Ginekologia i Poloznictwo
ISSN 1896-3315 e-ISSN 1898-0759

Detection of Extended-Spectrum ß-Lactamases (ESBLs), TEM, SHV and CTX-M genes among Staphylococcus haemolyticus isolates from cesarean section infections

Rabeea Abdul-Jaleel Ibrahim¹, Mohammed A. Almazini², Sad S. Mahdi Al-Amara^3* and Amal Abdul-Imam Almazini^{4 *}Correspondence: Dr. Sad S. Mahdi Al-Amara, Department of Pathological analyses, College of Science, University of Basrah, Basrah, Iraq, Email:

Received: 01-Dec-2023, Manuscript No. gpmp-23-122971; Editor assigned: 04-Dec-2023, Pre QC No. P-122971; Reviewed: 15-Dec-2023, QC No. Q-122971; Revised: 21-Dec-2023, Manuscript No. R-122971; Published: 29-Dec-2023

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Introduction

Cesarean section it regarded crucial risk factor for postpartum inflammatory because of uterine skin rapture, bladder catheterization, and endometritis [1,2]. The inappropriate antimicrobial agent, or prolonged used the antibiotics may lead to bacterial resistance and increased SSI rates [3,4]. S. haemolyticus it is considered one of the predominantly Coagulase Negative staphylococci (CONS). S. haemolyticus is non-motile, non-sporulation, Gram-positive and facultative anaerobic. However, the S. haemolyticus can be growth in optimal temperature between 30-40 ℃ with the presence of O2 and 10% of NaCl [5]. S. haemolyticus regard as inhabitants of the human and animal microorganisms, has been many reports as a relationship with nosocomial pathogens because biodiversity of microbial and virulence apparatus including resistance to antibiotics [6-8].

TheS. haemolytics is the second most recurrently pathogenic of clinical nosocomial infections, especially blood cultures of patients with sepsis, onskin and soft tissue infections mainly existing as abscess, paronychia, and serious infections in different the body systems involving endocarditis, meningitis, joint prosthetic infections, peritonitis, otitis media, urinary tract infections, septicemia and it is widespread in the hospital situation also on the hands of workers in the health care [9,10]. Furthermore, the increased of infectious associated with medical implanted and devices were caused by the pathogen interpreting Multi-Drug Resistant (MDR) Profiles [11,12]. Detect of ESBLs-producing organisms is critical for infection control and nosocomial outbreak prevention. The purpose of this study was detect Extended-Spectrum ß-Lactamases (ESBLs) genes TEM, SHV and CTX-M genes inS. haemolyticus isolates from cesarean section infections from Al-Basrah governorate, Iraq.

Materials and Methods

Collection of specimens

One hundred and fifty swab samples were collected from women underwent cesarean section for both emergency cesarean delivery, and elective cesarean delivery during period from Oct. 2022 to Jan. 2023 in Al -Basrah Teaching Hospital were selected depending on their medical history.

Isolation and identification

The samples were collected from patients underwent cesarean section. The swab samples were cultivated, the samples that gave positive results for bacterial growth identified by Vitek® 2 system.

Detection of Extended Spectrum β-Lactamase (ESBLs)

Double Disk Approximation Method (DAM)

The S. haemolyticus isolates was tested for produce Extended Spectrum β-Lactamase (ESBLs)by cultured on Mueller-Hinton agar plates, with disks of 20µg amoxicillin / 10µg clavulonic acid was placed in the center of the agar surface in center, (30 µg) cefotaxime, (30 µg) ceftriaxone, (30 µg) ceftazidime, and (30 µg) aztreonam arranged around it were approximately (20-30 mm center to center). The test was positive after an overnight incubation, with an enhanced zone of inhibition between disks [13].

Detection of ESBLs genes

Three specific primers that were used for the amplification of β-lactamase genes include: TEM, SHV and CTX-M genes Tab. 1.

Tab. 1. Specific primers used for detection of extended spectrum β-lactamase genes.

Results

From Oct. 2022 to Jan. 2023, 150 swab samples was collected from cesarean section patients at Al-Basrah Teaching Hospital were gave 57(38%) positive bacterial growth, 93(62%) negative for bacterial growth, and 51(89.5%) Gram -positive bacterial isolates, while 6(10.5%) Gram-negative bacterial isolates. As shown in Tab. 2.

Tab. 2. Percentage of bacterial species isolated from cesarean section infections.

Identification of bacterial growth by using Vitek® 2 system have been emerged various bacterial species, the most predominant were S. haemolyticus, out of 57 bacterial isolates the 39(68. 42%) isolates, S. aureus 6(10.53%), Klebsiella spp 4(7.00%) isolates, Staphylococcus saprophyticus 3(5. 30%) isolates, E. coli 2 (3.50%) isolates, S. sciuri 1(1.75%) isolate, S. hominis 1(1.75%) isolate, and E. faecium 1(1.75%) isolate, as shown in Fig. 1.

