Egypt. J. Vet. Sci., pp.

1-10

Egyptian Journal of Veterinary Sciences

https://ejvs.journals.ekb.eg/

Insights into Staphylococcus epidermidis in Farmed Nile Tilapia in

Egypt: Molecular Characterization and Antibiotic Resistance
Abdelatty M. Saleh1,2*, Alaa Eldin Eissa3, Mohamed A Ghazy1, Sarah O.
Makled4 and Ahmed L. Abdel-Mawgood1
1
Biotechnology Program, Basic and Applied Sciences Institute, Egypt-Japan University of Science and
Technology (E-JUST), Alexandria, Egypt PO box 21934.
2
Fish Processing and Biotechnology Department, Faculty of Aquatic and Fisheries Sciences, Kafrelsheikh
University, Kafrelsheikh, Egypt PO box 33516.
3
Aquatic Animal Medicine and Management Department, Faculty of Veterinary Medicine, Cairo University,
Giza, Egypt PO box 11221.
4
Oceanography Department, Faculty of Science, Alexandria University, Alexandria, Egypt PO box 21568.

Abstract

S
taphylococcus epidermidis is one of the causative agents of mass mortality in farmed Nile
tilapia (Oreochromis niloticus) in Kafr Elsheikh Governorate, Egypt. Diseased farmed Nile
tilapia were collected randomly during the fall of 2023 from farms suffering from mass
mortalities. Clinical examination revealed severe lesions in the hepatopancreas and eye of the
collected diseased fish. Based on morphological and biochemical analyses, the isolated bacteria were
suggested to be Staphylococcus sp. Five of the nine isolates were selected for 16S rRNA sequencing
based on their consistent phenotypic characterization criteria. One isolate was found to be Bacillus
rugosus based on the 16S rRNA sequencing, which is non-pathogenic and out of the scope of this
study. The four remaining isolates were found to be S. epidermidis, which have similarities to other S.
epidermidis isolates. All isolated S. epidermidis were resistant to ampicillin, erythromycin,
chloramphenicol, streptomycin, and amoxicillin; however, they were moderately susceptible to
tetracycline and were sensitive to gentamicin and vancomycin.
Keywords: Nile tilapia, S. epidermidis, 16S rRNA, Phylogenetic analysis, Antibiogram.

Introduction production. For cultured tilapia, it also produces 44%

adding 259,583 tons to the overall production,
Fish are widely recognized as a valuable source of
highlighting its significant role in Egypt's
food due to their nutritional composition, high
aquaculture industry [5]. In Egypt, fish producers
palatability, and digestibility. However, diseases
have experienced substantial losses as a result of
significantly impact the fish populations within their
"summer mortality" in tilapia which is caused by
ecosystems [1]. The World Bank estimated that
several factors [6].
diseases cause an annual economic loss of 6 billion
USD in aquaculture [2]. The prevalence of diseases Ecological studies have revealed that S.
in aquatic ecosystems is influenced by various epidermidis is present in the aquatic environment [7].
environmental factors, including infectious Therefore, monitoring for bacterial pathogens is
organisms and stressors, which contribute to the crucial to protect aquaculture economy, and for early
susceptibility of fish to diseases [3]. detection of potential contamination threats. S.
epidermidis has previously been recorded as one of
Egypt is a leading producer of Nile tilapia
the bacterial pathogens that cause disease in
accounting for more than 84% of the total production
freshwater fish and marine environments [8].
in Africa [4]. Kafr Elsheikh Governorate is a major
contributor (324,479 tons) of the country's farmed It can be challenging to determine which fish
fish production, accounting for 55% of the total pathogen is responsible for specific infection
*Corresponding authors: Abdelatty M. Saleh, E-mail: [email protected], Tel.: 01144730187
(Received 17 October 2024, accepted 26 January 2025)
DOI: 10.21608/EJVS.2025.328908.2435
©National Information and Documentation Center (NIDOC)
2 ABDELATTY M. SALEH et al.

