|Year : 2021 | Volume
| Issue : 2 | Page : 75-80
Characterization and antibiotic resistance of staphylococci strains isolated from brack hospital in the Southern Region of Libya
Ghada M Alshaibani1, Aisha M Shahlol2, Ahmed S Abid3, Samira G Amri4, Ezzeddin Sh Aghila4, Ghaith A Abdalah4, Mohamed Omar Ahmed5
1 Department of Life Sciences, School of Basic Sciences, Libyan Academy of Postgraduate Studies, Tripoli, Libya
2 Department of Medical Laboratory Technology, Faculty of Engineering and Technology, Sabha University, Sabha, Libya
3 National Centre for Disease Control, Tripoli, Libya
4 Burns and Plastic Surgery Centre, Tripoli, Libya
5 Department of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Tripoli, Tripoli, Libya
|Date of Submission||30-Jun-2021|
|Date of Decision||06-Aug-2021|
|Date of Acceptance||12-Aug-2021|
|Date of Web Publication||6-Jan-2022|
Mohamed Omar Ahmed
Department of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Tripoli, P. O. Box: 13662, Tripoli
Source of Support: None, Conflict of Interest: None
Background: Staphylococcus species are one of the most important healthcare-associated pathogens able to acquire and spread multidrug-resistant determinants. Objectives: The present study investigated and analyzed 110 potential Staphylococcus species isolated from various clinical and nonclinical samples at Brack hospital. Materials and Methods: A nonselective laboratory approach was applied, using cultural characteristics, Gram stain, and catalase reactivity followed by confirmation at the species levels and determination the susceptibility against antimicrobial agents using the Phoenix automated microbiological system. Results: In total, 57.5% were confirmed as species and subspecies of Staphylococcus represented by ten different species: nine subspecies of coagulase-negative staphylococci (CoNS) (76.2%) and one coagulase-positive staphylococci (CoNS) subspecies (23.8%). Of these strains, 16.6% were identified as methicillin-resistant staphylococci (MRS) mostly of the CoNS group expressing significant resistance to important antimicrobial classes. Conclusion: This study reports a high prevalence of various staphylococci species, particularly of CoNS group expressing multidrug resistance patterns of public health concern, from a healthcare setting in the south region of Libya. The identification of higher rate of MRCoNS underlines the importance of monitoring all multidrug-resistant staphylococci species requiring further epidemiological investigations.
Keywords: Antimicrobial susceptibility testing, Brack hospital, coagulase-negative staphylococci, Libya, multidrug-resistant staphylococci
|How to cite this article:|
Alshaibani GM, Shahlol AM, Abid AS, Amri SG, Aghila ES, Abdalah GA, Ahmed MO. Characterization and antibiotic resistance of staphylococci strains isolated from brack hospital in the Southern Region of Libya. Libyan Int Med Univ J 2021;6:75-80
|How to cite this URL:|
Alshaibani GM, Shahlol AM, Abid AS, Amri SG, Aghila ES, Abdalah GA, Ahmed MO. Characterization and antibiotic resistance of staphylococci strains isolated from brack hospital in the Southern Region of Libya. Libyan Int Med Univ J [serial online] 2021 [cited 2022 Dec 9];6:75-80. Available from: https://journal.limu.edu.ly/text.asp?2021/6/2/75/335051
| Introduction|| |
Staphylococcus species are major healthcare-associated pathogens responsible for critical and opportunistic infections among humans associated with increased morbidity and mortality rates. Methicillin-resistant staphylococci (MRS) are bacterial pathogens expressed by variable Staphylococcus species showing significant multidrug resistance to important antimicrobial classes including drugs of last resort such as glycopeptides. MRS represent serious medical and public health concerns that rapidly spread with variable epidemiological distribution worldwide.
