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Phylogenetic Tracking of Ica-locus in MRSA from Different Sources at Ismailia City, Egypt

AAVS_10_10_2124-2132

Research Article

Phylogenetic Tracking of Ica-locus in MRSA from Different Sources at Ismailia City, Egypt

Ali Wahdan1*, Mahmoud Ezzat1, Fatma Youssef2, Manal Munier2, Elsayyad M. Ahmed3, Marwa Abo Hashem1

1Department of Bacteriology, Immunology and Mycology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt; 2Department of Clinical Pathology, Animal Health Research Institute, Ismailia, Egypt; 3Department of Virology, Animal Health Research Institute (AHRI), Dokki, Giza, Egypt.

Abstract | Tracking of MRSA strains which encoded intracellular adhesion (ica) locus is a major threat for livestock and public health. As it indicates that the isolated strains have ability to biofilm formation leading to zoonotic risk and nosocomial infections. So, this study focused on tracking the ica locus in different hosts and other virulence factors with genetic analysis of recovered isolates. A total of 350 random samples were collected from animals and hospital acquired infected patients. The recovered isolates were identified biochemically by vitek2 compact, confirmed by PCR and tested for sensitivity against varieties of antimicrobials. Three multi-drug resistant isolates carried ica from human abscess (A9), sputum of dog (A12), and cow mastitic milk (A13) were sequenced with other genome sequences retrieved from the GenBank. Ten samples (9.17%) were S. aureus from human, while 3 samples (1.24%) were S. aureus and 2 samples (0.83%) were coagulase negative staphylococcus (CONS) from animal sources. The multiple sequences alignments (MSA) and phylogenetic tracking proved the genetic similarity among A9, A12 and A13 from (human, animal, food of animal origin respectively) reached to 94 %. In conclusion, although the low isolation rate of confirmed MRSA in Ismailia governorate, the three selected MRSA strains invigorate the possibility of transmission among different sources of food, animals, and human, which causes nosocomial infections.

 

Keywords | Ica-MRSA, PCR, Virulence factors, Resistant factors, Phylogenomics.


Received | August 11, 2022; Accepted | September 01, 2022; Published | September 15, 2022

*Correspondence | Ali Wahdan, Department of Bacteriology, Immunology and Mycology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt; Email: dr_aly_w@yahoo.com

Citation | Wahdan A, Ezzat M, Youssef F, Munier M, Ahmed SM, Hashem MA (2022). Phylogenetic tracking of Ica-locus in MRSA from different sources at ismailia city, egypt.. Adv. Anim. Vet. Sci. 10(10): 2124-2132.

DOI | http://dx.doi.org/10.17582/journal.aavs/2022/10.10.2124.2132

ISSN (Online) | 2307-8316

 

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Copyright: 2022 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).



INTRODUCTION

Staphylococci are common bacterial pathogens that may infect both animals and humans, resulting in a high number of hospital and community-acquired diseases each year. Staphylococcal infections have a significant economic and clinical impact due to a range of factors, including increased antibiotic resistance and a lack of effective immunizations (Asante et al., 2020).

Staphylococcus aureus is a prevalent human and animal infection that causes a variety of clinical symptoms (Tong et al., 2015); (Becker et al., 2017). S. aureus and other Staphylococcal species are considered public health pathogens due to their ability to rapidly acquire multiple resistance genes (Lakhundi and Zhang, 2018). Thus, methicillin-resistant S. aureus (MRSA) and coagulase-negative Staphylococci are among the most common causes of healthcare-associated infections (MR-CoNS) (Becker et al., 2020).

Staphylococcus aureus causes major difficulties in animals, including suppurated illness, mastitis, arthritis, and urinary tract infection. In humans, S. aureus is a major cause of food poisoning, pneumonia, surgical wound infections, and nosocomial bacteremia. (John and Harvin, 2007). Meanwhile, CoNS, especially S. epidermidis and S. haemolyticus, have emerged as prevalent causative agents of nosocomial infections, particularly those linked with indwelling devices. Because they actively assist in horizontal resistance transmission, they offer a significant concern in the final stages of multidrug resistance (Becker et al., 2014).