symptoms, as many bacterial pathogens can cause normal saline solution was used to rinse the external
similar clinical signs [9]. For example, while surface of the fish, followed by spraying with 70%
infections caused by S. epidermidis, Streptococcus ethyl alcohol. Under complete aseptic conditions,
agalactiae, and Aeromonas veronii are commonly fish were dissected, and then a loopful of tissues
associated with exophthalmos and A. hydrophila or from the kidney, spleen, and hepatopancreas were
A. veronii cause congestion to the hepatopancreas, it separately streaked onto tryptic soy agar (TSA),
is important to note that these pathogens can (HiMedia, India). Inoculated agar plates were
potentially cause overlapping lesions, as supported incubated at 25 °C for 24-48 hours. Single colonies
by previous records [10, 11]. To address those were selected for purification and further
challenges, 16S rRNA sequencing has become a characterization.
cornerstone in reliable and accurate identification for
Phenotypic characterization
fast detection and diagnosis of bacterial diseases
[12]. Morphological characterizations, such as shape,
size, Gram staining, and motility tests, were
Due to the intensification of modern technologies
performed on suspected pure colonies of the isolates
in aquaculture, disease prevalence has increased, and
[17]. Biochemical identification was conducted for
new pathogenic bacteria have emerged that are
confirmation of isolates using the following tests:
detrimental to fish health. This situation previously
coagulase, catalase, oxidase, Methyl Red (MR),
led to the extensive use of antibiotics, particularly in
Vogues Proskauer (VR), citrate, urease, and nitrate
developing countries, for the prophylaxis or
reduction [18]. The hemolytic activity of bacterial
management of bacterial infections in fish; however,
isolates was assessed by inoculating an aliquot of an
this practice has significantly declined in Egypt in
overnight bacterial suspension onto blood agar plates
recent years [13].
containing 5% sterile defibrinated sheep blood,
This work aimed to assess the prevalence of S. according to the described method by Ruaro et al.
epidermidis in diseased O. niloticus collected from [19]. The plates were then incubated for 24 hours at
cultured freshwater farms. It also aimed to construct 25 °C.
a phylogenetic tree based on the 16S rRNA
Identification of the isolates using 16S rRNA
sequencing of the four isolates and test their
sequencing
antibiotic susceptibility.
DNA extraction and purification
Material and Methods
The genomic DNA was extracted using the
Collection of fish samples
described procedure by Monir et al. [20]. The
Fifty samples diseased O. niloticus, averaging isolated bacteria were sub-cultured onto TSA plates
150–200 g in weight, were randomly collected to produce a fresh overnight culture. A single colony
between September and November 2023 from ten from the subculture was inoculated into 10 mL of
different fish farms in Kafr Elsheikh Governorate tryptic soy broth (TSB) and incubated at 25 °C for 24
that experienced mass mortalities and placed in hours. The fresh culture was centrifuged at 5000 rpm
sterile bags [14]. All samples were transferred within for ten minutes. Next, 400 μl of solution I (50 mM
6 hours of collection in an icebox with crushed ice to Tris.HCl pH-8.0, 50 mM EDTA pH-8.0, 25%
the Biotechnology lab, Basic and Applied Sciences sucrose, 1 mg lysozyme) were added to the washed
Institute, Egypt-Japan University of Science and cell pellet, thoroughly mixed, and incubated at 37 °C
Technology (E-JUST). for 15 minutes. The cells were treated with 400 μl of
solution II (10 mM Tris. HCl pH 8.0, 5 mM EDTA
Clinical and postmortem examination
pH 8.0, 1% SDS, 40 µg Proteinase K) and incubated
A clinical examination was applied to the fish at 55 °C for 3 hours. The suspension was then
samples, both external signs and post-mortem centrifuged at 6,000 rpm for 10 minutes. The
examination on the diseased fish following the aqueous layer on top was carefully removed to avoid
methods described by Heil [15]. protein debris and transferred to a new microfuge
Bacterial isolation tube. To precipitate the DNA, ice-cooled ethanol was
added to the aqueous phase twice. The DNA was
Bacterial isolation was performed according to pelletized by centrifugation at 12000 rpm for 10
the methods described by Metin et al. [16]. A sterile minutes. The pellet was rinsed with 70% ethanol,

Egypt. J. Vet. Sci.