In Libya, methicillin-resistant Staphylococcus aureus (MRSA) is the most reported nosocomial pathogen exclusively isolated from human healthcare settings; however, most of the available data are inadequate, and mainly originated from urban areas (i.e., cities from the north coastal region of Libya) with a paucity of information from suburban and underdeveloped areas., From the south region of Libya, only two studies have documented healthcare-associated organisms (i.e., MRSA and Pseudomonas aeruginosa) isolated at Sebha medical center., Recently, MRS have been reported from humans and companion animals, mainly belonging to the species of coagulase-negative staphylococci (CoNS) presenting public health and zoonotic concerns., The current study investigated a collection of 110 identified staphylococci strains isolated from humans from various clinical and nonclinical sources at Brack hospital in the southern region of Libya between August and December 2018. The collected isolates were further analyzed to identify and confirm the genus and species of the isolates and further characterize the antimicrobial susceptibility profiling. Brack hospital is a major healthcare setting that provides various medical and health services to the community in the southern region of Libya.
| Materials and Methods|| |
Source of the collection
The collection was isolated from patients (n = 63) and healthcare works (n = 47) and obtained from nasal swabs (n = 23), hand swaps (n = 73), and clinical samples from urine catheters (n = 14). The collection originated from 79% (n = 87) of females and 21% (n = 23) of males, and the age of the involved individuals ranged from 1.5 to 63 years (mean = 28.5 years). Isolates were identified based on standard laboratory procedures using direct culturing onto blood and MacConkey agar plates and further identified based on the typical criteria of staphylococci species including Gram-positive cocci, clustering, and catalase reactivity. Isolates were stored at −20°C until further analysis.
Laboratory identification and biochemical characterization
Each isolate was enriched in brain heart enrichment broth and incubated aerobically for 24–48h at 37°C. A loopful from each broth was streaked onto both mannitol salt and blood agar and incubated for 24h at 35°C. Plates were then checked for typical growing colonies featuring staphylococci as yellow, circular, and shiny colonies. A typical colony was selected from each plate and further examined with a Gram stain and catalase test and identified as presumptive staphylococci. Isolates were further tested with a BD Phoenix automated identification and susceptibility testing system (PAMS, MSBD Biosciences, Sparks, MD, USA) for definite characterization at the genus and species levels and to determine the susceptibility against antimicrobial agents. The antimicrobial susceptibility profile was identified based on the interpretation of the Phoenix system and by the criteria of CLSI guidelines. The detection of MRS was based on the interpreted criteria of minimum inhibitory concentration (MIC) for oxacillin and cefoxitin as follows: Susceptible, MIC ≤2 μg/ml, and resistant, MIC ≥4 μg/ml. The MIC breakpoints for CoNS (excluding S. lugdunensis) and S. pseudintermedius were ≤0.25 mcg/ml for susceptibility and ≥0.5 mcg/ml for resistance.
| Results|| |
In total, only 57.5% (n = 42 of 73) were confirmed as species and subspecies represented by ten different staphylococci species: nine species of a space after CoNS. (n = 32; 76.2%) and one subspecies of (CoPS) (n = 10; 23.8%) [Table 1]. The identified species were S. aureus (n = 10), S. gallinarum (n = 6), S. xylosus (n = 6), S. saprophyticus (n = 4), 2; S. epidermidis (n = 4), S. warneri (n = 4), S. equorum (n = 2), S. simulans (n = 2), S. kloosii (n = 2), and S. hominis (n = 2), [Table 1]. Furthermore, 16.6% (n = 7 out of 42; 6 CoNS and 1 CoPS) of the strains expressing MRS phenotypes were found to have similar antibiogram profiles [Table 2]. The remaining isolates were susceptible to all antimicrobial classes.
|Table 2: Antimicrobial susceptibility profiling of methicillin-resistant staphylococci expressing strains (n=7)|
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The seven MRS isolates were respectively distributed between S. gallinarum (n = 3), S. xylosus (n = 2), S. aureus (n = 1), and S. equorum (n = 1). Of these, five MRS isolates expressed typical resistance to penicillin, oxacillin, ampicillin, amoxicillin–clavulanate, cefoxitin, and cefotaxime but were susceptible to gentamicin, tetracycline (except one), trimethoprim–sulfamethoxazole, nitrofurantoin, moxifloxacin, rifampin, ciprofloxacin, linezolid, daptomycin, teicoplanin, vancomycin, fusidic acid, erythromycin, clindamycin, and mupirocin. Two MRS S. xylosus isolates expressed MLSBi phenotype expressing further resistance to erythromycin, clindamycin, trimethoprim, and tetracycline [Table 2].
| Discussion|| |
In general, most of the available information on Staphylococcus from Libya has focused on S. aureus of clinical sources with little information on other species. A previous study investigated a collection of 218 isolates of staphylococci originated from clinical samples collected at Tripoli hospital reported MRSA in 28.4% (n = 62/218) of the collection followed by MRCoNS (21.5%; n = 47/218). Another study involving different hospitals in Benghazi reported an MRSA prevalence at 8% of samples collected from various surfaces and environmental sources and identified 32 out of 100 S. aureus strains expressing the MRS phenotype but susceptible to vancomycin and mupirocin.
