Next Article in Journal
Mortality of Pandrug-Resistant Klebsiella pneumoniae Bloodstream Infections in Critically Ill Patients: A Retrospective Cohort of 115 Episodes
Next Article in Special Issue
Impact of On-Farm Interventions against CTX-Resistant Escherichia coli on the Contamination of Carcasses before and during an Experimental Slaughter
Previous Article in Journal
Antimicrobial Resistance of Non-O157 Shiga Toxin-Producing Escherichia coli Isolated from Humans and Domestic Animals
Previous Article in Special Issue
Molecular Epidemiology of Antibiotic-Resistant Escherichia coli from Farm-to-Fork in Intensive Poultry Production in KwaZulu-Natal, South Africa
Article

Prevalence and Characterization of Coagulase Positive Staphylococci from Food Products and Human Specimens in Egypt

1
Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, University of Sadat City, Sadat City 32897, Egypt
2
Department of Animal Medicine and Infectious Diseases, Faculty of Veterinary Medicine, University of Sadat City, Sadat City 32897, Egypt
3
Division of Public Health, Veterinary Hospital, Khanka, Benha 13736, Egypt
4
Department of Husbandry and Animal Wealth Development, Faculty of Veterinary Medicine, University of Sadat City, Sadat City 32897, Egypt
5
Department of Food Hygiene and Control, Faculty of Veterinary Medicine, University of Sadat City, Sadat City 32897, Egypt
6
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh
7
Clinical Laboratory Sciences Department, Turabah University College, Taif University, Taif 21995, Saudi Arabia
8
Department of Biochemistry, Faculty of Veterinary Medicine, Benha University, Toukh 13736, Egypt
9
Department of Pharmacology and Therapeutics, Faculty of Veterinary Medicine, Damanhour University, Damanhour 22511, Egypt
10
Center of Excellence for Screening of Environmental Contaminants, Benha University, Toukh 13736, Egypt
11
Department of Food Hygiene and Control, Faculty of Veterinary Medicine, Benha University, Toukh 13736, Egypt
12
Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Benha University, Toukh 13736, Egypt
*
Author to whom correspondence should be addressed.
Received: 7 November 2020 / Revised: 11 January 2021 / Accepted: 12 January 2021 / Published: 14 January 2021
(This article belongs to the Special Issue Antimicrobial Resistance: From Farm to Fork)

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) strains have veterinary and public health importance as they are responsible for a wide range of difficult to treat infections and food poisoning. Two hundred samples (50 samples each of minced meat, beef luncheon, Karish cheese, and human samples (pus swab from open wounds)) were cultured, and MRSA strains were identified using disk diffusion tests and mecA gene-based PCR. A total of 35% (70/200) of the examined samples were confirmed as coagulase-positive S. aureus in minced meat (46%), beef luncheon (44%), Karish cheese (44%), and human samples (22%). The MRSA strains showed resistance to amoxicillin (91.4%), penicillin (97.1%), cefoxitin (85.7%), cephradine (82.9%), tetracycline (57.2%), and erythromycin (52.8%). More than half of the tested S. aureus isolates harbored the mecA gene. The sequence analysis of the mecA gene from the minced meat, Karish cheese, and human samples revealed high genetic similarities between the S. aureus isolates from these sources. In conclusion, our findings indicate a risk for the transmission of the mecA gene of S. aureus across the food chain between humans and animal food products. Further studies should focus on finding additional epidemiological aspects of the MRSA strains in food chain.
Keywords: Staphylococcus aureus; antimicrobial resistance; food safety; food products; mecA gene Staphylococcus aureus; antimicrobial resistance; food safety; food products; mecA gene

1. Introduction

Staphylococcus aureus is recognized as a zoonotic pathogen of public health concern, producing serious illness in humans, subclinical mastitis in cows, and outbreaks of food poisoning [1,2]. Globally, S. aureus causes widely distributed opportunistic, nosocomial, and population-related diseases [3]. A recent report stated that S. aureus strains can spread to different geographical regions through food and milk products [4]. According to a surveillance study in India, a lack of hygienic procedures during processing and post-processing handling of food products, particularly those of animal origins such as eggs, meat, and milk, facilitated microbial contamination [5].
S. aureus has developed several virulence factors that aid in pathogenesis and udder infection, such as evasion and adhesion within the host and resistance to the immunologic responses [6,7].
Production of coagulase is a crucial phenotypic feature and the major determinant factor for identifying S. aureus strains. Moreover, the variability at the 3′ end coding region of the coa gene has been used for genotyping of S. aureus strains from humans and animals [8].
Methicillin-resistant S. aureus (MRSA) has frequently been isolated from livestock, and the possibility of zoonotic spread to humans has been recognized. For example, the MRSA ST398, a model clone of livestock-associated MRSA (LA-MRSA), has been found in human population that are in frequent contact with animals such as farmers or veterinarians [9]. Outbreaks of the MRSA infection have been reported in the USA, Europe, and Egypt [10,11].
Several studies have highlighted the evolution of resistance to numerous antimicrobial drugs in S. aureus strains obtained from bovines [12]. S. aureus has also been implicated in food poisoning outbreaks, healthcare, and community-associated infection, particularly MRSA strains that exhibit multidrug resistance (MDR) [11,13,14]. The widespread predominance of MDR-MRSA strains poses a significant risk to public health, and invasion of MDR strains could theoretically transfer resistance genes to humans [13].
In fact, certain bacterial strains of the genus Staphylococcus that have developed resistance to various antibiotics carry the mecA gene that encodes penicillin-binding protein 2a (PBP2a) and facilitate resistance to methicillin and other β-lactam antibiotics, rendering them a global public health issue [14]. Molecular identification using PCR is the gold standard approach for identifying the mecA gene between MRSA strains [15]. However, a special mecA homolog gene was detected in some S. aureus strains, primarily the mecALGA251 (mecC), and this divergent mecA gene counterpart with 70% homology to the mecA gene has been designated as mecC [16].
Karish cheese is a common local and popular type of white, soft, lactic cheese made from raw milk or laban rayeb (raw cottage cheese with salt) which is not exposed to any heat treatment. Additionally, minced meat and beef luncheon are very common and popular raw meat products consumed by the Egyptian people which require no or low heat treatment during cooking. The frequent consumption of such food products of animal origin in developing countries such as Egypt without health oversight can result in the spread of certain bacterial infections such as S. aureus. Therefore, this study aimed to investigate the prevalence and antibiogram profile of S. aureus isolates in animal food products such as minced meat, beef luncheon, and Karish cheese, as well as in human samples. Moreover, partial sequencing of the mecA gene of three MRSA strains has been performed to compare the identity of the mecA gene with related global strains as well as to highlight the possibility of transmitting the antibiotic resistance genes to humans through animal food products and its potential public health significance.

2. Materials and Methods

2.1. Ethical Statement

The medical staff of Al-Qaluobyia Hospital collected the human samples used in this study following the rules and regulations of the Egyptian Ministry of Health under full medical supervision. Written consent has been obtained from the patients before using the specimens. Food samples (minced meet, beef luncheon, and Karish cheese) were collected randomly from local markets in the Al-Qaluobyia Governorate.

