Search

Background: The purpose of this study was to look into the presence of plasmid-mediated quinolone resistance (PMQR) genes and biofilm formation in several species of clinical Shigella isolates that were resistant to quinolones. Methods: The stool samples of 150 patients (younger than 10 years) with diarrhea were collected in this cross-sectional study (November 2020 to December 2021). After cultivation of samples on Hektoen Enteric agar and xylose lysine deoxycholate agar, standard microbiology tests, VITEK 2 system, and polymerase chain reaction (PCR) were utilized to identify Shigella isolates. The broth microdilution method was used to determine antibiotic susceptibility. PMQR genes including qnrA, qnrB, qnrC, qnrD, qnrE, qnrS, qnrVC, qepA, oqxAB, aac(6′)-Ib-cr, and crpP and biofilm formation were investigated in quinolone-resistant isolates by PCR and microtiter plate method, respectively. An enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique was used to determine the clonal relatedness of quinolone-resistant isolates. Results: A total of 95 Shigella isolates including S. sonnei (53, 55.8%), S. flexneri (39, 41.1%), and S. boydii (3, 3.2%) were identified. The highest resistance rates of the isolates were against ampicillin (92.6%, n = 88/95). Overall, 42 of 95 (44.2%) isolates were simultaneously resistant against two or more quinolones including 26 (61.9%) S. sonnei and 16 (38.1%) S. flexneri. All isolates were multidrug-resistant (resistance to more than 3 antibiotics). The occurrence of PMQR genes was as follows: qnrS (52.4%), qnrA and aac(6′)-Ib-cr (33.3%), and qnrB (19.0%). The prevalence in species was as follows: 61.5% and 37.5% (qnrS), 19.2% and 56.3% (qnrA), 38.5% and 25.0 (aac(6′)-Ib-cr), and 19.2% and 18.8% (qnrB) for S. sonnei and S. flexneri, respectively. The other PMQR genes were not detected. In total, 52.8% (28/53) of quinolone-susceptible and 64.3% (27/42) of quinolone-resistant isolates were biofilm producers. Biofilm formation was not significantly different between quinolone-resistant and quinolone-susceptible isolates (P-value = 0.299). Quinolone-resistant isolates showed a high genetic diversity according to the ERIC-PCR. Conclusion: It seems that qnrS, qnrA, and aac(6′)-Ib-cr play a significant role in the quinolone resistance among Shigella isolates in our region. Also the quinolone-resistant S. flexneri and S. sonnei isolates had a high genetic diversity. Hence, antibiotic therapy needs to be routinely revised based on the surveillance findings. [ABSTRACT FROM AUTHOR]

Summary: The study aimed to assess Campylobacter jejuni prevalence in chicken meat, biofilm formation, virulence factors, antibiotic resistance, and molecular typing. In the study, 200 chicken meat samples were collected from local outlets and 51 (25.5%) isolates were identified as C. jejuni. Resistance rates to ampicillin, tetracycline, sulfamethoxazole/trimethoprim, and ciprofloxacin were 59%, 60%, 64%, and 64% respectively. Many of the isolates (49%) exhibited multidrug resistance. Beta‐lactamase and tetracycline resistance genes were found in 82.3% and 86.2% of isolates, respectively. Virulence genes were detected in various proportions. Biofilm formation was weak to moderate. ERIC‐PCR showed varied band profiles. Whole genome sequencing confirmed findings. The study highlights C. jejuni presence with antibiotic resistance, virulence and biofilm features in chicken meat, posing public health risks. [ABSTRACT FROM AUTHOR]