In the current study, 42 isolates were confirmed as Staphylococcus species of which seven isolates expressed MRS phenotypes predominantly of the CoNS group originating from nasal and hand samples. A previous study from the southern region of Libya investigating 43 strains of S. aureus recovered from different departments at Sebha medical center reported MRSA in 16% of the isolates but susceptible to vancomycin. The revelation of the present study showed the variable epidemiological status of Staphylococcus species in the studied area but may also indicate an epidemiological shift in the distribution of Staphylococcus species within the Libya healthcare system. In fact, a recent molecular investigation on a collection of clinical S. aureus collected at the largest Libyan hospital in Tripoli revealed the presence of atypical genotypic strains among MRSA strains showing a dynamic molecular shift in MRSA consistent with global molecular changes. Nevertheless, the available information on the epidemiological distribution of Staphylococcus species within the Libyan health system and the community is incomplete and inadequate mainly due to underdeveloped infrastructures and economic resources.
S. aureus and S. epidermidis are typical colonizers of the skin and nares, linked to biofilm formation and responsible for serious persistent infections., CoNS, on the other hand, are emerging opportunistic organisms able to persist on a variety of environmental surfaces but less involved in community-associated diseases. The majority of the CoNS species identified in the current study are typical human-associated organisms (i.e., the S. epidermidis-like group – S. epidermidis, S. haemolyticus S. capitis, S. hominis) and more commonly isolated from opportunistic and bloodstream infections associated with antibiotic use and the use of medical devices.,
Staphylococci of animal and farms origins are frequently associated with human and bloodstream infections showing multidrug resistant to important antimicrobial classes (e.g., glycopeptides and fluoroquinolones)., Of these, S. gallinarum, known to be widespread in the environment and isolated from skin and respiratory tract of farm animals, is increasingly reported from opportunistic infections associated with the prevention use of antibiotics. S. xylosus is a commensal organism of the skin and the mucous membranes of humans and animals responsible for opportunistic and zoonotic infections (e.g., mastitis or dermatitis). This species is frequently isolated from animal products (e.g., cheese, milk and meat) and used in the development of flavor and food processing due to its antioxidant and degradation properties. The identification of such animal- and environmental-associated species raises concerns over zoonotic contamination and contacts.
MRSA and MRCoNS are commonly isolated from humans and animals responsible for severe infections in healthcare facilities and the community., MRCoNS are an emerging cause of hospital-acquired infections; however, the available knowledge on their prevalence is very limited from the underdeveloped regions., In the current study, six MRCoNS were identified showing similar multidrug-resistance properties (i.e., AMP, Pen, AMX, OXA, FOX, CTX) of which three were S. gallinarum and expressed further and variable intermediate susceptibility to erythromycin, clindamycin, and teicoplanin. In addition, two MR S. xylosus strains were characterized expressing MLSB phenotype showing further resistance to trimethoprim–sulfamethoxazole and tetracycline which reportedly linked to antibiotics usage.
Nasal/nasopharyngeal colonization with MRCoNS is documented as a major risk factor for persistent and drug-resistant infections., CoNS are recognized as a major reservoir of virulent and antibiotic resistance genes that can be acquired by other staphylococci mainly through the transconjugant transfer of the staphylococcal cassette chromosome mec (SCCmec) transposon containing the mecA gene, as in the case of transfer between S. aureus and S. epidermidis. Another mec gene homolog is mecC, which has about 70% comparability with the mecA gene, and can be carried by SCCmec elements and isolated from animals, human clinical specimens, and the environment. Reportedly, the identification of the high rate of MRCoNS, particularly those carrying the mec genes among human isolates, may reflect higher exposure to antimicrobial drugs and/or the coexistence of resistance determinants. This may favor the horizontal transfer of mobile genetic elements to other commensal organisms leading to the emergence of more virulent drug-resistant strains such as vancomycin-resistant S. aureus., Unfortunately, due to the limitations of the current study, these important genes (i.e., mecA, mecC, and pvl) were not investigated.
