scholarly journals Staphylococcus aureus Isolated from Ruminants with Mastitis in Northern Greece Dairy Herds: Genetic Relatedness and Phenotypic and Genotypic Characterization

Toxins ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 176
Author(s):  
Charalampos Kotzamanidis ◽  
George Vafeas ◽  
Virginia Giantzi ◽  
Sofia Anastasiadou ◽  
Stavros Mygdalias ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Genetic Relatedness ◽  
Multiple Correspondence Analysis ◽  
Small Ruminants ◽  
Northern Greece ◽  
Spa Typing ◽  
Diverse Range ◽  
Geographical Regions ◽  
Genotypic Analysis

Staphylococcus aureus is the most common mastitis-related pathogen in dairy cattle, goats, and sheep worldwide. However, the population structure and genomic characteristics of mastitis-associated S. aureus in small ruminants are limited. Furthermore, the genotypic and phenotypic characteristics involved in the pathogenicity of S. aureus have been thoroughly defined, yet their association with the severity of mastitis is not fully established. Here, we performed genotyping by pulsed-field gel electrophoresis (PFGE) and spa analyses to assess the genetic diversity and relatedness of 162 S. aureus strains recovered from clinical mastitis (CM) and subclinical mastitis (SCM) cases from goats, sheep, and bovines. PFGE analysis revealed 108 distinguishable pulsotypes and 3 main clusters that comprised isolates from the three host species, while according to spa typing, 32 different spa types were identified. Genotypic analysis revealed a spreading of genetically related or indistinguishable S. aureus strains among ovine, caprine, and bovine farms of distant geographical regions. In total, 28 different staphylococcal enterotoxin (SE) gene profiles were observed, revealing a diverse range of SE genes among isolates. By evaluating the antimicrobial resistance, we found low phenotypic antimicrobial resistance among all ruminant isolates. We also performed multiple correspondence analysis, which indicated that the presence of the sec gene, biofilm production, and high autoaggregation ability are associated with CM cases.

scholarly journals Massive genome decay and expansion of insertion sequences drive the evolution of a novel host-restricted bacterial pathogen

2020 ◽  
Author(s):  
Gonzalo Yebra ◽  
Andreas F Haag ◽  
Maan M Neamah ◽  
Bryan A Wee ◽  
Emily J Richardson ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Genetic Relatedness ◽  
Bacterial Species ◽  
Small Ruminants ◽  
Bacterial Pathogen ◽  
Insertion Sequences ◽  
Evolutionary Trajectory ◽  
Intergenic Regions ◽  
Genome Decay ◽  
Novel Concept

AbstractBackgroundThe emergence of new bacterial pathogens represents a major threat to public and veterinary health. Staphylococcus aureus is a multi-host bacterial species comprising pathogenic clones with distinct tropisms for human and livestock species. A S. aureus microaerophilic subspecies, Staphylococcus aureus subsp. anaerobius, is responsible for outbreaks of a specific lymphadenitis pathology (Morel’s disease) exclusively found in small ruminants. However, the evolutionary history of S. aureus subsp. anaerobius and its genetic relatedness to S. aureus are unknown.ResultsEvolutionary genomic analyses of clinical S. aureus subsp. anaerobius isolates sampled across 3 decades revealed this clone emerged from a S. aureus progenitor about 1000 years ago (95%CI: 716-1184), before differentiating into two distinct lineages representing African (emerged in 1930 [1907-1951)) and European (1777 [1716-1832]) isolates. S. aureus subsp. anaerobius has undergone limited clonal expansion, with a restricted population size, and an evolutionary rate 10-fold slower than S. aureus. The transition to a highly niche-specific pathogen of small ruminant lymph nodes involved acquisition of a pathogenicity island encoding an effector with ruminant host-specificity, large genomic rearrangements, and the accumulation of at least 205 pseudogenes resulting in a highly fastidious metabolism underpinning its restricted ecological niche. Importantly, acquisition and expansion of ~87 insertion sequences located in conserved intergenic regions provided distinct mechanisms for the control of expression of flanking genes, representing a novel concept of transcriptional regulon.ConclusionsOur findings provide a remarkable example of the evolutionary trajectory of a host-restricted bacterial pathogen that resulted from extensive remodelling of the S. aureus genome.