Staphylococcus aureus generates a diverse set of extracellular proteins and polysaccharides, some of which are associated with virulence. Vaccines are not currently available, despite the fact that antibodies may neutralize Staphylococcal toxins and enzymes. Most antibiotics used to treat infections are resistant to some kinds of bacteria that infect hospitalized patients, with vancomycin being the only remaining medicine that has not acquired resistance (Foster, 1996) (Tong et al., 2015). Multidrug resistance strains can be discovered in persons, pets, foodstuff, other species, and the ecosystem (Sabat et al., 2013; Rossi et al., 2019).

Biofilm development allows germs to compete with human immunity while also protecting microbes from antimicrobial agents. This trait is associated with bacterial pathogenicity and several chronic bacterial diseases (Vancraeynest et al., 2004). Because they play a significant role in the production of polysaccharide intercellular adhesion, intercellular adhesion A and B (icaA and icaD) are responsible for Staphylococci biofilm growth (Fitzpatrick et al., 2005; Rohde et al., 2007).

Multiple of the genetic markers of resistance may have been exchanged among many staphylococcal species from varied environments and hosts, according to genomic studies (Xu et al., 2022). Due to local variations in control practices and specific characteristics of circulating clones, the overall geographic distribution of MRSA, this study focused on phylogenetic tracking the Ica locus in MRSA isolated from different sources with detection of some virulence and antibiotic resistance factors of the recovered isolates.

MATERIALS AND METHODS

Ethical Approval And Study Area

The investigation was carried out in accordance with Egyptian regulations on animal experimentation and veterinary medicine practice. The study is not an animal experiment, but rather the collection of samples in accordance with Egyptian Veterinary Practice. All human samples were collected with the permission of the Ismailia hospital. Veterinary samples were obtained from veterinary clinics, farms, and marketplaces throughout the governorates of Ismailia. Various areas in Egypt’s Ismailia governorate were studied between 2020 and 2022.

Sampling

Different samples were collected from different sources in sterile manner as follow; 25 g from raw meat, processed milk products and poultry meat, 5ml raw milk and 43 lesion swabs from farms and veterinary clinics. Twenty five blood samples, 30 urine samples and 54 swabs from lesions of infected patients at Ismailia hospital. All swabs were transported in 10 ml peptone water (Power and Johnson, 2009). Urine and other samples were transported in sterile container for bacteriological examination. All sampled were prepared and processed according to instructions of (Eaton et al., 1995) (Quinn et al., 2015).

Isolation And Vitek2 Characterization Of The Isolates

According to (Quinn et al., 2011) samples were grown in peptone water for 24 hours at 37 before being transferred to 5 % sheep blood agar and mannitol salt agar. After incubating all inoculation plates at 37°C for 24-48 hours, colonies were detected. Staphylococcus colonies were evaluated visually and biochemically by Vitek 2 compact utilizing Gram positive cards after the turbidity was adjusted to 0.53 to 0.63 according to (Rampacci et al., 2021).

Pcr Recognition Of Some Virulence And Resistance Genes

MRSA DNA was extracted from purified bacterial cells using the QIAamp DNA Mini Kit (Invitrogen, USA), and the methods were followed exactly (Ezzat et al., 2018). From the confirmed strains nuc,ica, spa, mecA, blaZ, Coa genes were detected, PCR was performed using specific sets of primers (Metabion, Germany), and the cycling conditions were performed as in (Table 1) according to (Ezzat et al., 2018). The final volume was 25 µl (12.5 µl Go Taq® Green Master Mix 2X, µl (20 pmol) for each forward and reverse primer, 5µl sample DNA, and PCR water up to 25 µl). The negative control was PCR water, and the positive control was graciously donated by A.H.R I, Dokki, Egypt). The proper bands were detected using 1% agarose gel electrophoresis at 100 Volt for 45 minutes (Applichem GmbH, Darmstadt, Germany) and photographed using a UV transilluminator. (Germany, Fermentas, Thermo Scientific), 100 bp ladder was utilized.