 INSIGHTS INTO Staphylococcus epidermidis IN FARMED NILE TILAPIA IN EGYPT... 3

dried, and dissolved in 100 µl of TE buffer (pH 7.6). Clinical signs of infected fish were recorded,
The quality (A260/A280) of purified PCR products including exophthalmia (pop-eye), hemorrhage, and
was measured using a Nanodrop spectrophotometer. abdominal distension. The post-mortem examination
showed congestion of the hepatopancreas and spleen,
Sequencing of 16S rRNA
and distended gall bladder (Fig. 1).
Five of the nine isolates were selected for 16S
Bacterial isolation and identification
rRNA sequencing based on their consistent
phenotypic characterization criteria. The 16S rRNA The cultural characteristics of suspected
sequencing was performed by FASMAC (Kanagawa, Staphylococcus spp. were examined, and they
Japan). Briefly, the 16S rRNA gene fragment was appeared to be white-raised, cohesive colonies on
amplified with PCR using the 27F/1492R primer. TSA.
BLASTn was used for the sequence analysis by
Prevalence of S. epidermidis
putting the sequences as queries to Genbank to
confirm the identities and closest relatives of the A total of 8 out of 50 examined fishes were
samples [21]. infected by S. epidermidis (The overall prevalence
was 16%).
Phylogenetic analysis of isolates
Characteristics of isolated Staphylococcus spp.
Multiple sequences were aligned using ClustalW
in Mega 11.0. Then, the sequences of S. epidermidis Table 2 illustrates the phenotypic characteristics
and the most closely related sequences of bacteria in and biochemical identification of Staphylococci spp.
GenBank were compared (Table 1). The Maximum recovered from internal organs of diseased Nile
Likelihood approach was implemented in Mega 11.0 tilapia. The isolates were Gram-positive, cocci, and
to infer phylogenetic relationships [22]. The non-motile. They were positive for the catalase,
confidence level in the Maximum Likelihood method urease, and nitrate reduction tests. The isolated
trees was determined by analyzing 1,000 bootstrap bacteria were coagulase negative and showed no
repetitions using the Mega software. hemolysis on sheep blood agar.

Antimicrobial susceptibility testing Confirmation of isolates by 16S rRNA sequencing

An antibiotic susceptibility test was conducted on Nine bacteria were isolated from 50 diseased Nile
isolates of S. epidermidis using the disc diffusion tilapia samples cultivated on TSA and subsequently
Kirby-Bauer technique on Mueller-Hinton agar [23]. identified through biochemical and phenotypic
The following antibiotics were tested: Ampicillin assays. Based on consistent phenotypic
(10µg), Tetracycline (30µg), Erythromycin (20µg), characterization criteria, five isolates were selected
Chloramphenicol (30µg), Gentamycin (10µg), for 16S rRNA sequencing.
Vancomycin (30µg), Streptomycin (30µg), and The results of the 16S rRNA sequencing revealed
Amoxicillin (10µg). The discs were firmly placed on that one isolate was identified as Bacillus rugosus,
the agar plates previously streaked with the test which is non-pathogenic and irrelevant to this study.
organism (0.5 McFarland) using sterile forceps and Its nucleotide sequence has been deposited in the
then incubated at 37 °C for 18-24 hours. GenBank database under the accession number
Susceptibility or resistance of the isolates to different PQ865795.
antibiotics was indicated by the appearance or non-
appearance of clear zones of inhibition, which were The remaining four sequenced isolates were
measured to the nearest millimeter using a vernier identified as S. epidermidis, and their 16S rRNA
ruler. The diameters of the zone of inhibition were gene nucleotide sequences have been submitted to
compared with the cut-off points, and the the GenBank database under the following accession
interpretations of the results were performed numbers: PP781962.1, PP781976.1, PP781979.1,
according to the recommendations of the Clinical and and PP781988.1, respectively.
Laboratory Standards Institute [24]. Phylogenetic analysis of the four sequenced S.
Results epidermidis compared to the bacterial isolates
archived in the NCBI
Clinical pathology
The phylogenetic tree (Fig. 2) was generated by
aligning the sequences of our isolates with those

Egypt. J. Vet. Sci.