scholarly journals The Genetic Relatedness and Antimicrobial Resistance Patterns of Mastitis-Causing Staphylococcus aureus Strains Isolated from New Zealand Dairy Cattle

2021 ◽  
Vol 8 (11) ◽  
pp. 287
Author(s):  
Sabrina S. Greening ◽  
Ji Zhang ◽  
Anne C. Midwinter ◽  
David A. Wilkinson ◽  
Scott McDougall ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
New Zealand ◽  
Antimicrobial Resistance ◽  
Genetic Relatedness ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Antimicrobial Usage ◽  
Dry Cow Therapy ◽  
Usage Patterns ◽  
Resistance Patterns

Staphylococcus aureus is one of the leading causes of bovine mastitis worldwide and is a common indication for use of antimicrobials on dairy farms. This study aims to investigate the association between on-farm antimicrobial usage and the antimicrobial resistance (AMR) profiles of mastitis-causing S. aureus. Whole-genome sequencing was performed on 57 S. aureus isolates derived from cows with either clinical or subclinical mastitis from 17 dairy herds in New Zealand. The genetic relatedness between isolates was examined using the core single nucleotide polymorphism alignment whilst AMR and virulence genes were identified in-silico. The association between gene presence-absence and sequence type (ST), antimicrobial susceptibility and dry cow therapy treatment was investigated using Scoary. Altogether, eight STs were identified with 61.4% (35/57) belonging to ST-1. Furthermore, 14 AMR-associated genes and 76 virulence-associated genes were identified, with little genetic diversity between isolates belonging to the same ST. Several genes including merR1 which is thought to play a role in ciprofloxacin-resistance were found to be significantly overrepresented in isolates sampled from herds using ampicillin/cloxacillin dry cow therapy. Overall, the presence of resistance genes remains low and current antimicrobial usage patterns do not appear to be driving AMR in S. aureus associated with bovine mastitis.

scholarly journals Molecular Typing and Antimicrobial Susceptibility Profile of Staphylococcus aureus Isolates Recovered from Bovine Mastitis and Nasal Samples

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2143
Author(s):  
Renata P. Santos ◽  
Fernando N. Souza ◽  
Ana Claudia D. Oliveira ◽  
Antônio F. de Souza Filho ◽  
Juliana Aizawa ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Antimicrobial Susceptibility ◽  
Molecular Typing ◽  
Bovine Mastitis ◽  
Aureus Strain ◽  
Staphylococcal Protein A ◽  
Spa Typing ◽  
Intramammary Infections ◽  
Antimicrobial Susceptibility Profile

In the present study, we aimed to determine the antimicrobial resistance and molecular typing of Staphylococcus aureus recovered from transient and persistent intramammary infections and nares/muzzles in dairy cows. We investigated the antimicrobial resistance of 189 S. aureus strains using a broad antimicrobial susceptibility profile. Furthermore, 107 S. aureus isolates were strain-typed using staphylococcal protein-A (spa) typing. A large proportion of strains exhibited multidrug resistance to antimicrobials, including resistance to critically important antimicrobials, although no methicillin-resistant S. aureus strains were found. Our study did not strengthen the idea that extramammary niches (i.e., nares/muzzles) are an important source of S. aureus for bovine mastitis. A discrepancy in the antimicrobial resistance between S. aureus strains isolated from nares/muzzles and milk samples was observed. Furthermore, S. aureus isolates from transient and persistent intramammary infections (IMIs) did not differ by spa typing, suggesting that the persistence of bovine IMIs was determined by cow factors. Thus, the high level of multidrug-resistant S. aureus found in the two herds, considered together with the predominance of a well udder-adapted S. aureus strain, may contribute to our knowledge of the history of the high prevalence of mastitis caused by S. aureus, which is of great concern for animal and public health.

scholarly journals spa typing and antimicrobial resistance of Staphylococcus aureus from healthy humans, pigs and dogs in Tanzania