Alignment And Pylogenetic Comparison Of Ica Locus In Mrsa From Different Sources

Three isolates harbored to mecA and ica genes (human abscess (A9), sputum of dog (A12), and cow mastitic milk (A13) were aligned and sequenced. QIAquick PCR Product extraction kit was used to purify the PCR (Qiagen Inc. Valencia CA). Cycle sequencing kit Bigdye Terminator V3.1. Gene sequencing was conducted using an Applied

 

Table 1: Cycles conditions of PCR

Gene

Intial denaturation

denaturation Annealing Extension Cycles

Final extension

mecA

94˚C

5 min.

94˚C

30 sec.

50˚C

30 sec.

72˚C

30 sec.

35

72˚C

10 min.

coa

94˚C

5 min.

94˚C

30 sec.

55˚C

40 sec.

72˚C

45 sec.

35

72˚C

10 min.

spa

94˚C

5 min.

94˚C

30 sec.

55˚C

30 sec.

72˚C

30 sec.

35

72˚C

7 min.

blaZ

94˚C

5 min.

94˚C

30 sec.

50˚C

40 sec.

72˚C

50 sec.

35

72˚C

10 min.

icaA

94˚C

5 min.

94˚C

30 sec.

49˚C

1 min.

72˚C

1 min.

35

72˚C

10 min.

Nuc

94˚C

5 min.

94˚C

30 sec.

55˚C

40 sec.

72˚C

40 sec.

35

72˚C

10 min.

 

Table 2: Prevalence of negative and positive coagulase Staphylococcus isolated from different sources:

Species

Sources of the sample No. samples

Bacteriological finding

 

No. of coagulase positive Staph

%

No of coagulase negative Staph

%

Human

 

Lesion swabs (Wound, pus)

54 7 12.96 0 0

Blood cultures

25 2 8 0 0

Urine culture

30 1 3.33 0 0

Total

109 10 9.17 0 0
Animal raw meat 52 0 0

1 S. xylosus

1.92

raw milk

51 1 1.96

1 S. saprophyticus

1.96

milk product

50 0 0 0 0

Poultry

45 0 0 0 0

lesion swabs

43 2 4.6 0 0

Total

241

3

1.24

2

0.83

 

Table 3: Results of virulence and antibiotic resistant genes

 

Code

Source of samples

PCR results

nuc

ica

spa

mecA

blaZ

coa

Human origin

1

abscess

+

+ + + + +

2

abscess

+

+ + + _ _

3

blood

+

_ + + + _

4

blood

_

_ + + _ _

5

abscess

+

+ + + + +

6

abscess

+

_ + _ _ +

7

urine

+

_ + + + +

8

abscess

+

+ + + + +

9

abscess

+

+ + + + +

10

wound

+

+ + + + +
Animal origin

11

abscess from cat

+

+ + + + +

12

Sputum from dog

+

+ + + + +

13

Mastitic milk

+

+ + + + +

S.saprophyticus

S.xylosus

(CONS

14

Raw milk

-

-

-

-

-

-

15

Raw meat

-

-

-

-

-

-

Total

12

9

13

12

10

10

% Percent

80%

60%

86.6%

80%

66.6%

66.6%

 

