against carbenicillin, erythromycin and kanamycin was 94.4%, 38.88%, and 33.33%, respectively. In conclusion, S. aureus bacteria are among the key triggers for mastitis in Saudi Arabia. MBT is reported to be not only the rapid tool to identify S. aureus but also able to discriminate MRSA from MSSA.
“…However, this range is comparable to those of previous studies, which revealed that the range of most of the peaks is from 800-3500 Da (Edwards-Jones et al, 2000;Walker et al, 2002). As concluded by Shell et al (2017), on strains and isolates extracted from mastitis cases examined by MBT, the peaks ranged from 3,000 to 11,000 Da and the highest levels of intensity ranged between 4,000 and 10,000 Da. Such variations in the spectral peaks, range, and mass can be attributed to differences in the arrangement of sampling approaches.…”
The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infection is one of the greatest threats to both animal and human health. Our investigation was aimed to identify and differentiate between MRSA and methicillin-sensitive Staphylococcus aureus (MSSA) recovered from mastitic milk using MALDI-TOF mass spectrometry compared with phenotypic methods and studying their susceptibility to various antibiotics. Four hundred milk samples from mastitic animals (cows, sheep, goats, and dromedary camels) were investigated. Phenotypic identification of S. aureus was made through MASTASAPH Latex test, STAPH ID 32, and Vitek 2 system. The proteomic characterization of S. aureus was done by MBT. The Kirby Bauer method was accomplished to detect the resistance of S. aureus strains to antibiotics. The results of the MASTASAPH Latex test, revealed that 54 (46%) were recognized as S. aureus. All S. aureus isolates were identified by MBT with a score of more or equal 2.00. Several peaks were identified in the mass of 4590 Da, 4863 Da, and 4938 Da for MSSA and in the mass of 2636 Da and 3009 Da for MRSA. The MSP dendrogram demonstrated that the S. aureus isolates were classified into one group with a distance level of less or equal 400. The percentage of S. aureus resistance against carbenicillin, erythromycin and kanamycin was 94.4%, 38.88%, and 33.33%, respectively. In conclusion, S. aureus bacteria are among the key triggers for mastitis in Saudi Arabia. MBT is reported to be not only the rapid tool to identify S. aureus but also able to discriminate MRSA from MSSA.
“…This technique has allowed the direct identification of bacterial strains of the genus and species, which has represented a cost-effective alternative to classical phenotypic and biochemical assays. In cattle, it is recently used as an alternative tool for identification of mastitis pathogens including S. aureus ( Alharbi et al, 2021 , Barreiro et al, 2017 , El-Ashker et al, 2015 , Nonnemann et al, 2019 , Schmidt et al, 2015 , Shell et al, 2017 ) allowing for more detailed classification of this pathogen ( Liu et al, 2020 ). Besides, it has the potential to become an efficient type of detection method for MRSA identification ( Elbehiry et al, 2016 ).…”
Section: Typing Methods For Bovine
Staphylococcus Aureusmentioning
confidence: 99%
“…This method can directly be applied in large scale in milk samples, which it showed to be simple to implement, reliable and highly reproducible ( Barreiro et al, 2017 , Liu et al, 2020 ). It could analyze samples within a few minutes, and it does not need large amounts of diagnostic material for analysis ( Barreiro et al, 2017 , Shell et al, 2017 ). It is also reported to be able to discriminate MRSA from MSSA ( Alharbi et al, 2021 ).…”
Section: Typing Methods For Bovine
Staphylococcus Aureusmentioning
confidence: 99%
“…It is also reported to be able to discriminate MRSA from MSSA ( Alharbi et al, 2021 ). However, nowadays, MAL-DI-TOF MS is not commonly used in epidemiological studies due to the lack of guidelines for its validity, sensitivity and performance ( Kasela and Malm, 2018 , Shell et al, 2017 ). Furthermore, it is often time-consuming and laborious.…”
Section: Typing Methods For Bovine
Staphylococcus Aureusmentioning
“…Milk samples were used to detect udder bacterial pathogens. Qarter milk samples (10 μL) were cultured to detect udder bacterial pathogens on blood agar with 5% of defibrinated blood with parallel cultivation on selective agar Eosin Methylene Blue Agar (Oxoid, Basingstoke, UK), MacConkey agar (Oxoid, Basingstoke, UK) and Edwards Medium (Oxoid, Basingstoke, UK) (Shell et al 2017). Then, the plates were incubated at 37°C for 24-72 h. We evaluated contamination if the milk samples exhibited three or more different colonies on a culture.…”
Section: Bacterial Isolation and Identification Using Maldi-tof Msmentioning
This study aimed to identify bacterial pathogens in milk samples from dairy cows with subclinical and clinical mastitis as well as to assess the concentrations of oxidant-antioxidant parameters [malondialdehyde (MDA), reduced glutathione (GSH), and total GSH levels] in both blood and milk samples. From a total of 200 dairy cows in 8 farms, 800 quarter milk samples obtained from each udder were tested in the laboratory for the presence of udder pathogens. Cultivated bacteria causing intramammary infection from milk samples were identified by Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF). In addition, from tested animals 60 cows were selected includıng 20 healthy cows that were CMT negative, 20 cows with subclinical mastitis (SM), and 20 cows with clinical mastitis (CM) for detection of MDA, GSH, and total GSH levels in blood and milk samples. Three hundred and eighty (47.5%; 380/800), 300 (37.5%; 300/800), and 120 (15%; 120/800) of milk samples, respectively were CMT positive or SM and CM, and those positives were cows from different farms. We observed that 87.4% (332/380), 25.3% (76/300), and 34.2% (41/120) of cows with CMT positive, CMT negative, and CM had bacterial growth. The most predominantly identified bacteria were Staphylococcus chromogenes (18.7%) obtained mainly from SM and Staphylococcus aureus (16.7%) as the most frequent cause of CM. According to our results, dairy cows with CM had the highest MDA levels, the lowest GSH, and total GSH levels in both blood and milk samples however, high MDA levels and low GSH levels in milk samples with SM were observed. Based on our results, lipid oxidant MDA and antioxidant GSH could be excellent biomarkers of cow's milk for developing inflammation of the mammary gland. In addition, there was no link between nutrition and MDA and GSH levels.
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