2.2. Study Area

The study was conducted in the Al-Qaluobyia governorate in Lower Egypt, about 35.26 km north of Cairo in the Nile Delta region (30.41 N: _N 31.21_E at an altitude of 9.0 m). The study area (1124 km2) had a rapid urbanization over the last three decades. According to inhabitance estimates, in 2018, most residents in the governorate lived in rural areas, with an urbanization rate of only 44.7%. Out of 5,703,000 people located in the governorate, 2,509,000 people lived in rural areas and 3,194,000 lived in urban areas [17]. In addition, the Qaluobyia governorate is the leader in poultry and egg production and food packaging and processing in Egypt.

2.3. Collection of Samples and Preparation

A total of 200 samples (50 samples each of minced meat, beef luncheon, Karish cheese, and human samples (pus swab from open wound)) were collected from December 2018 to December 2019. The food products samples were collected from different areas in the same governorate from shops (Al-Qalyubia governorate). Samples were collected under aseptic condition. The human pus samples were collected from hospitals in the same area by medical staff under the supervision of a physician. The samples were collected in accordance with Americana Public Health Association (APHA), 1992 [18] for the food samples and in accordance with Rice et al., 2016 for human samples [19]. The samples were transferred to the laboratory in sterile plastic bags under hygienic and aseptic cooled conditions for bacteriological examination. One gram of minced meat, beef luncheon, and Karish cheese was mashed with 9 mL of nutrient broth and incubated for 18 h at 37 °C as described previously [20]. The extracted pus swab from humans was grown in nutrient broth and incubated for 18 h at 37 °C under aseptic conditions in accordance with Rice et al. [19].

2.4. Phenotypic Isolation and Identification of Staphylococcus aureus

The processed samples were incubated for 1-2 days at 37 °C in Baird–Parker agar (Oxoid Ltd., Basingstoke, UK) and 10% sheep blood agar (Oxoid Ltd., UK). Staphylococci were confirmed through Gram staining, catalase, and coagulase test [20]. Virulence activities such as hemolytic activity and DNase agar testing were performed in accordance with Murray et al. [21]. Biofilm activity on Congo red medium was tested in accordance with Freeman et al. [22].

2.5. Antibiogram Profile of Staphylococcus aureus Isolates

The antibiogram profile of 70 S. aureus isolates was tested in vitro using the Kirby–Bauer disk diffusion method. A suspension of the bacteria was adjusted to a 0.5 McFarland suspension. Twelve antimicrobials disks (Oxoid Ltd., UK) of different antibiotic groups were used to detect the antimicrobial susceptibility and resistance patterns of these isolates. The following antibiotics were used: 100 IU penicillin (P), 25 µg amoxicillin (AX), 30 µg cefoxitin (FOX), 30 µg vancomycin (VA), 10 µg gentamicin (CN), 15 µg erythromycin (E), 30 µg tetracycline (TE), 5 µg ciprofloxacin (CIP), 10 µg norfloxacin (NOR), 30 µg cephradine (CE), chloramphenicol (C), and 1.25/23.75 µg trimethoprim-sulfamethoxazole (SXT). The results were interpreted as resistant, intermediate, or susceptible according to the inhibitory zone as described by the Clinical and Laboratory Standards Institute (CLSI) [23]. The S. aureus ATCC 25923 was used as a quality control organism in antimicrobial susceptibility determination.

2.6. Molecular Characterization of Staphylococcus aureus Isolates

DNA was extracted using the QIAamp DNA Mini Kit (Cat. No. 51304, Qiagen, Hilden, Germany). The list of primers used to amplify the coa and mecA genes of S. aureus isolates is provided in Table 1. For PCR, a reaction mixture of 50 µL volume was prepared, containing 25 µL of PCR Master Mix (Cat. No. 201445, Qiagen, Germany), 1 µL (10 pmol/µL) of each primer, 6 µL target DNA, and the remaining volume needed to reach 50 µL was adjusted with deionized water. The reaction was conducted in an Applied Biosystem 2720 thermal cycler (Applied Biosystems, Foster, CA, USA). The following thermal profile was used: Initial denaturation at 94 °C for 5 min; 40 cycles each consisting of denaturation at 94 °C for 30 sec, annealing at 55 °C (coa gene) or 57 °C (mecA gene) for 45 sec and extension at 72 °C for 30 sec; and finally extension at 72 °C for 5 min. After completing the run, PCR products (15 µL) were subjected to agarose gel electrophoresis (1.5%) and visualized under UV light in a gel documentation system.
Table 1. PCR primers and conditions for coa and mecA gene amplification.
Table 1. PCR primers and conditions for coa and mecA gene amplification.
TargetPrimersSequences (5′ to 3′)Amplicon Size (bp)Annealing TemperatureReferences
mecAForwardAGA AGA TGG TAT GTG GAA GTT AG58357 °C[24]
ReverseATG TAT GTG CGA TTG TAT TGC
CoaForwardACC ACA AGG TAC TGA ATC AAC G600–100055 °C[25]
ReverseTGC TTT CGA TTG TTC GAT GC

2.7. Sequencing and Analysis of Staphylococcus aureus mecA Gene

The PCR products were purified using GeneJET PCR Purification Kit (Cat no. K0701, ThermoFisher Scientific, USA) and sequenced from a commercial laboratory (GATC Biotech Company, Germany) in an ABI 3730xl DNA sequencer using both forward and reverse primers. To identify homologies in the nucleotide and amino acid sequences, the isolated strains were compared with S. aureus strains available in the GenBank using BLAST 2.0 and PSI- BLAST search databases (National Center for Biotechnology Information NCBI at http://0-www.ncbi.nlm.nih.gov.brum.beds.ac.uk/), respectively. The nucleotide sequence comparison and multiple alignments with reference strains and the deduced amino acid sequence analysis were conducted as described previously [26] using the BioEdit sequence alignment editor, CLUSTALX software for multiple sequence alignment. ClustalW software for multiple sequence alignment [27]. ClustalV [28] and MegAlign (DNASTAR, Lasergene®, Version 7.1.0, Madison, WI, USA) [29] Phylogenetic trees were constructed using MegAlign using the neighbor-joining method based on ClustalW. Bootstrapping values were estimated using a random seeding value of 111 [27]. ClustalV was applied when end gaps were faced. Sequence deviation and identity percent were calculated using MegAlign and the structural character of our protein sequence was identified using Protean (DNASTAR, Lasergene®, Version 7.1.0., Madison, WI, USA).

2.8. Statistical Analysis

To visualize the multidrug resistance profile of MRSA strains, an UpSetR plot was prepared using an online platform (https://gehlenborglab.shinyapps.io/upsetr/).

3. Results

3.1. Prevalence of Coagulase-positive Staphylococci Isolated from Food Products and Human Samples

At first, we investigated the prevalence of coagulase-positive Staphylococci (CPS) in animal food products and human specimens. To this end, we collected 200 samples of minced meat, beef luncheon, Karish cheese, and human pus swabs. Out of 200 samples, 123 samples showed phenotypically positive growth on Baird–Parker medium. The prevalence of CPS was 24 (48%), 42 (84%), 38 (76%), and 19 (38%) in minced meat, beef luncheon, Karish cheese, and human swabs samples, respectively (Table 2).