Biosystems 3130 genetic analyzer (HITACHI, Japan) and a Perkin-Elmer cat-number (4336817) (Perkin-Elmer, Foster City, CA) as (Wahdan et al., 2020). Using the Basic local alignment tool for nucleotide (BLASTN), the sequences were collected from the NCBI, and high similar sequences for the ica gene were chosen and downloaded in fatsa format (Tamura et al., 2013). Multiple sequence alignments (MSA) were utilized in conjunction with the UGEN software and the MUSCLE method to investigate the evolutionary relationship between our isolates and strains in the GenBank database (Okonechnikov et al., 2012). The phylogenetic tree was constructed in IQ-TREE (Nguyen et al., 2015) using the maximum likelihood method, Model Finder to choose the best-fitting model, and values from 1000 bootstrap replicates. The tree was scaled to show the amount of substitutions per site, with branch length indicating how many substitutions there are per site. The evolutionary tree was shown, manipulated, and annotated using the Interactive Tree of Life (iTOL) (Letunic and Bork, 2021).

RESULTS

Regarding to phenotypic characterization, large, round, convex, often with beta hemolysis on blood agar and yellow colonies with yellow precipitation on mannitol salt agar were selected and confirmed biochemically. Among 350 specimens from humans, animals, food of animal origin examined in this study, 15 contained Staphylococcus with percentage 4.3% from different sources as shown in (Table 2). Ten S. aureus (9.17%) was recovered from 109 human samples. Seven isolates from wounds and pus swab samples, 2 isolates from blood and one isolate from urine samples. Three S. aureus (1.24%) and 2 staphylococcus (CONS) (0.83%) were recovered from 241 animal samples. All recovered colonies were confirmed and some virulence genes (nuc, ica, spa, mecA, blaZ, coa) were marked by PCR as shown in (Table 3). Twelve isolates (80%) were nuc gene positive, 9 isolates (60%) were ica gene positive, 13 isolates (86.6%) were spa gene positive, 12 isolates (80%) were mecA gene positive, 10 isolates (66.6%) were positive for both blaZ and coa genes. CONS were negative for all tested genes.

Three isolates harbored to mecA and ica genes (human abscess (A9), sputum of dog (A12), and cow mastitic milk (A13) were aligned and sequenced and taken accession numbers (On994655, On994656, and On994657) at GenBank. Multiple sequences alignments and phylogenetic analysis of (Figure 1), proved that the sequence identity was 94% between 3 selected S. aureus. Our isolates are

 

closely related to the isolates from different geographical locations, 96% identity with isolates from USA, India, Germany, Thiland, China, UK, Irland and 95% for 1 isolate from Kazakhstan, all clustered together. The phylogenetic tree of ica clustered the Egyptian three isolates with the other 33 S. aureus isolates. It proved that all have the same ica except for two nucleotides difference from C to G at position 144 in the isolates of Germany (CP022682.1 Homo Sapines) and Brazail (CP065869.1 Homo Sapines) other single nucleotide polymorphism (SNP) from C to T in isolates from Germany (CP022682.1 Homo Sapines), Brazail (CP065869.1 Homo Sapines), China (CP076025.1Duck) and South Korea (CP071594.1 Homo Sapines) at position 351 of our isolate’s sequences.

In this study there was a genetic relatedness among A9, A12 and A13 from (human, animal, food of animal origin respectively) with 94 % genetic similarity which applied by sequencing on PCR product on ica gene. On the other hand, to investigate the evolutionary relationship between the sequenced gene from this isolates and strains available in the GenBank database, it revealed that there is genetic similarity between human isolate (A9) and swine isolate in china (CP068247.1) and food isolate in Thiland (CP075508.1) with 96%. There is genetic similarity between animal isolate (A12) and human isolates in Germany (CP077916.1), USA(CP089159.1) and china(CP083728.1) with 96%. There is genetic similarity between food of animal origin isolate (A13) and human isolate in USA (CP026067.1) and duck (CP076025.1) isolate in china with 95%.

DISCUSSION

This study was designed to determine the prevalence of coagulase positive and coagulase negative staphylococcus isolates from different sources (human, animals, food of animal origin) from Ismailia governorate and their genetic relatedness to S. aureus isolates from human, animal, and food from animal origin.