4 ABDELATTY M. SALEH et al.

recorded in Table 1. It illustrates the strains of S. The data presented in table 2 align with established
epidermidis that have been isolated from a variety of literature, confirming the prevalence of S.
sources, such as fish farms, natural fisheries, and fish epidermidis in the freshwater aquatic environment
[29].
products.
The phylogenetic tree of the four S. epidermidis
The first isolate, S. epidermidis strain AS1 reveals high evolutionary similarities, despite their
(PP781962.1), has a nucleotide sequence that is isolation from different fish farms in Kafr Elsheikh
99.63% the same as the reference isolate, S. Governorate. That similarity indicates the incidence
epidermidis strain B7, which was found in Shidal, of S. epidermidis in Nile tilapia's fish farms. Most
India and has the GenBank entry number KP979596. farms were irrigated with water from the same canal
The second strain of S. epidermidis we found, AS2 used for wastewater drainage, which facilitates
reinfection and exacerbates the spread of S.
(PP781976.1), is 99.77% similar to the reference
epidermidis in the farms. Contamination from
strain of S. epidermidis, LGC 305, which was found livestock feces and feed wastes deteriorates irrigation
in the gut of a freshwater Indian loach (GenBank water quality, making it difficult to decrease the risk
accession number OQ271316.1). Two of our S. of water contamination with microbes [30]. Fish
epidermidis isolates, AS3 (PP781979.1) and AS4 hatcheries can be sources of infection for fish farms,
(PP781988.1), are 99.86% and 99.93% identical with mainly because they produce and distribute fries to
the reference strain found in Chinese fermented fish various aquaculture facilities. Without strict
biosecurity measures, pathogens like bacteria,
(GenBank entry number MH491958.1).
viruses, fungi, and parasites can be introduced and
Antimicrobial susceptibility testing spread through contaminated water, equipment, or
fish stocks [31]. S. epidermidis was one of the
All eight isolated S. epidermidis samples were common bacterial causes of mortality in tilapia fries
subjected to an antibiogram test utilizing the disc collected from hatcheries in Kafr Elsheikh
diffusion method to determine their antimicrobial Governorate [32].
susceptibility profiles. The results in Figure 3 show
that all S. epidermidis isolates were sensitive to The phylogenetic tree (Figure 2) illustrates that
gentamycin and vancomycin while being moderately the four representative bacterial strains isolated from
susceptible to tetracyclines. Additionally, all isolates Nile tilapia in this study show a significant degree of
were resistant to ampicillin, erythromycin, similarity to 33 GenBank S. epidermidis strains
chloramphenicol, streptomycin, and amoxicillin. associated with fish, their habitats, and fish products
(GenBank entry number MH491958.1). from various countries.

Discussion S. epidermidis is an opportunistic pathogen that

contributes to the mass mortality of farmed tilapia,
Nile tilapia is the most widely farmed fish in impacting the economic, nutritional, and health
Egypt, valued for its ability to tolerate fluctuations in aspects of freshwater farms. Although we did not
water physicochemical parameters and its diet of perform a bacterial challenge test using the isolates, a
phytoplankton. Nile tilapia has a tremendous previous study had performed this test on sea bream,
commercial effect and high nutritional value among and they found that a high mortality rate (63.3%) as a
fish species [25]. Intensive farming practices have result of intraperitoneal injection of S. epidermidis
increased fish susceptibility to various pathogens, [33]. It is also reported that S. epidermidis caused
such as bacteria, fungi, parasites, and viruses. infections in cultured Oncorhynchus mykiss [34].
Staphylococcus spp. indisputably leads to high Data from Table 3 show different prevalence
mortality rates in freshwater aquaculture under rates of bacteria isolated from fresh and marine fish
stressful conditions [26]. S. epidermidis bacterium species. The current study is nearly lined with a
colonizes mucous membranes and skin of humans, previous survey that recorded the most common
but when transferred into fish, it infects the spleen, bacterial pathogens of the Nile tilapia fries in Kafr
kidney, and eye, causing uni or bilateral Elsheikh Governorate, Egypt. The researchers
exophthalmia [27]. Diseased Nile tilapia of isolated S. epidermidis by 20% of 6000 tilapia fries
exophthalmia is lined with symptoms first recorded [32]. In a study conducted in Taiwan, S. epidermidis
by Kusuda and Sugiyama [28] who isolated this was recorded as the most dominant pathogenic
bacterium from the eyes of cultured Seriola species isolated from moribund tilapia with a
quinqueradiata and Pagrus major. percentage of 10% (16 cases out of 159 moribund
Based on morphological and biochemical tilapia) [35]. This percentage is slightly lower than
characteristics data, 8 bacterial isolates were that of current study. Our study is matched with a
identified as of Staphylococcus sp. Illumina-based previous report in Pakistan in which the authors
16S rRNA sequencing effectively confirmed the found out the prevalence of pathogenic bacteria in
identification of the four isolates as S. epidermidis. the collected fresh fish samples is 17.85% (15