3.2. Biochemical Activity and Virulence Factors of the Coagulase-positive Staphylococci (CPS) Isolates

Next, we tested the biochemical and enzymatic activity of the CPS isolates. Out of 123 isolates, 70 strains were confirmed as coagulase-positive S. aureus by coagulase test. The hemolysis activity on blood agar media was found in 23 (46%), 22 (44%), 22 (44%), and 11 (22%) CPS isolates from minced meat, beef luncheon, Karish cheese, and human samples, respectively. In addition, the DNase activity was found in 24 (48%), 42 (84%), 38 (76%), and 19 (38%) CPS isolates detected in minced meat, beef luncheon, Karish cheese, and human samples, respectively. The lecithinase activity was noticed in 21 (42%), 2 (4%), and 1 (2%) CPS isolates from minced meat, beef luncheon, and human samples, respectively, while no isolates from Karish cheese exhibited lecithinase activity. Biofilm activity on Congo red medium was observed in 19 (38%), 37 (74%), 32 (64%), and 19 (38%) CPS isolates from minced meat, beef luncheon, Karish cheese, and human samples, respectively. Collectively, the CPS isolates detected in animal food products, and human samples carried potential virulence factors.

3.3. Antimicrobial Resistance of the Coagulase-positive Staphylococci (CPS) Isolates

We performed the antimicrobial resistance analysis of the purified S. aureus isolates obtained from animal food products and human samples using the disc diffusion technique. A total of 12 antibiotics belonging to eight antimicrobial classes were used in the analysis (Table 3). The antibiotic sensitivity test of the 70 strains of S. aureus revealed a relatively high antibiotic resistance property of these isolates. The S. aureus isolates showing resistance to antibiotics were as follows: penicillin (97.1%), amoxicillin (91.4%), cefoxitin (85.7%), cephradine (82.9%), tetracycline (57.2%), and erythromycin (52.8%). On the other hand, a relatively higher sensitivity of the S. aureus isolates was detected to vancomycin (84.3%), chloramphenicol (70%), norfloxacin (55.7%), trimethoprim-sulfamethoxazole (50%), gentamicin (44.3%), and ciprofloxacin (42.8%) (Table 3). Collectively, the antimicrobial sensitivity assay revealed the presence of a large number of antibiotic-resistant S. aureus isolates from animal food products.

3.4. Multidrug Resistance (MDR) Profiles of Methicillin-Resistant Staphylococcus aureus (MRSA) Strains from Animal Food Products and Human Samples

The multidrug resistance (MDR) profiles of the 70 S. aureus isolates were analyzed using 12 different antibiotics belonging to eight antimicrobial classes. The MDR profiles of the S. aureus isolates are presented in Figure 1 and Supplementary Table S1. Of note, two of the S. aureus isolates showed resistance to all 12 antibiotics belonging to eight antimicrobial classes. Seven isolates showed resistance of 11 antibiotics belonging to seven antimicrobial classes. A total of 12 S. aureus isolates exhibited resistance to 8–10 different antibiotics belonging to six different antibiotics classes. Another 12 S. aureus isolates showed resistance to 6–9 antibiotics belonging to five antimicrobial classes. In addition, MDR to 4–8 antibiotics belonging to four antimicrobial classes was observed in 12 S. aureus isolates. MDR to three and two antibiotic classes was found in five and 10 S. aureus isolates, respectively. Taken together, most of the S. aureus isolates from animal food products were multidrug-resistant.

3.5. Molecular Characterization of the Staphylococcus aureus Strains

For molecular characterization of the S. aureus strains, we first tested the presence of the coa gene in the S. aureus strains isolated from the animal food products and human pus samples. To this end, we randomly selected nine S. aureus strains isolated from minced meat, beef luncheon, Karish cheese, and human swabs samples and amplified the coa gene by PCR. The PCR method successfully amplified a 600–1000 nucleotide fragment of the coa gene of S. aureus strains in all samples (Supplementary Figure S1). Two isolates of minced meat, beef luncheon, and Karish cheese were positive for the coa gene. In addition, the coa gene was detected in three isolates obtained from human pus samples.
Next, we tested the presence of the mecA gene in the S. aureus strains isolated from minced meat, beef luncheon, Karish cheese, and human pus samples. For this, we selected 19 S. aureus strains that exhibited strong reactions in biochemical tests and virulence factors including coagulase test, DNase-, hemolysis on blood agar, and Biofilm activities. The PCR technique amplified a 583-nucleotide fragment of the mecA gene of S. aureus strains in 11 isolates (Supplementary Figure S2). The mecA gene was detected in two isolates each of minced meat, beef luncheon, and Karish cheese. Five isolates from human pus samples were also positive for the mecA gene.
Then, we tested the genetic diversity of the mecA gene in the S. aureus strains isolated from animal food products and human samples. We sequenced the partial mecA gene of three S. aureus strains to analyze the genetic similarities between the S. aureus from human and animal food products. The three selected strains of S. aureus were obtained from three different sources (human, minced meat, and Karish cheese), were multidrug-resistant strains, and had the same properties as other strains from the same source in terms of morphology and biochemical and virulence activities. The mecA partial gene sequences from human, minced meat, and Karish cheese samples were submitted in the GenBank with accession numbers MT185692, MT211620, and MT211621, respectively. The mecA gene sequences of our three S. aureus strains showed high nucleotide similarity (99.79 to 100%) with mecA gene sequences of S. aureus reported from different countries. The phylogenetic tree also clustered these isolates with the S. aureus isolates obtained from different geographical locations (Figure 2). For example, we detected nearly identical sequences of the mecA gene detected in three S. aureus strains with the mecA genes of S. aureus strains detected in human samples from the USA (HL20709; ER03364.3, and ER03868.3), three isolates of pigs and human origin from China (SH6P021P, LTH-uvas, and ZY05), one human specimen from Pakistan (RJ1267), and one human urine sample from Japan (SC955). The protein identity and similarity between our isolates and isolates from other countries are illustrated in the phylogenetic tree (Figure 3). The result showed that nearly identical protein sequences of the mecA gene of the study strains have been found in different geographical areas including two isolates from China (Guangzhou-SAu071 and WH9628) obtained from human bile and sputum specimens and with one isolate each from the USA and South Korea obtained from human specimens (ER03364.3 and HL20709). Additionally, the phylogenetic tree clustered our isolates with S. aureus isolates from several countries. For example, we observed high amino acid sequence similarities of the mecA gene of our study with one isolate from a pig in Pakistan (LTH-uVas) and two isolates from human sputum and blood specimens in China (RJ1267 and ZY05). Furthermore, three human isolates from three different countries showed high genetic similarities with our isolates, including two isolates from Ghana and Taiwan (GHA8 and VGC1) and one from Japan (SC955).