2016 ◽  
Vol 10 (02) ◽  
pp. 143-148 ◽  
Author(s):  
Abdul Sekemani Katakweba ◽  
Amandus Pachificus Muhairwa ◽  
Carmen Espinosa-Gongora ◽  
Luca Guardabassi ◽  
Madundo M A Mtambo ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Tetracycline Resistance ◽  
Nasal Carriage ◽  
Opportunistic Pathogen ◽  
Diffusion Method ◽  
Spa Typing ◽  
Healthy Humans ◽  
Mueller Hinton ◽  
Spa Types

Introduction: Staphylococcus aureus is an opportunistic pathogen causing infections in humans and animals. Here we report for the first time the prevalence of nasal carriage, spa typing and antimicrobial resistance of S. aureus in a Tanzanian livestock community. Methodology: Nasal swabs were taken from 100 humans, 100 pigs and 100 dogs in Morogoro Municipal. Each swab was enriched in Mueller Hinton broth with 6.5% NaCl and subcultured on chromogenic agar for S. aureus detection. Presumptive S. aureus colonies were confirmed to the species level by nuc PCR and analysed by spa typing. Antimicrobial susceptibility patterns were determined by disc diffusion method. Results: S. aureus was isolated from 22 % of humans, 4 % of pigs and 11 % of dogs. A total of 21 spa types were identified: 13, 7 and 1 in human, dogs, and pigs, respectively. Three spa types (t314, t223 and t084) were shared between humans and dogs. A novel spa type (t10779) was identified in an isolate recovered from a colonized human. Antimicrobials tested revealed resistance to ampicillin in all isolates, moderate resistances to other antimicrobials with tetracycline resistance being the most frequent. Conclusion: S. aureus carrier frequencies in dogs and humans were within the expected range and low in pigs. The S. aureus spa types circulating in the community were generally not shared by different hosts and majority of types belonged to known clones. Besides ampicillin resistance, moderate levels of antimicrobial resistance were observed irrespective of the host species from which the strains were isolated.

scholarly journals Staphylococcus aureus harbouring egc cluster coding for non-classical enterotoxins, involved in a food poisoning outbreak, Romania, 2012 / Staphylococcus aureus purtător de gene codante pentru enterotoxine non-clasice (cluster egc), implicat într-un focar de toxiinfecţie alimentară, România, 2012

2015 ◽  
Vol 23 (3) ◽  
pp. 285-294 ◽  
Author(s):  
Ileana Luminita Coldea ◽  
Lavinia Zota ◽  
Cristiana Cerasella Dragomirescu ◽  
Brandusa Elena Lixandru ◽  
Elena Carmina Dragulescu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Genetic Relatedness ◽  
Protein A ◽  
Food Poisoning ◽  
Food Handler ◽  
Reference Laboratory ◽  
Food Handlers ◽  
Spa Typing ◽  
Reference Centre ◽  
Laboratory Capacity

Abstract In March 2012, a food poisoning outbreak was reported in a Romanian county, with a total number of 30 children affected. The symptoms (vomiting, diarrhea and abdominal pain), with onset within 1-2 hours after the ingestion of a particular food (milk), suggested a possible staphylococcal aetiology. An outbreak investigation was carried out, in accordance with the national surveillance methodology and 25 samples: stool (n=9), vomit (n=5), nasal swabs (n=9), and milk (n=2) were collected from the affected children, food handlers and suspected food. All isolated strains were sent to the Reference Centre for Staphylococci within the “Cantacuzino” National Institute of Research-Development for Microbiology and Immunology, Bucharest, Romania, for confirmation and further analysis. The aim of this study was to increase the reference laboratory capacity to confirm staphylococcal food poisoning (SFP) outbreak by defining the molecular basis of toxicity of Staphylococcus aureus (S. aureus) isolates and assessing their genetic relatedness. PCR methods have been used to detect 14 enterotoxin genes and the expression of some of these genes was proved by using a reverse transcription real-time PCR. Pulsed-field gel electrophoresis (PFGE) and Staphylococcus protein A coding gene sequence typing (spa typing) have been used to track the origin of the S. aureus contamination and to confirm the food poisoning outbreak. Two enterotoxigenic S. aureus strains isolated from milk, twelve isolated from patients and two from food handlers were of the same spa- type (t902) and revealed an indistinguishable SmaI macrorestriction pattern after a PFGE analysis. All these strains harboured the same toxin genes profile, namely the enterotoxin gene cluster (egc), which strongly supports the evidence that the milk was the incriminated food vehicle of the outbreak and a food-handler was the most likely source of the staphylococcal food poisoning (SFP) incident.