Egypt. J. Vet. Sci.

 INSIGHTS INTO Staphylococcus epidermidis IN FARMED NILE TILAPIA IN EGYPT... 5

positive samples out of 84 collected fish) [36]. S. Conclusion

epidermidis was reported as the primary causative
Accurate identification of the causative agents of
agent of mass mortality in diseased sea bass, with a
fish pathogens is crucial for effective farm
high percentage incidence of 80% (four fish were
management. This identification is challenging
infected out of five diseased) [37].
because many bacterial pathogens can cause similar
In the current study, all isolates of S. epidermidis clinical signs. Combining clinical and biochemical
were resistant to ampicillin, erythromycin, identifications and 16S rRNA sequencing is essential
chloramphenicol, streptomycin, and amoxicillin. Our for the correct diagnosis of S. epidermidis and crucial
isolates of S. epidermidis were sensitive to to avoid misleading diagnosis. Phylogenetic analysis
gentamycin and vancomycin, and they recorded an revealed a high degree of evolutionary similarity
intermediate susceptibility with tetracycline. Our between the isolates and other reference strains,
finding agrees with Eladli et al. [38], in which suggesting that shared environmental factors. An
isolates were sensitive to gentamycin and antibiogram test should be performed before using an
vancomycin. Bacteria have evolved many defense antibiotic in aquaculture to ensure effective treatment
mechanisms against antimicrobial agents, and drug- and minimize the development of antibiotic
resistant pathogens are rising. Genome studies of S. resistance mechanisms.
epidermidis have shown that various genes offer
Acknowledgments
resistance to the bacteria against severe
environmental conditions. S. epidermidis can form The authors are extremely grateful to the
biofilm by producing extracellular polysaccharides, Egyptian Ministry of Higher Education, and
proteins, and DNA to enhance its resistance Kafrelsheikh University.
mechanism [39]. The bacteria develop resistance
Conflict of interest statement
factors through genetic recombination and acquiring
new genes. Some antibiotics can accumulate in The authors have disclosed that they do not hold
sediments and the environment, causing bacteria to any conflicts of interest related to the publication of
develop resistance to many effective antibiotics. this article.
Other factors contribute to this resistance, such as the
intensive farming of various fish species, agricultural Funding
and industrial waste entering water systems, and This research was funded by the Egyptian
insufficient government regulations on the excessive Ministry of Higher Education.
use of antibiotics in aquaculture [40].
Ethical of approval
This study did not investigate the pathogenicity
of the isolates or conduct experimental challenge Not applicable.
tests. Future research will focus on assessing the
pathogenicity of S. epidermidis, performing
histopathological examinations, and applying Koch's
postulates to establish a definitive causal relationship
between the isolates and the observed disease.

Fig.1. Unilateral exophthalmia in infected farmed Nile tilapia observed in a suspected S. epidermidis case

Egypt. J. Vet. Sci.