4. Discussion

The Staphylococcus aureus is a common bacterial pathogen that can cause severe infectious diseases in livestock animals and humans. S. aureus may cause health problems, such as skin and soft tissue infection, septic arthritis, osteomyelitis, pneumonia, and endocarditis [30]. In this study, we found that 35% of the food products and human pus samples carried the coagulase-positive S. aureus (CPS). The CPS isolates showed resistance to amoxicillin (91.4%), penicillin (97.1%), cefoxitin (85.7%), cephradine (82.9%), tetracycline (57.2%), and erythromycin (52.8%).
The prevalence of S. aureus in minced and beef luncheon was 48% and 84%, respectively. Tang and colleagues [31] reported similar findings, detecting S. aureus in 68% of Denmark’s retail meat samples. In addition, a similar study identified S. aureus in 51%, 43.35%, and 12.2% of raw meat, quick-frozen meat, and ready to eat meat samples, respectively [32]. A low prevalence of S. aureus (16.6%) in meat samples from 60 butcher shops was reported in Egypt [33]. Furthermore, S. aureus prevalence of 27.9% was reported in retail meat samples in the USA [34].
In the present study, S. aureus was predominant in Karish cheese (76%). This result was consistent with other Egyptian studies that reported S. aureus prevalence rates of 93% and 90% from Karish cheese [35,36]. However, a lower S. aureus prevalence of 30% and 24% in Karish cheese in Egypt was also recorded [1,37]. The prevalence rate of S. aureus in human pus samples was 38% in the present study. This is in line with a previous study [38], which isolated S. aureus in 11.3% of patients from sputum specimens. The wide range in the reported prevalence of S. aureus in meat, milk products, and human samples among local and global studies may be attributed to differences in the hygienic measures implemented during food processing and manufacturing in the region of the study. Additionally, the handling procedure and hygienic condition surrounding the product manufacture can affect the degree and percentage of microbial contamination; for example, in Karish cheese, which is a type of handmade cheese (made of raw milk or laban rayeb with salt, produced without heat treatment) in popular use and is manufactured in most areas in Egypt. Furthermore, minced meat and beef luncheon are also manufactured from frozen or raw meat and the processing involves no or low heat treatment.
Based on the biochemical and enzymatic activity of the obtained coagulase-positive Staphylococci (CPS) isolates, hemolytic and DNase activities were detected in all sample types. Lecithinase activity was detected in minced meat, beef luncheon, and human samples; however, no Karish cheese isolates exhibited lecithinase activity. Similarly, Ali and colleagues [39] recorded DNase activity in 86.9% of the CPS. Furthermore, DNase activity and coagulase activity was observed among 60% and 87.5% of the CPS, respectively, as reported previously [37,40].
The molecular identification of the coa gene in S. aureus strains was performed through amplification of the coa gene at (600–1000) bp. The use of the coa gene to detect S. aureus strains from milk origin was previously reported in two studies [37,41] which amplified the coa gene from mastitic milk and dairy products at 630 and 750 bp. Moreover, Javid and colleagues [42] detected the coa gene at 514 bp, 595 bp, 757 bp, and 802 bp.
In recent years, the widespread use of antibiotics to treat bacterial infections has led to the emergence of multidrug-resistant (MDR) bacteria, which constitute a great hazard to public health. For example, S. aureus could adapt to adverse environmental conditions, leading to the emergence of many strains resistant to different antibiotic classes [43]. In the current study, the phenotypic testing of 70 S. aureus isolates from meat and milk products, and humans revealed a high prevalence of resistance against penicillin (97.1%), amoxicillin (91.4%), cefoxitin (85.7%), cephradine (82.9%), tetracycline (57.2%), and erythromycin (52.8%). Similar findings were obtained by Mohammed and Hafez [33] in Egypt, who detected high resistance against chloramphenicol and ampicillin (100%), streptomycin (66.6%), gentamicin and trimethoprim-sulphamethoxazole (33.3%), and erythromycin (16.6%). In a recent study in Egypt, S. aureus demonstrated extreme resistance to cefoxitin, penicillin, cephradine, tetracycline, trimethoprim-sulphamethoxazole, and norfloxacin in samples from raw milk and ice cream [4]. In China, Wu and colleagues [32] also recorded high resistance of S. aureus isolated from retail meat and meat products against penicillin (84.6%), ampicillin (85.4%), erythromycin (52.7%), tetracycline (49.3%), kanamycin (45.3%), telithromycin (30.1%), clindamycin (29.6%), streptomycin (21.1%), norfloxacin (20.4%), gentamicin (19.4%), fusidic acid (18.4%), and ciprofloxacin (16.9%). Most of the published studies supported our findings that S. aureus has become a notorious pathogen due to its high resistance against many antimicrobial agents. Therefore, more studies are required to overcome this problem in veterinary or human practices.
Methicillin-resistant S. aureus (MRSA) is a pathogen causing human and livestock animal infections. MRSA contaminates many types of food and dairy products, which cause foodborne poisoning [44] and the detection of MRSA in animal-derived foods is necessary for the monitoring and improvement of hygiene measures in food practices to diminish the potential for bacterial contamination [44]. The results in Figure 1 demonstrated a high prevalence and emergence of MRSA strains from meat, milk products, and human samples, constituting a major public health hazard. These are consistent with several other studies [45,46]. Interestingly, the existence of MRSA in the environment can generally allow the dissemination and spread of MRSA in animal products [47].
The mecA gene is known to encode penicillin-binding protein 2a (PBP2a), which is responsible for resistance against beta-lactam [16]. Modern molecular techniques, such as PCR, are used for mecA gene detection and is recognized as a specific method for the detection of MRSA strains among staphylococcal species [48]. In this study, the mecA gene was successfully amplified in 11 out of 19 tested isolates (57.89%) at 583 bp. A comparative study in Nigeria reported that the mecA gene was prevalent in 38% of S. aureus isolates from human clinical specimens [49]. In Egypt, 8 out of 19 (42.1%) food origin S. aureus isolates exhibited phenotypic resistance to oxacillin and carried the mecA gene [50]. A similar finding was reported by Awad and colleagues [51], who showed that 50% of S. aureus isolates of mastitis origin harbored the specific mecA gene and expressed phenotypic resistance to multiple antimicrobials groups. In addition, the existence of the mecA-positive MRSA in bovine milk strains was described earlier in the USA [52] as well as in Denmark [31].
The sequences analysis of the mecA gene originated from three different sources in this study (minced meat, Karish cheese, and human pus) revealed high genetic similarities (99.79 to 100%) with many isolates from different nations and localities both at nucleotides and protein level. This result was consistent with Malik and colleagues [53], who reported that identical homologs of the mecA gene of MRSA strains originated from human, cat, and dog, which is attributed to the close contact between humans and these animals. Moreover, these resistance genes are disseminated between animals and humans due to the normal inhabitance of Staphylococci in the skin and respiratory mucosa, which could easily be dispersed by aerosols from sneezing and coughing, skin-to-skin contact, and saliva [53]. In a Brazilian study, phylogenetic analysis of the mecA gene nucleotide sequences from 210 skin and ears rodent samples and 60 equine nasal samples separated them into two distinct groups in the phylogenic tree, one of which belongs to the bovine clones and the other related to the equine and human strains [54]. Interestingly, the authors declared that the mecA from equine S. sciuri and S. lentus was highly homologous to the mecA from human S. aureus, whereas a clear diversity from bovine S. sciuri was noticed, suggesting that the evolution of a resistance gene is more likely related to the host environment rather than the bacterial species. However, the presence of another mobile genetic element called staphylococcal chromosome cassette mec (SCCmec) in MRSA strains of human origin suggests that the mecA gene could be a shared gene among different staphylococcal species [55]. Regarding the dissemination of the mecA gene in both human and food samples, Velasco and colleagues [56] demonstrated the similarity between 108 clinical MRSA isolates and 133 S. aureus isolates from animal origin and clarified the high genetic similarity between the human and meat clones. The genetic similarity between human and meat strains suggests contamination of raw meat during handling and the potential risk of S. aureus spread through the food chain. High genetic similarity among S. aureus strains from different sources such as meat-producing animals and retail meat could be attributed to meat contamination during slaughtering [57]. However, Sung and colleagues [58] showed that clones of S. aureus among animals are closely related to human clones. This could be due to the adaptive behavior mechanism of S. aureus isolates [59]. In Iraq, Neamah and colleagues [60] found a high genetic linkage between S. aureus from humans and cattle and a major genetic link between livestock animals and human isolates.