Abstract Background The purpose of this study was to look into the presence of plasmid-mediated quinolone resistance (PMQR) genes and biofilm formation in several species of clinical Shigella isolates that were resistant to quinolones. Methods The stool samples of 150 patients (younger than 10 years) with diarrhea were collected in this cross-sectional study (November 2020 to December 2021). After cultivation of samples on Hektoen Enteric agar and xylose lysine deoxycholate agar, standard microbiology tests, VITEK 2 system, and polymerase chain reaction (PCR) were utilized to identify Shigella isolates. The broth microdilution method was used to determine antibiotic susceptibility. PMQR genes including qnrA, qnrB, qnrC, qnrD, qnrE, qnrS, qnrVC, qepA, oqxAB, aac(6′)-Ib-cr, and crpP and biofilm formation were investigated in quinolone-resistant isolates by PCR and microtiter plate method, respectively. An enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique was used to determine the clonal relatedness of quinolone-resistant isolates. Results A total of 95 Shigella isolates including S. sonnei (53, 55.8%), S. flexneri (39, 41.1%), and S. boydii (3, 3.2%) were identified. The highest resistance rates of the isolates were against ampicillin (92.6%, n = 88/95). Overall, 42 of 95 (44.2%) isolates were simultaneously resistant against two or more quinolones including 26 (61.9%) S. sonnei and 16 (38.1%) S. flexneri. All isolates were multidrug-resistant (resistance to more than 3 antibiotics). The occurrence of PMQR genes was as follows: qnrS (52.4%), qnrA and aac(6′)-Ib-cr (33.3%), and qnrB (19.0%). The prevalence in species was as follows: 61.5% and 37.5% (qnrS), 19.2% and 56.3% (qnrA), 38.5% and 25.0 (aac(6′)-Ib-cr), and 19.2% and 18.8% (qnrB) for S. sonnei and S. flexneri, respectively. The other PMQR genes were not detected. In total, 52.8% (28/53) of quinolone-susceptible and 64.3% (27/42) of quinolone-resistant isolates were biofilm producers. Biofilm formation was not significantly different between quinolone-resistant and quinolone-susceptible isolates (P-value = 0.299). Quinolone-resistant isolates showed a high genetic diversity according to the ERIC-PCR. Conclusion It seems that qnrS, qnrA, and aac(6′)-Ib-cr play a significant role in the quinolone resistance among Shigella isolates in our region. Also the quinolone-resistant S. flexneri and S. sonnei isolates had a high genetic diversity. Hence, antibiotic therapy needs to be routinely revised based on the surveillance findings.

Abstract Background The incidence of hospital-acquired infections in extensively drug-resistant Pseudomonas aeruginosa (XDR-PA) has been increasing worldwide and is frequently associated with an increase in mortality and morbidity rates. The aim of this study was to characterize clinical XDR-PA isolates recovered during six months at three different hospitals in Egypt. Results Seventy hospital-acquired clinical isolates of P. aeruginosa were classified into multidrug-resistant (MDR), extensively drug-resistant (XDR) and pandrug-resistant (PDR), according to their antimicrobial resistance profile. In addition, the possession of genes associated with mobile genetic elements and genes encoding antimicrobial resistance determinants among isolates were detected using polymerase chain reaction. As a result, a significant percentage of the isolates (75.7%) were XDR, while 18.5% were MDR, however only 5.7% of the isolates were non-MDR. The phenotypic detection of carbapenemases, extended-spectrum β-lactamases (ESBLs) and metallo β-lactamase (MBL) enzymes showed that 73.6% of XDR-PA isolates were carbapenemases producers, whereas 75.5% and 88.7% of XDR-PA isolates produced ESBLs and MBL respectively. In addition, PCR screening showed that oxa gene was the most frequently detected gene of carbapenemases (91.4%), while aac(6ʹ)-lb gene was mostly detected (84.3%) among the screened aminoglycosides-resistance genes. Furthermore, the molecular detection of the colistin resistance gene showed that 12.9% of isolates harbored mcr-1 gene. Concerning mobile genetic element markers (intI, traA, tnp513, and merA), intI was the highest detected gene as it was amplified in 67 isolates (95.7%). Finally, phylogenetic and molecular typing of the isolates via ERIC-PCR analysis revealed 10 different ERIC fingerprints. Conclusion The present study revealed a high prevalence of XDR-PA in hospital settings which were resistant to a variety of antibiotics due to several mechanisms. In addition, 98% of the XDR-PA clinical isolates contained at least one gene associated with movable genetic elements, which could have aided the evolution of these XDR-PA strains. To reduce spread of drug resistance, judicious use of antimicrobial agents and strict infection control measures are therefore essential.