6 ABDELATTY M. SALEH et al.

TABLE 1. Bacterial species used for constructing the phylogenetic tree

Nature of Sample Source (host/country) Accession number

Hatcheries of tilapia Tilapia fries / Egypt MT293804.1
Fish-tank Plankton / United Kingdom AJ491709.1
Product (Filleted) Sea catfish (Arius heudeloti) / Senegal EU128487.1
Estuarine Estuarine cat fish (Mystus gulio) /India JX625992.1
Ocean water fish Puffer fish (Sphoeroides annulatus) / Mexico HM584018.1
Fish processing plant Fish processing wastewater / India KC161905.1
Ocean and Coastal fish Marine fish /India KJ459012
Fish gut Fish gut/India KR006922.1
Ngari (fermented fish) Puntius sophore fish /India KJ699166.1
Fish sauce fermentation Fish sauce/ Thailand KU132366.1
Fish kills Fish kill / Korea KP115686.1
Fish processing plant Fish processing wastewater / India KT387371.1
Fermented fish Large yellow croaker/ China KX267887.1
Shidal (Fermented fish product) Puntius sophore /India KP979596.1
Fermented fish Large yellow croaker/ China KX237940.1
Salted fish Salted fish / United Arab Emirates MF067464.1
Fermented fish Fermented fish / China MH491958.1
Fish sausage Fish sausage/ China MH915445.1
Fish Product Stinky mandarin fish / China MN867689.1
Tributaries of the Pacific Ocean Rainbow trout (Oncorhynchus mykiss) / Turkey MN923048.1
Fish pond Sciaenops ocellatus / China MT071633.1
Fish pond Sciaenops ocellatus / China MT071657.1
Farmed bivalve Perna viridis /India MT491104.1
Refrigerated Fillets Sturgeon (Acipenser) / China OK103766.1
Gut of Extensively cultured Shrimp Whiteleg shrimp (Litopenaeus vannamei)/ India OK244469.1
Fish gut from estuary Esturine fish / India ON332486.1
Fish pond Fish / India ON387515.1
Farmed Fish Gilt-head bream (Sparus aurata) / Saudi Arabia OP704017.1
Freshwater fish Gut of Loach / India OQ271316.1
Pond Shing fish Shing fish gut / Bangladesh OR871836.1
Freshwater fish Blood of Cyprinus carpio koi / India MN515417.1
Farmed Nile tilapia Spleen of Oreochromis niloticus / Egypt MN153038.2
Estuarine fish Gut of Mugil jerdoni / India KJ623584.1

TABLE 2. Morpho-chemical characteristics of S. epidermidis

Characters Result
Gram stain +
Motility -
Shape Cocci
Coagulase -
Catalase +
Oxidase -
Methyl Red (MR) -
Voges Proskauer (VR) -
Citrate -
Urease +
Nitrate Reduction +
Hemolysis -
(+) Positive reaction, (-): Negative reaction

TABLE 3. Summary of the prevalence of S. epidermidis in some previous studies compared to the current study

Fish Name Percentage of Prevalence (%) Reference

Cultured Nile tilapia 16 This study
Cultured Nile tilapia fries 11.1 [32]
Cultured Tilapia 10 [35]
Cultured Silver carp 17.85 [36]
Cultured European sea bass 80 [37]

Egypt. J. Vet. Sci.