Our findings were virtually identical to those of (El-Jakee et al., 2008) who used bacterial culture techniques to determine the presence of S. aureus in samples collected from unwell individuals and found 14 S. aureus strains with a 28% success rate, while the percentage of S. aureus isolates from septic wound was higher with a percentage of 45% (Kim and Hong, 2019) found that 161 of the 198 refugee health research participants volunteered to submit a nose swab. S. aureus was found in 9 (5.6%) of the 161 nasal swabs. MRSA was detected in four (2.5%) of the genotypes. These findings take into consideration research indicating that the most of hospital-acquired infections are caused by S. aureus. In this study, 2 S. aureus isolates were found in sputum from a dog with respiratory symptoms and an abscess from a cat, which corresponded with previous findings. (Lu and McEwan, 2007) who investigated the microbiota on healthy skin in 10 dogs and 10 cats. Surprisingly, they discovered S. aureus in nine (90%) of the dogs and four (40%) of the cats, but no S. intermedius isolates.

The occurrence of Staphylococcus in raw milk and raw meat may be linked to filthy worker hands, poor water quality used to wash utensils and carcasses, unsanitary transportation, improper storage, and contamination from mastitic cattle resuming to the milking practices. The current findings are consistent with the findings of (Normanno et al., 2007; De Boer et al., 2009; Ezzat et al., 2014). In this respect our results were less than the results of (Fa Mansour and Am Basha, 2009) and (Bhargava et al., 2011) who discovered that S. aureus was present in 20% and 20.5% of meat samples from different markets, respectively.

On the other hand, our results were nearly to (Goja et al., 2013) and (Ezzat et al., 2014) who retrieved S. aureus from raw beef meal in percentages of 5.55%, 12%, and 10%, respectively, and were less than the outcomes of (Kitai et al., 2005; Van Loo et al., 2007; Pu et al., 2009) that was 65%, 46% , 39.2% respectively. Therefore in this paper, the frequency of Staphylococcus recovered from unpasteurized milk is lower than the frequency of Staphylococcus recovered from pasteurized milk. (Fagundes et al., 2010) who confirmed that 7.3% of those likely Staphylococcus could transfer to consumers through milk from animals with mild mastitis. This result is noticeably lower than the findings of (Singh and Baxi, 1982) (Ezzat et al., 2014) They reported that the incidence of Staphylococcus in raw milk samples was (21%) and (18.5%). The findings of this study differ from those of previous studies undertaken by (Abou-Khadra et al., 2020) discovered that S. aureus was present in 20% of raw milk samples from Sharkia Governorate. The incidence of Staphylococcus isolates from frozen raw chickens was low, which was consistent with the findings of (De Boer et al., 2009), who reported that imported chicken goods such as frozen filets had a low frequency of Staphylococcus. These findings differed by 2.6% from those of (Ezzat et al., 2018) , who recovered S. aureus from fresh raw chicken.

The high rate of MRSA isolation in this study was considered as an extension of previous studies conducted by other investigator teams in Egypt (Ammar et al., 2016) and Bangladesh (Nusrat et al., 2015). In this study, some of virulence and resistance genes (nuc, coa, spa, ica, mecA, blacZ) were found using PCR technique, which is the most promising since it can detect just a few microbes in clinical samples yet is quick, inexpensive, and sensitive.

All negative coagulase Staphylococcus isolates showed no evidence of having virulence factors which was confirmed by PCR detection for virulence genes (nuc, coa, spa, ica). This finding is disagreed with (Xu et al., 2022) who supposed that some CoNS present these genes. In the same context (Quinn et al., 2015) Apart from a little quantity of toxins and exoenzymes, Staphylococcus species that were previously believed to be harmless members of the microbiota of people and animals, like the majority of CoNS, were shown to be deficient in virtually all of the virulence determinants outlined for S. aureus.

Furthermore, all CoNS showed no evidence of multidrug resistance, which was confirmed by PCR testing for resistance genes (mecA, blacZ), demonstrating that this kind of Staphylococcus strain has no clinical value; so, my findings are perfectly consistent with the literature (Becker et al., 2014) whic indicated that CoNS regarded both animal and human microflora as harmless invaders.