Abstract Objectives In the recent years, multidrug resistant (MDR) neonatal septicemia-causing Enterobacterales has been dramatically increased due to the extended-spectrum beta-lactamases (ESBLs) and AmpC enzymes. This study aimed to assess the antibiotic resistance pattern, prevalence of ESBLs/AmpC beta-lactamase genes, and Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) fingerprints in Enterobacterales isolated from neonatal sepsis. Results In total, 59 Enterobacterales isolates including 41 (69.5%) Enterobacter species, 15 (25.4%) Klebsiella pneumoniae and 3 (5.1%) Escherichia coli were isolated respectively. Resistance to ceftazidime and cefotaxime was seen in all of isolates. Furthermore, all of them were multidrug-resistant (resistant to three different antibiotic categories). The phenotypic tests showed that 100% of isolates were ESBL-positive. Moreover, AmpC production was observed in 84.7% (n = 50/59) of isolates. Among 59 ESBL-positive isolates, the highest percentage belonged to bla CTX−M−15 gene (66.1%) followed by bla CTX−M (45.8%), bla CTX−M−14 (30.5%), bla SHV (28.8%), and bla TEM (13.6%). The frequency of bla DHA, bla EBC, bla MOX and bla CIT genes were 24%, 24%, 4%, and 2% respectively. ERIC-PCR analysis revealed that Enterobacterales isolates were genetically diverse. The remarkable prevalence of MDR Enterobacterales isolates carrying ESBL and AmpC beta-lactamase genes emphasizes that efficient surveillance measures are essential to avoid the more expansion of drug resistance amongst isolates.

Background: The incidence of hospital-acquired infections in extensively drug-resistant Pseudomonas aeruginosa (XDR-PA) has been increasing worldwide and is frequently associated with an increase in mortality and morbidity rates. The aim of this study was to characterize clinical XDR-PA isolates recovered during six months at three different hospitals in Egypt. Results: Seventy hospital-acquired clinical isolates of P. aeruginosa were classified into multidrug-resistant (MDR), extensively drug-resistant (XDR) and pandrug-resistant (PDR), according to their antimicrobial resistance profile. In addition, the possession of genes associated with mobile genetic elements and genes encoding antimicrobial resistance determinants among isolates were detected using polymerase chain reaction. As a result, a significant percentage of the isolates (75.7%) were XDR, while 18.5% were MDR, however only 5.7% of the isolates were non-MDR. The phenotypic detection of carbapenemases, extended-spectrum β-lactamases (ESBLs) and metallo β-lactamase (MBL) enzymes showed that 73.6% of XDR-PA isolates were carbapenemases producers, whereas 75.5% and 88.7% of XDR-PA isolates produced ESBLs and MBL respectively. In addition, PCR screening showed that oxa gene was the most frequently detected gene of carbapenemases (91.4%), while aac(6ʹ)-lb gene was mostly detected (84.3%) among the screened aminoglycosides-resistance genes. Furthermore, the molecular detection of the colistin resistance gene showed that 12.9% of isolates harbored mcr-1 gene. Concerning mobile genetic element markers (intI, traA, tnp513, and merA), intI was the highest detected gene as it was amplified in 67 isolates (95.7%). Finally, phylogenetic and molecular typing of the isolates via ERIC-PCR analysis revealed 10 different ERIC fingerprints. Conclusion: The present study revealed a high prevalence of XDR-PA in hospital settings which were resistant to a variety of antibiotics due to several mechanisms. In addition, 98% of the XDR-PA clinical isolates contained at least one gene associated with movable genetic elements, which could have aided the evolution of these XDR-PA strains. To reduce spread of drug resistance, judicious use of antimicrobial agents and strict infection control measures are therefore essential. [ABSTRACT FROM AUTHOR]

This study depicts the drug-resistance and phylogenomic characteristics of 365 Escherichia coli (EC) and 76 Klebsiella pneumoniae (KP) isolated from stray dogs (293) in and around Kolkata, India. Initial screening found 59 isolates, including 48 E. coli and 11 KP multidrug resistant, which included 33 extended-spectrum β-lactamase, 41 AmpC β-lactamase and 18 metallo-β-lactamase producers carrying blaNDM-1 (11) and blaNDM-5 (7) genes. Majority of them had the resistant genes such as blaCTX-M (33), blaTEM (18), blaSHV (4), blaOXA (17), blaFOX (2), blaDHA (2), blaCITM (15), blaCMY-2 (13), blaGES (2) and blaVEB (2), qnrS (15), qnrB (3), aac-6′-Ib-cr (14), tetA (26), tetB (14), sul-1 (25), armA (2) and rmtB (6), in addition to adherence genes such as csgA (33), fimA (27), fliC (13), sdiA (33), rcsA (38), and rpoS (39). They also carried plasmid of diverse replicon types of which IncFIA and FIB were the most frequent. Phylogrouping categorized most of the MDR E. coli in phylogroup A (20), B1 (14), and B2 (6). Enterobacteriaceae repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) showed genetic diversity of multidrug resistant isolates irrespective of their origin, resistance, and virulence types, differentiating the EC in five clades (A–E) and KP in four clades (A–D). As these stray dogs, which had no history or scope of previous antimicrobial therapy, were found to have contracted potential antimicrobial resistance pathogens, the role of environment in spread of such pathogens and further possibility of human infections cannot be ruled out. [ABSTRACT FROM AUTHOR]