 INSIGHTS INTO Staphylococcus epidermidis IN FARMED NILE TILAPIA IN EGYPT... 7

(44) AJ491709.1 Staphylococcus epidermidis strain P5

(42)
KJ459012.1 Staphylococcus epidermidis strain AA3
(51)
EU128487.1 Staphylococcus epidermidis strain CWBI B1433
(49)
MF067464.1 Staphylococcus epidermidis strain Unknown2
(52)
ON387515.1 Staphylococcus epidermidis strain anjacbact2021a
(48) HM584018.1 Staphylococcus epidermidis strain CAIM 852
OK244469.1 Staphylococcus epidermidis strain FML3
(62)
(46) PP781976.1 Staphylococcus epidermidis strain AS2
(45) OQ271316.1 Staphylococcus epidermidis strain LGC 305
(67)
MN515417.1 Staphylococcus epidermidis strain NRLFFD301
KX237940.1 Staphylococcus epidermidis strain TM2-9
MT071657.1 Staphylococcus epidermidis strain ZJH
(72)
(70)
(59)
PP781979.1 Staphylococcus epidermidis strain AS3
(65)
PP781988.1 Staphylococcus epidermidis strain AS4
(68) MH491958.1 Staphylococcus epidermidis strain 3-9
(58)
MN867689.1 Staphylococcus epidermidis strain SE3-11
(69)
KT387371.1 Staphylococcus epidermidis strain US 108
(60)
(71) KJ623584.1 Staphylococcus epidermidis strain MJMG8.1
(66) KJ699166.1 Staphylococcus epidermidis strain KB41
MT071633.1 Staphylococcus epidermidis strain ZJH
(55) KX267887.1 Staphylococcus epidermidis strain SM5-7
OK103766.1 Staphylococcus epidermidis strain LT-03
(57) JX625992.1 Staphylococcus epidermidis strain SubaKolSe6
(64)
(53)
KU132366.1 Staphylococcus epidermidis strain M5B-P10
(56) MT491104.1 Staphylococcus epidermidis strain CMFRI/StE-05
(61)
PP781962.1 Staphylococcus epidermidis strain AS1
(54) (63) KP979596.1 Staphylococcus epidermidis strain B7
MH915445.1 Staphylococcus epidermidis strain UMBR4133
KP115686.1 Staphylococcus epidermidis strain SFCFD20130614-4
(47)
KR006922.1 Staphylococcus epidermidis strain J50
(43)
OP704017.1 Staphylococcus epidermidis strain SA7
(50) MN153038.2 Staphylococcus epidermidis strain CU-FVM-DFD-SP1
MT293804.1 Staphylococcus epidermidis strain CU-FVM-AAM-TSH3
OR871836.1 Staphylococcus epidermidis strain CFS13
(40)

(41)
ON332486.1 Staphylococcus epidermidis strain MH-JMC16
(38)
KC161905.1 Staphylococcus epidermidis strain CL1108
(39) MN923048.1 Staphylococcus epidermidis strain VSUF1

Fig.2. Phylogenetic tree of isolates compared to recorded isolates showing Evolutionary analysis

Phylogenetic tree of chosen members of S. epidermidis based on 16S rRNA sequences. The GenBank accession numbers for
every sequence that constitutes the analysis are provided before the taxon names. The red color shows the sequences found in
this study. The evolutionary history was inferred using the Maximum Likelihood method and Hasegawa-Kishino-Yano
model. The tree with the highest log likelihood (-7703.26) is shown. Initial tree(s) for the heuristic search were obtained
automatically by applying Neighbor-Join and BioNJ algorithms to a matrix of pairwise distances estimated using the
Maximum Composite Likelihood (MCL) approach, and then selecting the topology with superior log likelihood value. A
discrete Gamma distribution was used to model evolutionary rate differences among sites (5 categories (+G, parameter =
1.9541)). This analysis involved 37 nucleotide sequences. There were a total of 1996 positions in the final dataset.

Egypt. J. Vet. Sci.

8 ABDELATTY M. SALEH et al.

25

20
Mean Inhibition Zone (mm)

15 Isolate 1
Isolate 2
Isolate 3
10
Isolate 4
Isolate 5
Isolate 6
5
Isolate 7
Isolate 8
0

Antibiotics

Fig.3. Antibiogram test for S. epidermidis against all the tested antibiotics under study
R: Resistant (0-12 mm), I: Intermediate (13- 17 mm), S: Susceptible (≥17 mm)

Wild Fish, Sixth Edition. Praxis Publishing,

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‫ في أسمبك البلطي النيلي‬Staphylococcus epidermidis ‫رؤي حىل بكتيريب‬

