Molecular and epidemiological characterization of Staphylococcus aureus causing bovine mastitis in China.

Staphylococcus aureus is one of the most prevalent pathogens in bovine mastitis, which leads to substantial financial losses for the dairy industry. In this study, S. aureus (n = 72) was isolated from 18 dairy farms in 15 provinces across China in 2021. The identification of these isolates at the species level was achieved by employing 16S rRNA sequencing. An isothermal amplification method for auxiliary detection of S. aureus was established, which can be employed not only for laboratory detection but also for point-of-care testing (POCT). Molecular characteristics of S. aureus mastitis in Chinese dairy cows were determined through MLST and spa typing. Finally, methicillin-resistant Staphylococcus aureus (MRSA) and MRSA resistance genes were detected using MIC and PCR amplification techniques. 72 isolates were identified as 12 sequence types (STs) and 7 clonal complexes (CC). ST1/CC1 was the dominant prevalent accounting for 33.3 % of the total, and exhibiting a wide distribution range. In terms of spa types, t114 was the dominant type, accounting for 31.9 % of the total, followed by t529 as the second major type. Four S. aureus strains were classified as MRSA according to their levels of oxacillin resistance (MIC ≥4 μg/mL). Among these four MRSA strains, one of them was found to be mecA positive. However, the presence of drug-resistance genes mecA and mecC was not detected in the remaining three MRSA strains, indicating the possible existence of new resistance genes. Our study investigated the prevalence of S. aureus mastitis in dairy cows in China, while also examined the molecular characteristics and MRSA strains. This information will help with the clinical monitoring, prevention, and control of S. aureus mastitis in dairy cattle.

Zhu L ，Lai Y ，Li X ，Ma H ，Gong F ，Sun X ，Cao A ，Jiang T ，Han Y ，Pan Z ... -  《-》

Antimicrobial resistance, virulence gene profile and molecular typing of Staphylococcus aureus isolates from dairy cows in Xinjiang Province, northwest China.

Staphylococcus aureus (SA) is a major pathogen causing dairy cow mastitis and endometritis. Recently, animal-derived SA strains pose a serious public-health threat. However, little is known about antimicrobial resistance and virulence factors of SA isolated from dairy cows in Xinjiang, China. In this study, antimicrobial resistance, virulence gene profiles and genotypes of SA from clinical mastitis and endometritis in dairy cows were investigated. A total of 337 clinical samples (186 milk samples from clinical mastitis cases and 151 endometritis swab samples) were collected from 15 large-scale dairy farms and were screened for SA. All SA isolates were subjected to antimicrobial susceptibility testing, detection of virulence genes and molecular typing. A total of 155 SA strains were isolated; 22 (14.2%) were methicillin-resistant S. aureus (MRSA). Resistance of MRSA isolates was significantly higher than that of methicillin-susceptible S. aureus (MSSA). The percentage of virulence genes varied between MSSA and MRSA. The strains could be divided into two SCCmec types (I and IVa), three agr types (I, II and III) and four spa types (t779, t2883, t13751 and t1939). MLST identified 14 sequence types, among which ST1 and ST9 had relatively high detection rates. These findings revealed that ST9-t1939-agrI was the main genotype of MSSA, whilst ST1-SCCmecI-t1939-agrI was the main genotype of MRSA from dairy cows. More significantly, a novel ST (STX) was identified for the first time. The majority of SA strains from dairy cows were multidrug-resistant and carried multiple virulence genes, posing a potential public-health risk.

Dan M ，Yehui W ，Qingling M ，Jun Q ，Xingxing Z ，Shuai M ，Kuojun C ，Jinsheng Z ，Zibing C ，Zaichao Z ，Xuepeng C ... -  《-》

Molecular Characteristics of Staphylococcus aureus Causing Bovine Mastitis between 2014 and 2015.

Li T ，Lu H ，Wang X ，Gao Q ，Dai Y ，Shang J ，Li M ... -  《Frontiers in Cellular and Infection Microbiology》

Bovine mastitis Staphylococcus aureus: antibiotic susceptibility profile, resistance genes and molecular typing of methicillin-resistant and methicillin-sensitive strains in China.