Captive birds of prey are often used for pest control in urban areas, while also participating in falconry exhibitions. Traveling across the country, these birds may represent a public health concern as they can host pathogenic and zoonotic agents and share the same environment as humans and synanthropic species. In this work, Escherichia coli from the cloacal samples of 27 captive birds of prey were characterized to determine their pathogenic potential. Isolates were clustered through ERIC-PCR fingerprinting, and the phylogenetic groups were assessed using a quadruplex PCR method. Their virulence and resistance profile against nine antibiotics were determined, as well as the isolates' ability to produce extended-spectrum β-lactamases (ESBLs). The 84 original isolates were grouped into 33 clonal types, and it was observed that more than half of the studied isolates belonged to groups D and B2. Most isolates presented gelatinase activity (88%), almost half were able to produce biofilm (45%), and some were able to produce α-hemolysin (18%). The isolates presented high resistance rates towards piperacillin (42%), tetracycline (33%), and doxycycline (30%), and 6% of the isolates were able to produce ESBLs. The results confirm the importance of these birds as reservoirs of virulence and resistance determinants that can be disseminated between wildlife and humans, stressing the need for more studies focusing on these animals. [ABSTRACT FROM AUTHOR]

This study was conducted for identifying phylogenetic relationships between 15 scab-causing Streptomyces species including S. bottropensis, S. europaeiscabiei, S. scabiei, S. stelliscabiei and, other 11 Streptomyces sp. All of the strains were originally isolated from symptomatic potatoes in Erzurum Province, The Eastern Anatolia Region of Turkey. Some morphological and biochemical properties of the strains were defined in our former research. Then, 16 s rRNA regions of them were sequenced. After the sequence data assembly, phylogenetic analyzes were performed. The phylogenetic analyses revealed that the strains are involved in the same major group and, substantially similar to reference strains. Additionally, some subgroup formations were also recorded. Moreover, Repetitive element-based PCR (Rep-PCR), Enterobacterial repetitive intergenic consensus (ERIC-PCR), and BOX-PCR fingerprinting molecular typing methods were used for as molecular typing methods. According to our knowledge, this is the first report on phylogenetic relationships of scab-causing Streptomyces species from Turkey. However, the identification of most pathogenic strains remained at the species level. [ABSTRACT FROM AUTHOR]

Reem Aljindan,1 Reema Allahham,2 Rana Alghamdi,2 Ibrahim Alhabib,2 Samia AlNassri,3 Wala Alkhalifa,4 Asim Diab,1 Amer Alomar,2 Lamya Yamani,2 Nasreldin Elhadi2 1Department of Microbiology, College of Medicine, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom of Saudi Arabia; 2Department of Clinical Laboratory Science, College of Applied Medical Sciences, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom of Saudi Arabia; 3Department of Infection Control, King Fahad Hospital of the University, Alkhobar, Kingdom of Saudi Arabia; 4Department of Microbiology, Comprehensive Screening Center, Eastern Health Cluster, Dammam, Kingdom of Saudi ArabiaCorrespondence: Nasreldin Elhadi, Department of Clinical Laboratory Science, College of Applied Medical Sciences, Imam Abdulrahman Bin Faisal University, P.O. Box 2435, Dammam, 31441, Kingdom of Saudi Arabia, Email nmohammed@iau.edu.saBackground: Urinary tract infection (UTI) caused by V. cholerae is rare and less common. V. cholerae is a Gram-negative bacterium motile using single polar flagellum and, originally, is a waterborne microbe found in aquatic and estuarine environments. Toxigenic V. cholerae is well-known as a causative agent of acute and excessive watery diarrhea after ingesting food and water contaminated with this bacterium.Case Presentation: A 27-year-old male patient presented to the emergency department on 17th July 2021 with burning micturition, normal vital signs, and no fever, vomiting, or diarrhea. In 2017, the patient complained of short stature and vitamin D deficiency. He was on human growth hormone from January 2018 till October 2019. The diagnosis was V. cholerae Non-O1/non-O139 urinary tract infection (UTI). Considering a urinary tract infection, empirical treatment with Lornoxicam and Ciprofloxacin was initiated, while the result of urine culture was still pending. The patient was discharged on the same day and without any complications.Conclusion: V. cholerae non-O1/non-O139 is primarily a marine inhabitant and is associated with sporadic cases resulting in cholera-like diarrhea after consumption of contaminated seafood and exposure to seawater. Extraintestinal infection associated with this bacterium should no longer be ignored as this change in the behavior of cholera bacteria mechanism of pathogenicity might be related to some associated virulence genes.Keywords: non-O1/non-O139 V. cholerae, virulence genes, extraintestinal infection, ERIC-PCR