‫ التىصيف الجسيئي ومقبومة المضبدات الحيىية‬:‫المستسرعو في مصر‬
2
‫ و أحمذ لطفً عبذالمىجىد‬4‫ سبره أسبمو مقلذ‬، 2‫ دمحم عبذالهبدي غبزي‬، 3ً‫ عالء الذين عيس‬،*2،1‫عبذالعبطً دمحم صبلح‬
.‫ يصز‬،‫ اندايؼّ انًصزيّ انيابايُّ نهؼهٕو ٔانخكُٕٔنٕخيا‬،‫ يؼٓذ انؼهٕو األساسيّ ٔانخطبيقيت‬،ٗ‫ بزَايح انبيٕحكُٕنٕخ‬1
.‫ يصز‬،‫ خايؼت كفز انشيخ‬،‫ كهيت ػهٕو انثزٔة انسًكيّ ٔانًصايذ‬،ٗ‫ قسى حصُيغ األسًاك ٔانبيٕحكُٕنٕخ‬2
.‫ يصز‬،ِ‫ خايؼت انقاْز‬،ٖ‫ كهيت انطب انبيطز‬،‫ قسى طب ٔرػايّ األحياء انًائيت‬3
.‫ يصز‬،‫ خايؼت األسكُذريت‬،‫ كهيت انؼهٕو‬،‫ قسى ػهٕو انبحار‬4

‫الملخص‬
ّ‫ أحذ انؼٕايم انًسببت نهُفٕق اندًاػي في أسًاك انبهطي انُيهي انًسخزرػ‬Staphylococcus epidermidis ‫ح َؼذُّ بكخيزيا‬
‫ يٍ أسًاك انبهطي انُيهي انًصابّ يٍ انًزارع‬2223 ‫ حى خًغ ػيُاث ػشٕائيت فٗ خزيف‬.‫ يصز‬،‫في يحافظت كفز انشيخ‬
ٍ‫ حيث كشفج انفحٕصاث االإلكهيُيكيت ػٍ ٔخٕد إصاباث شذيذة في كبذ ٔػي‬، ‫انخي حؼاَي يٍ حاالث َفٕق خًاػي‬
‫ ٔباسخخذاو انفحٕصاث انًٕرٔفٕنٕخيّ انظاْزيّ ٔانبيٕكيًيائيت حبيٍ أٌ انبكخيزيا‬،‫األسًاك انًزيضّ انخٗ حى خًؼٓا‬
، ‫انًؼزٔنت حُخًٗ إنٗ انًكٕراث انؼُقٕديت‬

16S ‫حى اخخيار خًست يٍ بيٍ حسؼت ػزالث بُاء ػهٗ حشابّ انفحٕصاث انًٕرٔفٕنٕخيّ انظاْزيّ ٔانبيٕكيًيائيت نؼًم‬
‫ ْٔي بكخيزيا غيز يًزضت ٔخارج‬Bacillus rugosus ‫ ٔأظٓزث انُخائح أٌ إحذٖ انؼزالث‬،rRNA sequencing
‫ طبقا نهخشابّ انديُٗ يغ‬S. epidermidis ‫ بيًُا انؼزالث األربؼّ انًخبقيت فقذ حى حؼزيفٓى كبكخيزيا‬،‫َطاق ْذِ انذراست‬
.‫انؼزالث انًزخؼيت انًسخخذيت‬

،‫ انكهٕرايفيُيكٕل‬،ٍ‫ اإلريثزٔييسي‬،ٍ‫ يقأيت نأليبيسهي‬S. epidermidis ‫أظٓزث خًيغ انسالالث انًؼزٔنت نبكخيزيا‬
ّ‫ بيًُا كاَج انبكخيزيا أكثز حساسي‬، ٍ‫ ٔأظٓزٔا حساسيّ يخٕسطّ نهخخزاسيكهي‬،ٍ‫ ٔاأليٕكسيسيهي‬،ٍ‫انسخزبخٕيايسي‬
.ٍ‫نهدُخاييسيٍ ٔانفاَكٕيايسي‬
.ّ‫ حساسيت انًضاداث انحيٕي‬، ‫ ححهيم انخطٕر‬، 16S rRNA ،ّ‫ انؼُقٕديّ انبشزٔي‬، ‫ انبهطي انُيهي‬:‫الكلمبت الذالة‬

Egypt. J. Vet. Sci.