As the global epidemiological distribution of staphylococci and the associated multidrug resistance phenotypes have dramatically changed, accurate laboratory identification and characterization is important., The Phoenix system proved to be efficient for accurate identification and antimicrobial susceptibility of staphylococci including CoNS as well as other major healthcare-associated pathogens such as enterococci and Gram-negative rods. Strains of S. aureus may also be misinterpreted using phenotypic testing methods and microbiological automated systems with other CoPS or certain clinically important CoNS strains such as S. lugdunensis and S. schleiferi. Therefore, reliable, efficient, and advanced molecular laboratory methods such as MALDI-TOF MS or polymerase chain reaction technology are required.
| Conclusion|| |
This study revealed novel information on important healthcare-associated pathogens isolated from a healthcare setting in the southern region of Libya. The current investigation revealed a high rate of CoNS among the collection expressing concerning MRS phenotype underlying the importance of monitoring all Staphylococcus species. Appropriate and effective prevention strategies in healthcare facilities, including antibiotics stewardship and epidemiological studies, are required to control the dissemination of MRS.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Littmann J, Viens AM. The ethical significance of antimicrobial resistance. Public Health Ethics 2015;8:209-24.
Ahmed MO, Baptiste KE, Daw MA, Elramalli AK, Abouzeed YM, Petersen A. Spa typing and identification of pvl genes of meticillin-resistant Staphylococcus aureus
isolated from a Libyan hospital in Tripoli. J Glob Antimicrob Resist 2017;10:179-81.
Ahmed MO, Elramalli AK, Baptiste KE, Daw MA, Zorgani A, Brouwer E, et al.
Whole Genome Sequence Analysis of the First Vancomycin-Resistant Enterococcus faecium
Isolates from a Libyan Hospital in Tripoli. Microb Drug Resist 2020;26:1390-8.
Ahmad KM, Alamen AA, Atiya FA, Elzen AA. Prevalence of methicillin-resistant Staphylococcus aureus
(MRSA) among Staphylococcus aureus
collection at Sebha medical center. J Adv Lab Res Biol 2018;9:01-8.
Ahmad KM, Alamen AA, Shamsi SA, Elzen AA. Co-expression of extended spectrum β-lactamase (ESBL) and AmpC β-lactamase among Pseudomonas aeruginosa
clinical isolates in sebha medical center. J Pure Appl Sci 2019;18:11-6.
Elnageh HR, Hiblu MA, Abbassi MS, Abouzeed YM, Ahmed MO. Prevalence and antimicrobial resistance of Staphylococcus
species isolated from cats and dogs. Open Vet J 2021;10:452-6.
Othman AA, Hiblu MA, Abbassi MS, Abouzeed YM, Ahmed MO. Nasal colonization and antibiotic resistance patterns of Staphylococcus
species isolated from healthy horses in Tripoli, Libya. J Equine Sci 2021;32:61-5.
Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing. 30th
ed. Wayne, Pennsylvania: CLSI Supplement M100; 2020.
Aetrugh SM, Aboshkiwa MA, Husien WM, Erhuma ME, Corrente M, Grandolfo E, et al.
Antimicrobial resistance profile and molecular characterization of methicillin-resistant staphylococcus
isolates in Tripoli Central Hospital, Libya. Libyan Int Med Univ J 2017;2:74-83. [Full text]
Al-Abdli NE, Baiu SH. Isolation of MRSA Strains from hospital environment in Benghazi city, Libya. Am J Inf Dis Microbiol 2016;4:41-3.
Otto M. Staphylococcus
epidermidis--the 'accidental' pathogen. Nat Rev Microbiol 2009;7:555-67.
Schilcher K, Horswill AR. Staphylococcal biofilm development: Structure, regulation, and treatment strategies. Microbiol Mol Biol Rev 2020;84:e00026-19.
Becker K, Heilmann C, Peters G. Coagulase-negative staphylococci. Clin Microbiol Rev 2014;27:870-926.
Mendoza-Olazarán S, Morfin-Otero R, Rodríguez-Noriega E, Llaca-Díaz J, Flores-Treviño S, González-González GM, et al.
Microbiological and molecular characterization of Staphylococcus hominis isolates from blood. PLoS One 2013;8:e61161.
Nobrega DB, Naushad S, Naqvi SA, Condas LA, Saini V, Kastelic JP, et al.