Abstract Background Water is considered a source for the transmission of Arcobacter species to both humans and animals. This study was conducted to assess the prevalence, distribution, and pathogenicity of A. butzleri strains, which can potentially pose health risks to humans and animals. Cultures were isolated from surface waters of a mixed-use but predominately agricultural watershed in eastern Ontario, Canada. The detection of antimicrobial resistance (AMR) and virulence-associated genes (VAGs), as well as enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) assays were performed on 913 A. butzleri strains isolated from 11 agricultural sampling sites. Results All strains were resistant to one or more antimicrobial agents, with a high rate of resistance to clindamycin (99%) and chloramphenicol (77%), followed by azithromycin (48%) and nalidixic acid (49%). However, isolates showed a significantly (p 85%) compared to irgA (25%), hecB (19%), hecA (15%), and cj1349 (12%) genes. Co-occurrence analysis showed A. butzleri strains resistant to clindamycin, chloramphenicol, nalidixic acid, and azithromycin were positive for ciaB, tlyA, mviN and pldA VAGs. ERIC-PCR fingerprint analysis revealed high genetic similarity among strains isolated from three sites, and the genotypes were significantly associated with AMR and VAGs results, which highlight their potential environmental ubiquity and potential as pathogenic. Conclusions The study results show that agricultural activities likely contribute to the contamination of A. butzleri in surface water. The findings underscore the importance of farm management practices in controlling the potential spread of A. butzleri and its associated health risks to humans and animals through contaminated water.

Mastitis is an inflammation of the mammary gland caused in dairy cows due to bacterial infections causing high economic losses. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) is an effective genotyping tool for tracing the infection by different bacteria. One hundred milk samples were collected (50 from clinical mastitis and 50 from subclinical mastitis) from different dairy farms at different regions of El-Gharbia governorate in Egypt. The samples were examined bacteriologically for the isolation and identification of Klebsiella pneumoniae and Corynebacterium bovis. Antibiotic sensitivity testing for the isolates and genotyping by ERIC-PCR were performed. Our results showed that the prevalence of K. pneumoniae was 41% from total samples and C. bovis strains was18% from subclinical mastitis milk samples. All the examined isolates were multi drug resistant with higher resistance to ampicillin, amoxicillin-clavulanate and cefotaxime for K. pneumoniae and to penicillin, erythromycin and tetracycline for C. bovis. Discriminatory index of ERIC-PCR was 0.984 and 1 for K. pneumoniae and C. bovis isolates, respectively. The dendrogram analysis for K. pneumoniae showed three clusters and two separate isolates, while for C. bovis 1 cluster with 2 sub clusters and three separate isolates were observed. It was concluded that ERIC-PCR is proven to be effective genotyping technique with high discriminatory index and is a good epidemiological tool for mastitis in cows as there was a genetic relatedness between some strains collected from different regions at El-Gharbia governorate in Egypt. This indicated the possibility of infection transmission between these regions and necessitates the need to increase control measures. [ABSTRACT FROM AUTHOR]