Fig. 1. Frequency of bacterial species isolated from cesarean section infections.

In present study emerged that out of (n=39) isolates were distributed to 28 (82%) S. haemolyticus isolates and 6(18%) S. aureus isolates was choice. The 25 (74%) S. haemolyticus isolates and S. aureus 6 (18%) isolates were gave positive results for producing extended-spectrum ß-lactamases (ESBLs). While, 3(8. 8%) S. haemolyticus isolates were shown negative results for producing ESBLs by using of the double approximation method (DAM), as shown in Fig. 2.

Fig. 2. Double-disc approximation (DAM) test to detect ESBLs produced isolates. A) Positive result for detect ESBLs and B) Negative result for detect ESBLs.

Also the results of the current study revealed out of 28(82%) S. haemolyticus isolates and 6(18%) S. aureus isolates the 33(97.1%) isolates gave positive results for the detection of TEM and SHV gene. While the 1(2.9%) isolate was shown negative result for the detection of TEM and SHV gene as shown in Fig. 3. and Fig. 4. While the results in present study were showed that all 34(100%) Staphylococcus spp.were gave positive results for detection of the CTX-M gene as shown in Fig. 5.

Fig. 3. Agarose electrophoresis patterns of TEM gene.

Fig. 4. The PCR amplifying products of SHV gene.

Fig. 5. PCR amplified products of CTX-M gene.

Discussion

The study found that Gram-positive bacteria isolates (89.5%) had a higher prevalence than Gram-negative bacteria (10.5%), contradicting previous studies that found a higher prevalence of Gram-negative bacteria (63.6%) and Gram-positive bacteria (36.4%) [14-16], these results are not compatible with this study. Whereas, another study results showed that (68. 4%) isolates Gram-negative bacteria and (31.6%) Gram-positive [17].

This variation could be attributed to environmental factors, geographical location, education level, and patients who took antibiotics before samples [18-20]. Factors such as hormonal changes, depression, menopause, hospital-acquired infections, immunocompromised patients, and long hospital stays can also contribute to the increased risk of infections [21-23]. Antibioticresistance genes are also a significantfactor [24-26].

The Vitek®2 system is an automated instrument designed for rapid and accurate identification of most staphylococci in clinical specimens [27-29]. It has been proven effective in detecting Gram-positive cocci and Gram-negative rods, with 99% accuracy and reproducibility confirmed by multiple independent studies [29]. The Colorimetric Vitek® 2 GP card is suitable for clinical samples, and has been praised for its performance in detecting Gram-positive and Gram-negative rods [30].

The results of the present study showed that (92%) isolates were gave positive result for producing extended-spectrum ß-lactamases, which agree with study by Hassuna, et al. [31] in Egypt that found (59. 7%) of their E. coli isolates were have ability to produce ESBLs. While the present study that disagreed with the study by Pandit, et al. [32] in Nepal which found that ESBLs production was at (40. 3%). Moreover, the results of other study revealed that (90%) isolates were produced ESBLs [33]. These results resembled with results.

The results of the present study showed that (97. 1%) isolates were gave positive result for TEM and SHV genes, this results which not agreement with study of Abdi, et al. [34] who reported (15%) of SHV gene. The results of the current study emerged that (97.1%) isolates were gave positive results for SHV gene, this results was compatible with study of Karimi, et al. [35] who reported that (82%) of isolates have this gene. This result was harmonious with study of Al–Ezee, et al. [36], were found (100%), but the study of Shahid, et al. [37] who were found (20%), and the study of Mahmoud, et al. [19] were found (17%), and the study of Alipour, et al. [38] who reported that absence of SHV gene.

The current study showed that (100%) isolates were gave positive results for CTX -M gene. While the results of other studies revealed that (77. 4%) isolates were CTX-M positive results [39]. On the other hand the results of other studies revealed that (43. 8%) isolates were gave positive result for CTX - M gene [40]. This result harmonious with study of Park, et al. [41]. Also bla CTX-M was found in another study of Maleki, et al. [42] who were found (92%) and [43] (100%). These results were compatible with present results.

Conclusion

The most isolates of S. haemolyticus and S. aureus were multi drug-resistant (MDR) were detected by using Vitek®2 compact system and most isolates of S. haemolyticus and S. aureus were producing extended-spectrum ß –lactamases (ESBLs).

Disclosure

None.

Funding

None.

Authors Contribution

(A) Study Design · (B) Data Collection . (C) Statistical Analysis · (D) Data Interpretation · (E) Manuscript Preparation · (F) Literature Search · (G) No Fund Collection

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