scholarly journals Biofilm Production Ability, Virulence and Antimicrobial Resistance Genes in Staphylococcus aureus from Various Veterinary Hospitals

Pathogens ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 264
Author(s):  
Lin Chen ◽  
Zi-Yun Tang ◽  
Shi-Yun Cui ◽  
Zhen-Bao Ma ◽  
Hua Deng ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Protein A ◽  
Production Capacity ◽  
Spa Typing ◽  
Airborne Dust ◽  
Biofilm Production ◽  
Antimicrobial Resistance Genes ◽  
Different Origins

Staphylococcus aureus (S. aureus) is one of the most clinically important zoonotic pathogens, but an understanding of the prevalence, biofilm formulation ability, virulence, and antimicrobial resistance genes of S. aureus from veterinary hospitals is lacking. By characterizing S. aureus in different origins of veterinary hospitals in Guangzhou, China, in 2019, we identified with the presence of S. aureus in pets (17.1%), veterinarians (31.7%), airborne dust (19.1%), environmental surfaces (4.3%), and medical device surfaces (10.8%). Multilocus sequence typing (MLST) and Staphylococcus protein A (spa) typing analyses demonstrated methicillin-sensitive S. aureus (MSSA) ST398-t571, MSSA ST188-t189, and methicillin-resistant S. aureus (MRSA) ST59-t437 were the most prevalent lineage. S. aureus with similar pulsed-field gel electrophoresis (PFGE) types distributed widely in different kinds of samples. The crystal violet straining assays revealed 100% (3/3) of MRSA ST59 and 81.8% (9/11) of MSSA ST188 showed strong biofilm formulation ability, whereas other STs (ST1, ST5, ST7, ST15, ST88, ST398, ST3154 and ST5353) showed weak biofilm production ability. Polymerase chain reaction (PCR) confirmed the most prevalent leucocidin, staphylococcal enterotoxins, ica operon, and adhesion genes were lukD-lukE (49.0%), sec-sel (15.7%), icaA-icaB-icaC-icaR (100.0%), and fnbB-cidA-fib-ebps-eno (100.0%), respectively. Our study showed that the isolates with strong biofilm production ability had a higher prevalence in clfA, clfB, fnbA and sdrC genes compared to the isolates with weak biofilm production ability. Furthermore, 2 ST1-MRSA isolates with tst gene and 1 ST88-MSSA isolate with lukS/F-PV gene were detected. In conclusion, the clonal dissemination of S. aureus of different origins in veterinary hospitals may have occurred; the biofilm production capacity of S. aureus is strongly correlated with ST types; some adhesion genes such as clfA, clfB, fnbA, and sdrC may pose an influence on biofilm production ability and the emergence of lukS/F-PV and tst genes in S. aureus from veterinary hospitals should raise our vigilance.

scholarly journals Spa typing of Staphylococcus aureus Isolated from Clinical Specimens from Outpatients in Iraq

2021 ◽  
Vol 70 (1) ◽  
pp. 79-85
Author(s):  
KHAIRALLAH A.S. MOHAMMED ◽  
ZAHRAA H. ABDULKAREEM ◽  
AYOOB R. ALZAALAN ◽  
AMEL K. YAQOOB
Keyword(s):  
Staphylococcus Aureus ◽  
Genetic Relatedness ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Spa Typing ◽  
Disk Diffusion Method ◽  
Vitek 2 ◽  
Species Specific ◽  
Spa Types