Background: Uropathogenic Escherichia coli (UPEC) is the main etiological agent behind community-acquired and hospital-acquired urinary tract infections (UTIs), which are among the most prevalent human infections. The management of UPEC infections is becoming increasingly difficult owing to multi-drug resistance, biofilm formation, and the possession of an extensive virulence arsenal. This study aims to characterize UPEC isolates in Tanta, Egypt, with regard to their antimicrobial resistance, phylogenetic profile, biofilm formation, and virulence, as well as the potential associations among these factors. Methods: One hundred UPEC isolates were obtained from UTI patients in Tanta, Egypt. Antimicrobial susceptibility was assessed using the Kirby-Bauer method. Extended-spectrum β-lactamases (ESBLs) production was screened using the double disk synergy test and confirmed with PCR. Biofilm formation was evaluated using the microtiter-plate assay and microscopy-based techniques. The phylogenetic groups of the isolates were determined. The hemolytic activity, motility, siderophore production, and serum resistance of the isolates were also evaluated. The clonal relatedness of the isolates was assessed using ERIC-PCR. Results: Isolates displayed elevated resistance to cephalosporins (90–43%), sulfamethoxazole-trimethoprim (63%), and ciprofloxacin (53%). Ninety percent of the isolates were multidrug-resistant (MDR)/ extensively drug-resistant (XDR) and 67% produced ESBLs. Notably, there was an inverse correlation between biofilm formation and antimicrobial resistance, and 31%, 29%, 32%, and 8% of the isolates were strong, moderate, weak, and non-biofilm producers, respectively. Beta-hemolysis, motility, siderophore production, and serum resistance were detected in 64%, 84%, 65%, and 11% of the isolates, respectively. Siderophore production was correlated to resistance to multiple antibiotics, while hemolysis was more prevalent in susceptible isolates and associated with stronger biofilms. Phylogroups B2 and D predominated, with lower resistance and stronger biofilms in group B2. ERIC-PCR revealed considerable diversity among the isolates. Conclusion: This research highlights the dissemination of resistance in UPEC in Tanta, Egypt. The evident correlation between biofilm and resistance suggests a resistance cost on bacterial cells; and that isolates with lower resistance may rely on biofilms to enhance their survival. This emphasizes the importance of considering biofilm formation ability during the treatment of UPEC infections to avoid therapeutic failure and/or infection recurrence. [ABSTRACT FROM AUTHOR]

Background: Water is considered a source for the transmission of Arcobacter species to both humans and animals. This study was conducted to assess the prevalence, distribution, and pathogenicity of A. butzleri strains, which can potentially pose health risks to humans and animals. Cultures were isolated from surface waters of a mixed-use but predominately agricultural watershed in eastern Ontario, Canada. The detection of antimicrobial resistance (AMR) and virulence-associated genes (VAGs), as well as enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) assays were performed on 913 A. butzleri strains isolated from 11 agricultural sampling sites. Results: All strains were resistant to one or more antimicrobial agents, with a high rate of resistance to clindamycin (99%) and chloramphenicol (77%), followed by azithromycin (48%) and nalidixic acid (49%). However, isolates showed a significantly (p < 0.05) high rate of susceptibility to tetracycline (1%), gentamycin (2%), ciprofloxacin (4%), and erythromycin (5%). Of the eight VAGs tested, ciaB, mviN, tlyA, and pldA were detected at high frequency (> 85%) compared to irgA (25%), hecB (19%), hecA (15%), and cj1349 (12%) genes. Co-occurrence analysis showed A. butzleri strains resistant to clindamycin, chloramphenicol, nalidixic acid, and azithromycin were positive for ciaB, tlyA, mviN and pldA VAGs. ERIC-PCR fingerprint analysis revealed high genetic similarity among strains isolated from three sites, and the genotypes were significantly associated with AMR and VAGs results, which highlight their potential environmental ubiquity and potential as pathogenic. Conclusions: The study results show that agricultural activities likely contribute to the contamination of A. butzleri in surface water. The findings underscore the importance of farm management practices in controlling the potential spread of A. butzleri and its associated health risks to humans and animals through contaminated water. Highlights: 1. Arcobacter butzleri was highly prevalent in agricultural surface water. 2. Strains were highly resistant to antimicrobial agents. 3. Virulence genes were detected at a high frequency. 4. High genetic similarity and association among strains from various sites. [ABSTRACT FROM AUTHOR]