The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infection in dairy animals is of great concern for livestock and public health. The aim of present study was to detect new trends of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) towards antibiotic susceptibility, resistance genes and molecular typing by methods of disc diffusion, multiplex PCR assay and multilocus sequence typing (MLST). A total of 219 S. aureus strains were isolated from bovine mastitis cases from six provinces of China, including 34 MRSA strains. The results revealed that more than 70% isolated strains showed resistance to various antibiotics, and multiple-drugs resistance to more than five categories of antibiotics was found more common. The ermC was the most prevalent resistance gene, followed by other genes; however, ermA was the least frequently detected gene. Twenty-eight mecA-negative MRSA and six mecA-positive MRSA strains were detected, and in which three strains were ST97-MRSA-IV, others were ST965-MRSA-IV, ST6-MRSA-IV and ST9-MRSA-SCCmec-NT. The mecA-negative MRSA strains were found resistant to most of the antibiotics, and harbored aac(6')/aph(2''), aph(3')-III and tetM genes higher than MSSA strains. The resistance to most of the antibiotics was significantly higher in MRSA than in MSSA strains. The MLST profiles showed that these strains mainly belonged to CC5, CC398, CC121 and CC50 lineage, especially within ST97 and ST398, while some novel sequence types (ST2154, ST2165 and ST2166) were identified and deposited in the MLST database. This indicates that the resistance of S. aureus is becoming more complicated by changes in multi-drug resistance mechanism and appearance of mecA-negative MRSA isolates, and importantly, MRSA-IV strains in different MLST types are emerging.

Wang D ，Wang Z ，Yan Z ，Wu J ，Ali T ，Li J ，Lv Y ，Han B ... -  《-》

Molecular characterization and clonal diversity of methicillin-susceptible Staphylococcus aureus in milk of cows with mastitis in Brazil.

Mastitis is an important disease for the dairy industry worldwide, causing economic losses and reducing milk quality and production. Staphylococcus aureus is a worldwide agent of this intramammary infection, which also causes foodborne diseases. The objective of this study was to determine the frequency of methicillin-susceptible Staphylococcus aureus (MSSA) isolates in milk of mastitis cows in Brazil and to analyze the genetic lineages and the content of antimicrobial resistance genes and virulence factors among these isolates. Fifty-six MSSA isolates were recovered from 1,484 milk samples (positive for the California mastitis test) of 518 cows from 11 different farms in Brazil (representing 51% of total Staph. aureus obtained), and they were further characterized. Methicillin-susceptible Staphylococcus aureus were isolated from 3.7% of California mastitis test-positive tested milk samples and from 6.2% of tested mastitic cows. Methicillin-susceptible Staphylococcus aureus isolates were characterized by spa typing, agr typing, and multilocus sequence typing, and resistance and virulence traits were investigated by PCR. Seven spa types were identified among MSSA (% of isolates): t127 (44.6), t605 (37.5), t002, t1784, t2066 (1.8), and 2 new ones: t10856 (10.7) and t10852 (1.8). Five distinct sequence types (ST) were detected (% of isolates): ST1 (46.4), ST126 (37.5), ST133 (10.7), ST5 (3.6), and a novel ST registered as ST2493 (1.8). Resistances were detected for streptomycin, chloramphenicol, and tetracycline. One strain contained the chloramphenicol resistance gene (fexA; included within transposon Tn558) and 3 strains contained the tetracycline resistance gene [tet(K)]. Methicillin-susceptible Staphylococcus aureus strains were susceptible to most of the antibiotics studied and lacked the virulence genes of Panton-Valentine leukocidin (lukF/S-PV), toxic shock syndrome toxin 1 (tst), exfoliative toxin A (eta), and exfoliative toxin B (etb), as well as the genes of the immune evasion cluster. Methicillin-susceptible Staphylococcus aureus isolates were detected in a relatively low proportion of cows with mastitis (6.2%) and recovered isolates presented high diversity of genetic lineages, with CC1 and CC126 the predominant clonal complexes, and CC133 also being detected. Larger epidemiological studies with molecular characterization of isolates are required to deepen the knowledge on the circulating genetic lineages among the cow population with mastitis.

Silva NCC ，Guimarães FF ，Manzi MP ，Budri PE ，Gómez-Sanz E ，Benito D ，Langoni H ，Rall VLM ，Torres C ... -  《-》