5. Conclusions

The S. aureus was prevalent among meat, milk products, and human samples. Our results highlighted the predominance of MRSA strains with resistance to multiple antimicrobial agents in animal food products. In addition, the genetic analysis of the mecA gene revealed high nucleotide and amino acid similarities between S. aureus strains from human samples and food products. Additionally, the study detected the presence of the mecA gene in S. aureus isolates from humans and animal food products (minced meat and Karish cheese) in the study area in Egypt and its possible threat to public health. Therefore, frequent monitoring of S. aureus to identify patterns of antimicrobial susceptibility is needed to elucidate MRSA transmission routes through the animal food chain. In addition, it is necessary to follow up the hygienic measures to prevent or minimize the contamination of food products, particularly that of handmade products in the study area, which constitute a high risk on public health.

Supplementary Materials

The following are available online at https://0-www-mdpi-com.brum.beds.ac.uk/2079-6382/10/1/75/s1, Table S1: Multi-drug resistant S. aureus strains from food and human samples. Figure S1. Amplification of the coa gene of Staphylococcus aureus at 600–1000 bp: Lane M: 100 bp DNA ladder. Pos.: Positive control. Neg.: Negative control. Samples 1 and 2 from minced meat, 3 and 4 from beef luncheon, 5 and 6 from Karish cheese, and 7, 8, and 9 from human were recorded as positive samples for the coa gene. Supplementary Figure S2. Amplification of the 583 bp fragment of the mecA gene of Staphylococcus aureus; Lane M: 100 bp DNA ladder. Pos.: Positive control. Neg.: Negative control; Lane: Samples 3 and 5 from minced meat; 7 and 8 from beef luncheon; 9 and 12 from Karish cheese; 13, 14, 15, 18, and 19 from human samples were recorded as positive samples for the mecA gene.

Author Contributions

Conceptualization, E.E.A., W.S.M., and S.Y.A.; methodology, E.E.A., W.S.M., S.Y.A., H.S.H., and R.R.S.; software, M.M.S., M.N, and A.A.; validation, E.E.A., H.S.H., and A.H.; formal analysis, M.N., A.H., and A.A.; investigation, E.E.A., W.S.M., S.Y.A., H.S.H., and R.R.S.; resources, M.M.S. and A.H.; data curation, E.E.A. and M.N.; writing—original draft preparation, E.E.A., W.S.M., M.M.S., A.H., and A.A.; writing—review and editing, S.Y.A., H.S.H., M.N., R.R.S., G.E.B., and A.A.; visualization, H.S.H., M.N., and G.E.B. ; supervision, E.E.A. and W.S.M.; project administration, E.E.A.; funding acquisition, M.M.S. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Taif University Researchers Supporting Project (TURSP-2020/09), Taif University, Taif, Saudi Arabia.

Institutional Review Board Statement

This study was performed according to the rules and regulations of the Egyptian Ministry of Health and Faculty of Veterinary of Medicine, University of Sadat City, Egypt (Approval no. VUSC00006).

Informed Consent Statement

Informed consent was obtained from all subjects involved in the study.

Data Availability Statement

All authors agree that the data presented in this study are openly available through mdpi publisher platform or others without any restriction.

Acknowledgments

The authors extend their appreciation and sincere thanks to the Chief Director and the staff members at the Al-Qaluobyia Hospital for supplying clinical information and samples collection. We appreciate and thank Taif University for the financial support for Taif University Researchers Supporting Project (TURSP-2020/09), Taif University, Taif, Saudi Arabia.