Abstract Background Shigella spp., which are facultative anaerobic bacilli within the Enterobacteriaceae family, present a significant public health burden due to their role as prominent contributors to diarrheal diseases worldwide. A molecular analysis can facilitate the identification and assessment of outbreaks involving this bacterium. So, we aimed to investigate the antibiotic susceptibility pattern and clonal relatedness of clinical Shigella spp. isolates obtained from patients with diarrhea in Hormozgan province, South of Iran. Methods From 2019 to 2021, a cross-sectional investigation was conducted on 448 stool samples obtained from patients who were experiencing diarrhea, in the southern region of Iran. Shigella spp. isolates were identified based on biochemical and serological tests. All Shigella species were verified using species-specific polymerase chain reaction (PCR), followed by susceptibility testing to antimicrobial agents. Subsequently, genotyping of all Shigella species was conducted using ERIC-PCR. Results Out of a total of 448 stool samples, the presence of Shigella was detected in 62 cases, accounting for a prevalence rate of 13.84%. Among the identified isolates, the majority were attributed to S. flexneri, representing 53.23% of the cases. This was followed by S. sonnei at 24.19% and S. boydii at 22.58%. Notably, no instances of S. dysenteriae were found. The highest prevalence of Shigella isolates was observed in infants and children under the age of five. A significant proportion of the identified isolates demonstrated resistance to various antibiotics. Specifically, high resistance rates were noted for ampicillin (90.78%), piperacillin–tazobactam (87.1%), cefixime (83.87%), trimethoprim–sulfamethoxazole (83.87%), cefotaxime (82.26%), and ceftriaxone (80.65%). In addition, a substantial number (87.1%) of the isolates exhibited a multidrug-resistant (MDR) phenotype. Using the ERIC-PCR method, a total of 11 clusters and 6 distinct single types were identified among all the Shigella isolates. Conclusion A notable occurrence of antibiotic-resistant Shigella species has been noted, with multi-drug resistant (MDR) strains presenting an increasing challenge for treating shigellosis worldwide, and this includes Iran. Techniques such as ERIC-PCR are useful for assessing the genetic variation and connections between Shigella strains, which indirectly contributes to understanding antimicrobial resistance patterns. Further research is needed to explore the specific correlation between resistance genes and ERIC genotyping patterns in Shigella strains.

Background: Shigella spp., which are facultative anaerobic bacilli within the Enterobacteriaceae family, present a significant public health burden due to their role as prominent contributors to diarrheal diseases worldwide. A molecular analysis can facilitate the identification and assessment of outbreaks involving this bacterium. So, we aimed to investigate the antibiotic susceptibility pattern and clonal relatedness of clinical Shigella spp. isolates obtained from patients with diarrhea in Hormozgan province, South of Iran. Methods: From 2019 to 2021, a cross-sectional investigation was conducted on 448 stool samples obtained from patients who were experiencing diarrhea, in the southern region of Iran. Shigella spp. isolates were identified based on biochemical and serological tests. All Shigella species were verified using species-specific polymerase chain reaction (PCR), followed by susceptibility testing to antimicrobial agents. Subsequently, genotyping of all Shigella species was conducted using ERIC-PCR. Results: Out of a total of 448 stool samples, the presence of Shigella was detected in 62 cases, accounting for a prevalence rate of 13.84%. Among the identified isolates, the majority were attributed to S. flexneri, representing 53.23% of the cases. This was followed by S. sonnei at 24.19% and S. boydii at 22.58%. Notably, no instances of S. dysenteriae were found. The highest prevalence of Shigella isolates was observed in infants and children under the age of five. A significant proportion of the identified isolates demonstrated resistance to various antibiotics. Specifically, high resistance rates were noted for ampicillin (90.78%), piperacillin–tazobactam (87.1%), cefixime (83.87%), trimethoprim–sulfamethoxazole (83.87%), cefotaxime (82.26%), and ceftriaxone (80.65%). In addition, a substantial number (87.1%) of the isolates exhibited a multidrug-resistant (MDR) phenotype. Using the ERIC-PCR method, a total of 11 clusters and 6 distinct single types were identified among all the Shigella isolates. Conclusion: A notable occurrence of antibiotic-resistant Shigella species has been noted, with multi-drug resistant (MDR) strains presenting an increasing challenge for treating shigellosis worldwide, and this includes Iran. Techniques such as ERIC-PCR are useful for assessing the genetic variation and connections between Shigella strains, which indirectly contributes to understanding antimicrobial resistance patterns. Further research is needed to explore the specific correlation between resistance genes and ERIC genotyping patterns in Shigella strains. [ABSTRACT FROM AUTHOR]

The emergence of extended-spectrum β-lactamase-producing Klebsiella pneumoniae, including CRKP infections, has resulted in significant morbidity and mortality worldwide. We aimed to explore the presence of bla genes (CTX-M, TEM, and SHV) in CRKP isolates. A total of 24 CRKP isolates were randomly selected from the Salmaniya Medical Complex Microbiology Laboratory. These isolates, which were positive for carbapenemases, were further explored for CTX-M, TEM, and SHV genes using PCR. All the CTX-M PCR amplicons were sent for sequencing. To determine genetic relatedness, molecular typing by ERIC-PCR was performed. The bla gene testing demonstrated that a significant proportion of these isolates harbored SHV, CTX-M, and TEM genes (100%, 91.6%, and 45.8%), respectively. Bioinformatic analyses confirmed CTX-M-15 in these isolates. ERIC-PCR analysis showed three clusters demonstrating genetic relatedness. The study findings reveal the concomitant carriage of the SHV and CTX-M-15 and a comparatively lower carriage of TEM genes in CRKP isolates. Our findings highlight the significance of routinely reporting the presence of antibiotic resistance genes along with regular antibiotic sensitivity reports, as this will aid clinicians in prescribing appropriate antibiotics. [ABSTRACT FROM AUTHOR]