Methicillin-resistant Staphylococcus aureus (MRSA) is notorious as a hospital superbug and a problematic pathogen among communities. The incidence of MRSA has substantially increased over time in Iraq. The aim of this study was to determine the prevalence and spa types of MRSA isolates from outpatients or patients upon admission into hospitals. Various biochemical tests identified S. aureus isolates, and then this identification was confirmed by PCR using species-specific 16S rRNA primer pairs. Antibiotic susceptibility was determined against methicillin, oxacillin, and vancomycin using the disk diffusion method. Vancomycin MIC was detected by VITEK 2 compact system. All the identified isolates were screened for the presence of mecA and lukS-PV-lukF-PV genes; 36 of them were subjected to spa typing-based PCR. Out of 290 clinical samples, 65 (22.4%) were S. aureus, of which 62 (95.4%) strains were resistant to oxacillin and methicillin. Except for two isolates, all MRSA isolates were mecA positive. One of the three MSSA isolates was mecA positive. Five strains were resistant to vancomycin. Fourteen (21.5%) isolates were positive for the presence of lukS-PV-lukF-PV genes. Spa typing of 36 S. aureus isolates revealed eleven different spa types, t304 (30.3%), t307 (19.4%), t346 (8.3%), t044 (8.3%), t15595 (8.3%), t386 (5.5%), t5475 (5.5%), t17928 (2.8%), t14870 (2.8%), t021 (2.8%), and t024 (2.8%). These findings could be useful for assessing the genetic relatedness of strains in the region for epidemiological and monitoring purposes, which would be essential to limiting the spread of MRSA.

scholarly journals Distribution and Clonal Diversity of Staphylococcus aureus and Other Staphylococci in Surface Waters: Detection of ST425-t742 and ST130-t843 mecC-Positive MRSA Strains

Antibiotics ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1416
Author(s):  
Vanessa Silva ◽  
Eugénia Ferreira ◽  
Vera Manageiro ◽  
Lígia Reis ◽  
María Teresa Tejedor-Junco ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Multidrug Resistance ◽  
Antimicrobial Resistance ◽  
Virulence Factors ◽  
Resistance Genes ◽  
Surface Waters ◽  
Water Samples ◽  
Coagulase Negative Staphylococci ◽  
Spa Typing ◽  
Antimicrobial Resistance Genes

Natural aquatic environments represent one of the most important vehicles of bacterial dissemination. Therefore, we aimed to isolate staphylococci from surface waters and to investigate the presence of antimicrobial resistance genes and virulence factors as well as the genetic lineages of all Staphylococcus aureus isolates. Staphylococci were recovered from water samples collected from 78 surface waters, including rivers, streams, irrigation ditches, dams, lakes, and fountains. The presence of antimicrobial resistance genes and virulence factors was investigated by PCR. Multilocus sequence typing and spa-typing were performed in all S. aureus isolates. From the 78 water samples, 33 S. aureus, one S. pseudintermedius, and 51 coagulase-negative staphylococci (CoNS) were identified. Among the S. aureus isolates, four MRSA were identified, and all harbored the mecC gene. Fourteen S. aureus were susceptible to all antimicrobials tested and the remaining showed resistance to penicillin, erythromycin and/or tetracycline encoded by the blaZ, ermT, msr(A/B), tetL, and vgaA genes. Regarding the clonal lineages, one mecC-MRSA isolate belonged to spa-type t843 and sequence type (ST) 130 and the other three to t742 and ST425. The remaining S. aureus were ascribed 14 spa-types and 17 sequence types. Eleven species of CoNS were isolated: S. sciuri, S. lentus, S. xylosus, S. epidermidis, S. cohnii spp. urealyticus, S. vitulinus, S. caprae, S. carnosus spp. Carnosus, S. equorum, S. simulans, and S. succinus. Thirteen CoNS isolates had a multidrug resistance profile and carried the following genes: mecA, msr(A/B), mph(C), aph(3′)-IIIa, aac(6′)-Ie–aph(2′’)-Ia, dfrA, fusB, catpC221, and tetK. A high diversity of staphylococci was isolated from surface waters including mecCMRSA strains and isolates presenting multidrug-resistance profiles. Studies on the prevalence of antibiotic-resistant staphylococci in surface waters are still very scarce but extremely important to estimate the contribution of the aquatic environment in the spread of these bacteria.

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