Conflicts of Interest

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

References

  1. Mousa, W.S.; Abdeen, E.; Hussein, H.; Hadad, G. Prevalence and multiplex PCR for enterotoxin genes of Staphylococcus aureus isolates from subclinical mastitis and Kareish cheese. J. Anc. Dis. Prev. Remedies 2017, 5. [Google Scholar] [CrossRef]
  2. Velázquez-Ordoñez, V.; Valladares-Carranza, B.; Tenorio-Borroto, E.; Talavera-Rojas, M.; Antonio Varela-Guerrero, J.; Acosta-Dibarrat, J.; Puigvert, F.; Grille, L.; González Revello, Á.; Pareja, L. Microbial contamination in milk quality and health risk of the consumers of raw milk and dairy products. In Nutrition in xHealth and Disease—Our Challenges Now and Forthcoming Time; Mózsik, G., Figler, M., Eds.; IntechOpen: London, UK, 2019; ISBN1 978-1-78984-007-0. ISBN2 978-1-78984-008-7. [Google Scholar]
  3. Paterson, G.K.; Morgan, F.J.E.; Harrison, E.M.; Peacock, S.J.; Parkhill, J.; Zadoks, R.N.; Holmes, M.A. Prevalence and properties of mecc methicillin-resistant Staphylococcus aureus (mrsa) in bovine bulk tank milk in great britain. J. Antimicrob. Chemother. 2014, 69, 598–602. [Google Scholar] [CrossRef] [PubMed]
  4. Abdeen, E.E.; Mousa, W.S.; Abdel Salam, S.Y.; Al-Maary, K.S.; Mubarak, A.S.; Moussa, I.M.; Hemeg, H.A.; Almuzaini, A.M.; Alajaji, A.I.; Alsubki, R.A.; et al. Antibiogram and phylogenetic diversity of enterotoxigenic Staphylococcus aureus strains from milk products and public health implications. Saudi J. Biol. Sci. 2020, 27, 1968–1974. [Google Scholar] [CrossRef] [PubMed]
  5. Akabanda, F.; Hlortsi, E.H.; Owusu-Kwarteng, J. Food safety knowledge, attitudes and practices of institutional food-handlers in Ghana. BMC Public Health 2017, 17. [Google Scholar] [CrossRef]
  6. Kalorey, D.R.; Shanmugam, Y.; Kurkure, N.V.; Chousalkar, K.K.; Barbuddhe, S.B. PCR-based detection of genes encoding virulence determinants in Staphylococcus aureus from bovine subclinical mastitis cases. J. Vet. Sci. 2007, 8, 151. [Google Scholar] [CrossRef]
  7. Zecconi, A.; Scali, F. Staphylococcus aureus virulence factors in evasion from innate immune defenses in human and animal diseases. Immunol. Lett. 2013, 150, 12–22. [Google Scholar] [CrossRef]
  8. Dallal, M.M.S.; Khoramizadeh, M.R.; Amiri, S.A.; Yaraghi, A.A.S.; Fard, R.M.N. Coagulase gene polymorphism of Staphylococcus aureus isolates: A study on dairy food products and other foods in Tehran, Iran. Food Sci. Hum. Wellness 2016, 5, 186–190. [Google Scholar] [CrossRef]
  9. van Cleef, B.A.G.L.; Monnet, D.L.; Voss, A.; Krziwanek, K.; Allerberger, F.; Struelens, M.; Zemlickova, H.; Skov, R.L.; Vuopio-Varkila, J.; Cuny, C.; et al. Livestock-associated methicillin-resistant staphylococcus aureu s in humans, Europe. Emerg. Infect. Dis. 2011, 17, 502–505. [Google Scholar] [CrossRef]
  10. Thapaliya, D.; Forshey, B.M.; Kadariya, J.; Quick, M.K.; Farina, S.; O’ Brien, A.; Nair, R.; Nworie, A.; Hanson, B.; Kates, A.; et al. Prevalence and molecular characterization of Staphylococcus aureus in commercially available meat over a one-year period in Iowa, USA. Food Microbiol. 2017, 65, 122–129. [Google Scholar] [CrossRef]
  11. Abdel-Maksoud, M.; El-Shokry, M.; Ismail, G.; Hafez, S.; El-Kholy, A.; Attia, E.; Talaat, M. Methicillin-resistant Staphylococcus aureus recovered from healthcare- and community-associated infections in Egypt. Int. J. Bacteriol. 2016, 2016, 1–5. [Google Scholar] [CrossRef]
  12. Bhargava, K.; Wang, X.; Donabedian, S.; Zervos, M.; da Rocha, L.; Zhang, Y. Methicillin-resistant Staphylococcus aureus in retail meat, Detroit, Michigan, USA. Emerg. Infect. Dis. 2011, 17, 1135–1137. [Google Scholar] [CrossRef] [PubMed]
  13. Jans, C.; Merz, A.; Johler, S.; Younan, M.; Tanner, S.A.; Kaindi, D.W.M.; Wangoh, J.; Bonfoh, B.; Meile, L.; Tasara, T. East and west African milk products are reservoirs for human and livestock-associated Staphylococcus aureus. Food Microbiol. 2017, 65, 64–73. [Google Scholar] [CrossRef] [PubMed]
  14. Abubakar, U.; Sulaiman, S.A.S. Prevalence, trend and antimicrobial susceptibility of Methicillin Resistant Staphylococcus aureus in Nigeria: A systematic review. J. Infect. Public Health 2018, 11, 763–770. [Google Scholar] [CrossRef]
  15. Okorie-Kanu, O.J.; Anyanwu, M.U.; Ezenduka, E.V.; Mgbeahuruike, A.C.; Thapaliya, D.; Gerbig, G.; Ugwuijem, E.E.; Okorie-Kanu, C.O.; Agbowo, P.; Olorunleke, S.; et al. Molecular epidemiology, genetic diversity and antimicrobial resistance of Staphylococcus aureus isolated from chicken and pig carcasses, and carcass handlers. PLoS ONE 2020, 15. [Google Scholar] [CrossRef]
  16. Arsic, B.; Zhu, Y.; Heinrichs, D.E.; McGavin, M.J. Induction of the Staphylococcal proteolytic cascade by antimicrobial fatty acids in community acquired methicillin resistant Staphylococcus aureus. PLoS ONE 2012, 7. [Google Scholar] [CrossRef]
  17. Maes, N.; Magdalena, J.; Rottiers, S.; De Gheldre, Y.; Struelens, M.J. Evaluation of a triplex PCR assay to discriminate Staphylococcus aureus from coagulase-negative staphylococci and determine methicillin resistance from blood cultures. J. Clin. Microbiol. 2002, 40, 1514–1517. [Google Scholar] [CrossRef]
  18. García-Álvarez, L.; Holden, M.T.G.; Lindsay, H.; Webb, C.R.; Brown, D.F.J.; Curran, M.D.; Walpole, E.; Brooks, K.; Pickard, D.J.; Teale, C.; et al. Meticillin-resistant Staphylococcus aureus with a novel mecA homologue in human and bovine populations in the UK and Denmark: A descriptive study. Lancet Infect. Dis. 2011, 11, 595–603. [Google Scholar] [CrossRef]
  19. CAPMAS. The Result of Fifth Economic Census 2017–2018 the Total Egypt to Economic Activity and Governorates; CAPMAS: Cairo, Egypt, 2018. [Google Scholar]
  20. APHA. Standard Methods for the Examination of Dairy Products, 15th ed.; Americana Public Health Association: Washington, DC, USA, 1992. [Google Scholar]
  21. Rice, M.; Turner-Henson, A.; Park, N.-J.; Azuero, A.; Amiri, A.; Feeley, C.A.; Johnson, A.; Lam, T.; Huntington-Moskos, L.; Rodriguez, J.; et al. Child and maternal factors that influence child blood pressure in preschool children: An exploratory study. Appl. Nurs. Res. 2016, 31, 117–120. [Google Scholar] [CrossRef]
  22. Murray, D.P.R.; Baron, E.J.; Jorgensen, J.H. Manual of Clinical Microbiology, 8th ed.; American Society for Microbiology: Washington, DC, USA, 2003. [Google Scholar]