Prevalence and genetic basis of antimicrobial resistance in non-aureus
staphylococci isolated from Canadian dairy herds. Front Microbiol 2018;9:256.
Velasco V, Buyukcangaz E, Sherwood JS, Stepan RM, Koslofsky RJ, Logue CM. Characterization of Staphylococcus aureus
from Humans and a comparison with İsolates of animal origin, in north Dakota, United States. PLoS One 2015;10:e0140497.
Giordano N, Corallo C, Miracco C, Papakostas P, Montella A, Figura N, et al.
Erythema nodosum associated with Staphylococcus xylosus
septicemia. J Microbiol Immunol Infect 2016;49:134-7.
Leroy S, Vermassen A, Ras G, Talon R. Insight into the genome of Staphylococcus xylosus
, a ubiquitous species well adapted to meat products. Microorganisms 2017;5:E52.
Bal EB, Bayar S, Bal MA. Antimicrobial susceptibilities of coagulase-negative staphylococci (CNS) and streptococci from bovine subclinical mastitis cases. J Microbiol 2010;48:267-74.
Lakhundi S, Zhang K. Methicillin-resistant Staphylococcus aureus
: Molecular characterization, evolution, and epidemiology. Clin Microbiol Rev 2018;31:e00020-18.
Venugopal N, Mitra S, Tewari R, Ganaie F, Shome R, Rahman H, et al.
Molecular detection and typing of methicillin-resistant Staphylococcus aureus
and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India. Vet World 2019;12:1760-8.
Morgenstern M, Erichsen C, Hackl S, Mily J, Militz M, Friederichs J, et al.
Antibiotic resistance of commensal Staphylococcus aureus
and coagulase-negative Staphylococci
in an International cohort of surgeons: A prospective point-prevalence study. PLoS One 2016;11:e0148437.
Han JI, Yang CH, Park HM. Prevalence and risk factors of Staphylococcus
spp. carriage among dogs and their owners: A cross-sectional study. Vet J 2016;212:15-21.
Kateete DP, Asiimwe BB, Mayanja R, Mujuni B, Bwanga F, Najjuka CF, et al.
Nasopharyngeal carriage, spa types and antibiotic susceptibility profiles of Staphylococcus aureus
from healthy children less than 5 years in Eastern Uganda. BMC Infect Dis 2019;19:1023.
Xu Z, Shah HN, Misra R, Chen J, Zhang W, Liu Y, et al.
The prevalence, antibiotic resistance and mecA
characterization of coagulase negative staphylococci recovered from non-healthcare settings in London, UK. Antimicrob Resist Infect Control 2018;7:73.
Bietrix J, Kolenda C, Sapin A, Haenni M, Madec JY, Bes M, et al.
Persistence and Diffusion of mec
C-positive CC130 MRSA isolates in dairy farms in Meurthe-et-Moselle county (France). Front Microbiol 2019;10:47.
McGuinness WA, Malachowa N, DeLeo FR. Vancomycin resistance in Staphylococcus aureus.
Yale J Biol Med 2017;90:269-81.
Gómez-Sanz E, Ceballos S, Ruiz-Ripa L, Zarazaga M, Torres C. clonally diverse Methicillin and multidrug resistant coagulase negative staphylococci are ubiquitous and pose transfer ability between pets and their owners. Front Microbiol 2019;10:485.
Sunagar R, Hegde NR, Archana GJ, Sinha AY, Nagamani K, Isloor S. Prevalence and genotype distribution of methicillin-resistant Staphylococcus aureus
(MRSA) in India. J Glob Antimicrob Resist 2016;7:46-52.
Carroll KC, Borek AP, Burger C, Glanz B, Bhally H, Henciak S, et al.
Evaluation of the BD Phoenix automated microbiology system for identification and antimicrobial susceptibility testing of staphylococci and enterococci. J Clin Microbiol 2006;44:2072-7.
Chatzigeorgiou KS, Sergentanis TN, Tsiodras S, Hamodrakas SJ, Bagos PG. Phoenix100 versus Vitek 2 in the identification of gram-positive and gram-negative bacteria: A comprehensive meta-analysis. J Clin Microbiol 2011;49:3284-91.
Guardabassi L, Damborg P, Stamm I, Kopp PA, Broens EM, Toutain PL, et al.
Diagnostic microbiology in veterinary dermatology: Present and future. Vet Dermatol 2017;28:146-e30.
[Table 1], [Table 2]