Our findings on virulence and resistance genes were substantially identical to those of another investigation by (Parth et al., 2016) who discovered that 32 of 53 isolates tested positive for the coa gene and had three unique product sizes of 723, 812, and 1000 bp. Similar results were obtained by (Himabindu et al., 2009) who revealed that the size of PCR products generated in human samples following S. aureus amplification. (Schlegelova et al., 2003) illustrated that coa gene of S. aureus isolates from lactating cows and people was found to vary in size from 650 to 1050 bp. (Reinoso et al., 2008) furthermore, PCR amplification of the S. aureus coa gene discovered in both human and cow mild mastitis and food specimens generated seven different coa types from 45 S. aureus with amplicon sizes ranging from 400 to 1000bp.The same result was obtained by (Himabindu et al., 2009) who found that isolates that were coagulase negative by tube coagulase test were coagulase positive by gene PCR assay.

In this study the prevalence of ica gene among S. aureus isolates is 69.2% which is similar to (Foroutan et al., 2022) who claimed that a total of 30 S. aureus isolates were studied to assess the influence of biofilm forming capacity on antibiotic resistance patterns, with 77.4% developing strong biofilms. The frequency of encoding biofilm genes was as follows: icaA (9.4%), icaD (9.4%), and 75%. The spa gene was detected in all S. aureus isolates (100%), which is perfectly compatible with earlier findings (Eid et al., 2018) who detected the spa locus in all strains examined. Conventional PCR methods were capable of amplifying the protein A DNA (spa) bands (Parth et al., 2016) which is a component of the cell wall of S. aureus and is covalently attached to the peptidoglycan.

The current findings suggest a significant incidence of multidrug resistant S. aureus isolates comparable to those indicated by (Quinn et al., 2015) who discovered that in areas where antibiotics can be obtained without a prescription, the development and spread of resistance is accelerated. Similarly, in nations with no clear treatment guidelines, doctors regularly over-prescribe antibiotics and the general populace over-uses them. Also, (Food and Administration, 2014)  observed that 62% of critical antibiotics for human health are marketed to food animal producers and utilized on farms, which is likely to be a significant contributor to the high incidence of multidrug resistant S. aureus isolates in animal feeds. These findings suggest that we are approaching a hazardous point when we will be unable to develop effective medications for these pathogens, (Organization, 2015) who indicated that opposition is at an all-time high throughout the world.

Every day, new resistance mechanisms emerge and spread around the world, endangering our ability to control common infectious diseases. In addition to, (Hellen et al., 2015) Drug-resistant infections have been linked to 58000 newborn sepsis fatalities in India. There is no monitoring system in Egypt that tells us how many people died as a result of the high frequency of antibiotic resistance. (Epidemiologists, 2015) confirmed that Staphylococcus infections in hospitals were virtually impossible to cure. (Cosgrove et al., 2005) methicillin resistance in S. aureus bacteremia has been linked to a significant increase in hospitalization duration and expenses. (Healy et al., 2004) tested Eight patients (47%) positive for MRSA, seven (88%) of eight patients developed septic shock, and vancomycin was used as the first line of therapy with S. aureus bacteremia. Despite this, three patients (38%) died and three acquired complications requiring long-term antibiotic treatment.(Control and Prevention, 2001) According to a compilation of information from January 1992 to June 2001, S. aureus septicemia was related with death rates ranging from 15% to 60%.