  23. CLSI. Performance Standards for Antimicrobial Susceptibility Testing; CLSI: Tucson, AZ, USA, 2017; ISBN 978-1-56238-804-1. [Google Scholar]
  24. Azimian, A.; Havaei, S.A.; Fazeli, H.; Naderi, M.; Ghazvini, K.; Samiee, S.M.; Soleimani, M.; Peerayeh, S.N. Genetic characterization of a vancomycin-resistant Staphylococcus aureus isolate from the respiratory tract of a patient in a University Hospital in Northeastern Iran. J. Clin. Microbiol. 2012, 50, 3581–3585. [Google Scholar] [CrossRef]
  25. Aarestrup, F.M.; Dangler, C.A.; Sordillo, L.M. Prevalence of coagulase gene polymorphism in Staphylococcus aureus Isolates causing bovine mastitis. Can. J. Vet. Res. 1995, 59, 124–128. [Google Scholar]
  26. Hall, T.A. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 1999, 41, 95–98. [Google Scholar]
  27. Thompson, J.D.; Higgins, D.G.; Gibson, T.J. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994, 22, 4673–4680. [Google Scholar] [CrossRef]
  28. Higgins, D.G.; Sharp, P.M. Fast and sensitive multiple sequence alignments on a microcomputer. Bioinformatics 1989, 5, 151–153. [Google Scholar] [CrossRef] [PubMed]
  29. Kumar, S. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief. Bioinform. 2004, 5, 150–163. [Google Scholar] [CrossRef] [PubMed]
  30. McDougal, L.K.; Steward, C.D.; Killgore, G.E.; Chaitram, J.M.; McAllister, S.K.; Tenover, F.C. Pulsed-field gel electrophoresis typing of oxacillin-resistant Staphylococcus aureus isolates from the United States: Establishing a national database. J. Clin. Microbiol. 2003, 41, 5113–5120. [Google Scholar] [CrossRef] [PubMed]
  31. Tang, Y.; Larsen, J.; Kjeldgaard, J.; Andersen, P.S.; Skov, R.; Ingmer, H. Methicillin-resistant and -susceptible Staphylococcus aureus from retail meat in Denmark. Int. J. Food Microbiol. 2017, 249, 72–76. [Google Scholar] [CrossRef] [PubMed]
  32. Wu, S.; Huang, J.; Wu, Q.; Zhang, J.; Zhang, F.; Yang, X.; Wu, H.; Zeng, H.; Chen, M.; Ding, Y.; et al. Staphylococcus aureusisolated from retail meat and meat products in China: Incidence, antibiotic resistance and genetic diversity. Front. Microbiol. 2018, 9. [Google Scholar] [CrossRef]
  33. Mohammed, G.M.O.; Hafez, T.A. Detection of mecA gene in methicillin-resistance Staphylococcus aureus (MRSA) strains isolated from beef meat using polymerase chain reaction. Assiut Vet. Med. J. 2016, 62, 64–70. [Google Scholar]
  34. Ge, B.; Mukherjee, S.; Hsu, C.-H.; Davis, J.A.; Tran, T.T.T.; Yang, Q.; Abbott, J.W.; Ayers, S.L.; Young, S.R.; Crarey, E.T.; et al. MRSA and multidrug-resistant Staphylococcus aureus in U.S. retail meats, 2010–2011. Food Microbiol. 2017, 62, 289–297. [Google Scholar] [CrossRef]
  35. El-Malt, L.M. Occurrence of enterotoxigenic staphylococcus aureus in some cheese varieties in Aswan city —Upper Egypt. Assiut Vet. Med. J. 2013, 59, 62–70. [Google Scholar]
  36. Salem, H.; El-Attar, L.; Omran, E. Microbiological Assessment of some parameters of Kariesh cheese sold by supermarkets and street vendors in Alexandria, Egypt. J. High Inst. Public Health 2016, 46, 77–85. [Google Scholar] [CrossRef]
  37. Elmaghraby, M.; Abdeen, E.; AbouElros, N.; Bagory, A. Molecular identification of virulence genes of Staphylococcus aureus isolated from milk products. Alex. J. Vet. Sci. 2018, 59, 57. [Google Scholar] [CrossRef]
  38. Metersky, M.L.; Aksamit, T.R.; Barker, A.; Choate, R.; Daley, C.L.; Daniels, L.A.; DiMango, A.; Eden, E.; Griffith, D.; Johnson, M.; et al. The prevalence and significance of Staphylococcus aureus in patients with non-cystic fibrosis bronchiectasis. Ann. Am. Thorac. Soc. 2018, 15, 365–370. [Google Scholar] [CrossRef] [PubMed]
  39. Ali, A.; Mahmoud, H.; Sameh, A.; Zaky, S.; Elsayed, A. Antibiotic resistance profile of coagulase positive Staphylococcal infection in dairy buffaloe. Worlds Vet. J. 2015, 6, 46. [Google Scholar] [CrossRef]
  40. Abdel All, A.; Bashandy, M.M.; Yasin, M.H.; Ibrahim, A.K. Assessment of conventional and molecularfeatures of Staphylococcus aureus isolated from bovine milk samples and contact dairy workers. Glob. Vet. 2010, 4, 168–175. [Google Scholar]
  41. Abdeen E., E.; Walid, M.; Hussien, H.; Roshdy, S. PCR for detection of virulence and antibiotic resistance genes of coagulase positive staphylococcus aureus from clinical mastitis in Egypt. Int. J. Basic Appl. Sci. 2015, 4, 315. [Google Scholar] [CrossRef]
  42. Javid, F.; Taku, A.; Bhat, M.A.; Badroo, G.A.; Mudasir, M.; Sofi, T.A. Molecular typing of Staphylococcus aureus based on coagulase gene. Vet. World 2018, 11, 423–430. [Google Scholar] [CrossRef]
  43. McCallum, N.; Berger-Bächi, B.; Senn, M.M. Regulation of antibiotic resistance in Staphylococcus aureus. Int. J. Med. Microbiol. 2010, 300, 118–129. [Google Scholar] [CrossRef]
  44. Basanisi, M.G.; La Bella, G.; Nobili, G.; Franconieri, I.; La Salandra, G. Genotyping of methicillin-resistant Staphylococcus aureus (MRSA) isolated from milk and dairy products in South Italy. Food Microbiol. 2017, 62, 141–146. [Google Scholar] [CrossRef]
  45. Lin, Q.; Sun, H.; Yao, K.; Cai, J.; Ren, Y.; Chi, Y. The prevalence, antibiotic resistance and biofilm formation of Staphylococcus aureus in bulk ready- to-eat foods. Biomolecules 2019, 9, 524. [Google Scholar] [CrossRef]
  46. Papadopoulos, P.; Papadopoulos, T.; Angelidis, A.S.; Kotzamanidis, C.; Zdragas, A.; Papa, A.; Filioussis, G.; Sergelidis, D. Prevalence, antimicrobial susceptibility and characterization of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus isolated from dairy industries in north-central and north-eastern Greece. Int. J. Food Microbiol. 2019, 291, 35–41. [Google Scholar] [CrossRef] [PubMed]
  47. Lim, S.-K.; Nam, H.-M.; Jang, G.-C.; Lee, H.-S.; Jung, S.-C.; Kim, T.-S. Transmission and persistence of methicillin-resistant Staphylococcus aureus in milk, environment, and workers in dairy cattle farms. Foodborne Pathog. Dis. 2013, 10, 731–736. [Google Scholar] [CrossRef] [PubMed]
  48. Al-Abbas, M.J.A. Antimicrobial susceptibility of Enterococcus faecalis and a novel Planomicrobium isolate of bacterimia. Int. J. Med. Med. Sci. 2012, 4. [Google Scholar] [CrossRef]
  49. Ibadin, E.E.; Enabulele, I.O.; Muinah, F. Prevalence of mecA gene among staphylococci from clinical samples of a tertiary hospital in Benin City, Nigeria. Afr. Health Sci. 2018, 17, 1000. [Google Scholar] [CrossRef] [PubMed]
  50. El Seedy, F.R.; Salam, H.S.H.; Aa, S.; Khairy, E.A.; Omara, S.T. Phenotypic and genotypic characterization of methicillin resistant Staphylococcus aureus. J. Vet. Med. Res. 2017, 24, 186–193. [Google Scholar] [CrossRef]
  51. Awad, A.; Ramadan, H.; Nasr, S.; Ateya, A.; Atwa, S. Genetic characterization, antimicrobial resistance patterns and virulence determinants of Staphylococcus aureus isolated form bovine mastitis. Pakistan J. Biol. Sci. 2017, 20, 298–305. [Google Scholar] [CrossRef]
  52. Haran, K.P.; Godden, S.M.; Boxrud, D.; Jawahir, S.; Bender, J.B.; Sreevatsan, S. Prevalence and characterization of Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus, isolated from bulk tank milk from Minnesota dairy farms. J. Clin. Microbiol. 2012, 50, 688–695. [Google Scholar] [CrossRef]
  53. Malik, S.; Peng, H.; Barton, M.D. Partial nucleotide sequencing of the mecA genes of Staphylococcus aureus isolates from cats and dogs. J. Clin. Microbiol. 2006, 44, 413–416. [Google Scholar] [CrossRef]
  54. Silva, N.C.C.; Guimarães, F.F.; Manzi, M.P.; Júnior, A.F.; Gómez-Sanz, E.; Gómez, P.; Langoni, H.; Rall, V.L.M.; Torres, C. Methicillin-resistant Staphylococcus aureus of lineage ST398 as cause of mastitis in cows. Lett. Appl. Microbiol. 2014, 59, 665–669. [Google Scholar] [CrossRef]
  55. Katayama, Y.; Ito, T.; Hiramatsu, K. A new class of genetic element, staphylococcus cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus. Antimicrob. Agents Chemother. 2000, 44, 1549–1555. [Google Scholar] [CrossRef]
  56. Velasco, V.; Buyukcangaz, E.; Sherwood, J.S.; Stepan, R.M.; Koslofsky, R.J.; Logue, C.M. Characterization of Staphylococcus aureus from humans and a comparison with isolates of animal origin in North Dakota, United States. PLoS ONE 2015, 10, e0140497. [Google Scholar] [CrossRef] [PubMed]
  57. Buyukcangaz, E.; Velasco, V.; Sherwood, J.S.; Stepan, R.M.; Koslofsky, R.J.; Logue, C.M. Molecular typing of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) isolated from animals and retail meat in North Dakota, United States. Foodborne Pathog. Dis. 2013, 10, 608–617. [Google Scholar] [CrossRef] [PubMed]
  58. Sung, J.M.L.; Lloyd, D.H.; Lindsay, J.A. Staphylococcus aureus host specificity: Comparative genomics of human versus animal isolates by multi-strain microarray. Microbiology 2008, 154, 1949–1959. [Google Scholar] [CrossRef] [PubMed]
  59. McCarthy, A.J.; Lindsay, J.A.; Loeffler, A. Are all meticillin-resistant Staphylococcus aureus (MRSA) equal in all hosts? Epidemiological and genetic comparison between animal and human MRSA: Evolution of meticillin-resistant Staphylococcus aureus. Vet. Dermatol. 2012, 23, 267-e54. [Google Scholar] [CrossRef]
  60. Neamah, A.J.; Ayyez, H.N.; Klaif, S.F.; Khudhair, Y.I.; Hussain, M.H. Molecular and phylogenetic study of Staphylococcus aureus isolated from human and cattle of Al-Qadisiyah Governorate, Iraq. Vet. World 2019, 12, 1378–1382. [Google Scholar] [CrossRef]
Figure 1. An UpSetR plot is showing multidrug-resistant profile among the 70 Staphylococcus aureus strains detected in animal food products and human pus samples. Note: penicillin (P), amoxicillin (AX), cefoxitin (FOX), vancomycin (VA), gentamicin (CN), erythromycin (E), tetracycline (TE), ciprofloxacin (CIP), norfloxacin (NOR), trimethoprim-sulfamethoxazole (SXT), chloramphenicol (C), cephradine (CE).
Figure 1. An UpSetR plot is showing multidrug-resistant profile among the 70 Staphylococcus aureus strains detected in animal food products and human pus samples. Note: penicillin (P), amoxicillin (AX), cefoxitin (FOX), vancomycin (VA), gentamicin (CN), erythromycin (E), tetracycline (TE), ciprofloxacin (CIP), norfloxacin (NOR), trimethoprim-sulfamethoxazole (SXT), chloramphenicol (C), cephradine (CE).
Antibiotics 10 00075 g001
Figure 2. Dendrogram for nucleotide similarity between Staphylococcus aureus isolates from minced meat, Karish cheese, and human samples. Sequences (highlighted in blue color) from human samples (MT185692), minced meat (MT 211620), and Karish cheese (MT 211621) were generated in this study.
Figure 2. Dendrogram for nucleotide similarity between Staphylococcus aureus isolates from minced meat, Karish cheese, and human samples. Sequences (highlighted in blue color) from human samples (MT185692), minced meat (MT 211620), and Karish cheese (MT 211621) were generated in this study.
Antibiotics 10 00075 g002
Figure 3. Dendrogram for protein similarity between Staphylococcus aureus isolates from minced meat, Karish cheese, and human samples. Sequences (highlighted in blue color) from human samples (MT185692), minced meat (MT 211620), and Karish cheese (MT 211621) were generated in this study.
Figure 3. Dendrogram for protein similarity between Staphylococcus aureus isolates from minced meat, Karish cheese, and human samples. Sequences (highlighted in blue color) from human samples (MT185692), minced meat (MT 211620), and Karish cheese (MT 211621) were generated in this study.
Antibiotics 10 00075 g003
Table 2. Prevalence of coagulase-positive Staphylococci (CPS) isolated from food and human samples.
Table 2. Prevalence of coagulase-positive Staphylococci (CPS) isolated from food and human samples.
Human Swabs
(n = 50)
Karish Cheese
(n = 50)
Beef Luncheon
(n = 50)
Minced Meat
(n = 50)
CPSCPSCPSCPS
%No.%No.%No.%No.
3819763884424824
% estimated according to the number of each sample (n = 50).
Table 3. Antimicrobial resistance of the CPS isolates.
Table 3. Antimicrobial resistance of the CPS isolates.
AntibioticsAntimicrobial ClassesResistantIntermediateSensitive
No.%No.%No.%
Penicillin (P) 1 IUβ-lactams6897.10022.9
Amoxicillin (AX) 25 µgβ-lactams6491.40068.6
Cefoxitin (FOX) 30 µgβ-lactams6085.7001014.3
Vancomycin (VA) 30 µgGlycopeptides1014.311.45984.3
Gentamicin (CN) 10 µgAminoglycosides3042.8912.93144.3
Erythromycin (E) 15 µgMacrolides3752.83042.934.3
Tetracycline (TE) 30 µgTetracyclines4057.2811.42231.4
Ciprofloxacin (CIP) 5 µgFluoroquinolones2332.91724.33042.8
Norfloxacin (NOR) 10 µgFluoroquinolones1724.314203955.7
Trimethoprim-Sulfamethoxazole (SXT) 1.25/23.75 µgSulfonamides2941.468.63550
Chloramphenicol (C) 30 µgChloramphenicol1825.734.34970
Cephradine (CE) 30 µgβ-lactams5882.9001217.1
% estimated according to the number of tested isolates for antibiotic sensitivity test (n = 70).
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Back to TopTop