The prevalence of MRSA in this research was greater than in previous studies by (Ezzat et al., 2014) who discovered 96 patients with MRSA from 348 patients with S. aureus bacteremia, (El-Jakee et al., 2008) who demonstrated that S. aureus isolates from patients were sensitive to methicillin (Lee et al., 2001) who said that MRSA is a major problem in Korea, the rate of methicillin resistance among human S. aureus isolates in Korea is greater than 50%. In addition to, (Normanno et al., 2007; Van Loo et al., 2007; De Boer et al., 2009; Pu et al., 2009) they observed that MRSA was present in 3.75%, 2%, 11.9%, and 5% of raw beef samples, respectively. These findings contradicted the results previously reported by (Kitai et al., 2005) who found two distinct MRSA bacteria (0.5%) in 444 raw chicken meat items from Japanese retailers. In a Korean investigation that comprised 930 slaughterhouse and retail meat samples, MRSA was discovered in two chicken meat samples (0.2%) but not in any beef samples (Kwon et al., 2006). In an Italian study of 1634 food samples, six (0.4%) MRSA bacteria were found in bovine milk and cheese (Normanno et al., 2007). Many prior investigations have shown MRSA in 48.3% of mastitis cases by (Guimarães et al., 2017), 15.5% by (Wang et al., 2015), 11.6% by (Jamali et al., 2014) and 2.5% by (Moon et al., 2007). MRSA may also contaminate food and goods without altering milk according to (Parisi et al., 2016), MRSA is spread to people by mastitic milk or direct contact with dairy cows, presenting severe issues about food safety and public health.

In this study there was a genetic relatedness among A9, A12 and A13 from (human, animal, food of animal origin respectively) with 94 % genetic similarity which applied by sequencing on PCR product on ica gene. The evolutionary link between the sequenced genes from these isolates and strains accessible in the GenBank database indicated that there is 96 % genetic similarity between human isolate (A9) (On994655), swine isolate in China, and food isolate in Thailand. There is 96 % genetic similarity between sputum from dog isolate (A12) (On994656) and human isolates from Germany, the United States, and China recorded by (Decline et al., 2020). Source of MRSA transmission is due to direct contact with humans or several animals, where dogs infected with MRSA act as a reservoir and then transmit to other animals or humans. There is genetic similarity between food of animal origin isolate (A13 mastitic milk from cow) (On994657) and human isolate in USA, duck isolate in china with 95%. This result agree with (Lee, 2003) which discovered that six of the animal isolates were similar to human isolates, pointing to a probable source of human illnesses caused by ingesting contaminated food products derived from these animals, (Organization, 2015) that mentioned 1 in 5 resistant infections in humans are caused by germs from food and animals, (Frieden, 2013) recorded 22 % of antibiotic-resistant illness in humans is in fact linked to food and (Van Loo et al., 2007) who claimed that MRSA from an animal reservoir has recently invaded the human population and is currently responsible for more than 20% of all MRSA in the Netherlands. In addition to (Weese, 2010) He stated that there is no doubt that MRSA may be present in foods and that it may play a role in human disease.

CONCLUSIONS AND RECOMMENDATION

The isolation rate of Staphylococcus from examined 350 humans, animals, food specimens was 4.3%. Thirteen isolate was confirmed as S. aureus and 2 isolates were CONS. PCR analysis revealed that nuc, spa, ica, coa, mecA, and blaZ genes were detected in coagulase positive Staphylococcus examined isolates. While, no virulence or antibiotic resistance genes were detected in CONS. The multiple sequences alignments (MSA) and phylogenetic tracking of 3 selected MRSA strains from (human (A9), animal (A12), food of animal origin (A13)) proved the genetic similarity reached to 94 %, which invigorate the possibility of transmission among different sources of food, animals, and human, which causes nosocomial infections.

CONFLICT OF INTEREST

The authors have declared no conflict of interest.

NOVELTY STAEMENT

The study focused on the phylogenetic tracking and possibility of transmission of MRSA strains isolated from human, animal, food of animal origin.

AUTHOR’S CONTRIBUTION

ME, FY, and MA H supervised, designed the study and took apart in writing the manuscript. MM collected the samples, identified the recovered strains and participated in writing of the manuscript. EMA analyzed the recorded data. AW participated in molecular detection of isolates and took apart in writing the manuscript.

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