Klebsiella pneumoniae is a gram-negative bacterium that causes severe illnesses and is antibiotic-resistant. This study aimed to determine the antibiotic resistance profile and prevalence of classII and III Iintegrons and ERIC -PCR among clinical isolates of K. pneumoniae. The study was conducted between September 2022 and January 2023. Fifty isolates were obtained from 230 specimens (wounds, burns, blood, fluid, ears, urine and sputum). Macroscopic, microscopic, and biochemical assays were used to identify all K.pneumoniae isolates, which were confirmed with the genetically by 16S rRNA. All isolates were examined for various types of clinically significant antibiotic drugs. The results of resistance to antibiotics indicated resistance to Amoxillineclavulanc acid(98%), Meropenem (38%), Ceftazidime (96%), Amikacin (48%), Trimethopri-sulfamethoxazole (58.62%) and Levofloxacin (46%). The testing for antibiotic susceptibility of the K.pneumoniaeisolate showed that 24 (48%) of the isolates were multi-drug resistant (MDR). K.pneumoniae β-lactamase producers (ESBL) appeared33(66%). Enterobacterial repetitive intergenic consensus (ERIC) amplification of 16 clinical K. pneumoniae isolates showed 14 (87.5 %) of Each of them revealed at least one amplification band. ERIC-PCR typing found two groups, A and B, with identical antimicrobial resistance patterns within the same group. While IntegronII showed that 1(6.25 %) of K. pneumoniae isolates was integrase gene positive. Class III integrons were seen in all isolates at a rate of 16 (100%). Continuous monitoring and characterization of integrons and their associated gene cassettes could be helpful in controlling the rate of antibiotic resistance by planning to take preventive measures to hinder the spread of resistant strains. [ABSTRACT FROM AUTHOR]

Public health is at risk because Streptococcus agalactiae is increasingly linked to incidences of bovine mastitis in Egypt. In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) were used to explore the genotyping of several antibiotic resistant Streptococcus agalactiae isolated from bovine mastitis and retail milk. Also, antimicrobial resistance genes were detected. Two hundred and sixty-six (46.7%) strains were isolated from samples of milk obtained from cases of bovine mastitis in dairy farms and retail markets representing 34 (5.96%) Streptococcus agalactiae and 232 (40.7%) other Streptococcus species strains based bacterial identification. By using disc diffusion assays, it was examined the susceptibility of all Streptococcus isolates to twelve antimicrobial agents. The highest prevalence of resistance of Streptococcus species was observed against ampicillin (65.5%), amoxicillin (56%), tetracycline (52.5%), ofloxacin (47.8%), and nitrofurantoin (46.9%). High proportion of the S. agalactiae isolates were resistance to amoxicillin (83%), followed by tetracycline (82.4%), nitrofurantoin (64.7%), azithromycin (61.8%), and ampicillin (50%). Most (88.2%) of S. agalactiae showed multiple antibiotic resistances (MAR) phenotypes with MAR index of 0.1-0.7 and 28 different MAR patterns. The results of genetic antimicrobial resistance of S. agalactiae strains revealed amplification of blaTEM (23%) and blaCTX (26%) genes in β-lactam-resistant strains, erm(B) gene (20%), mef(A) (35%) in macrolides resistant isolates, and tet(M) in (44%) tetracycline resistant strains. Using ERIC-PCR, the present study showed the genetic diversity and heterogeneity among S. agalactiae strains (n=34) that were classified into 19 distinct ERIC-PCR groups (A-S). Between them, ERIC O (20.6%, 7) was the most widespread group. These results indicated that milk samples served as source of MAR S. agalactiae, consequently posing threats to public health, so the improvement of the hygiene regimen on the farms and promotion of the wise use of antimicrobials are necessary. The obtained findings showed that milk samples were a source of MAR S. agalactiae, endangering public health. As a result, it is essential to enhance farm hygiene practices and promote the responsible use of antimicrobials. Additional research on the epidemiology of S. agalactiae is required to add bacterial genetic information in order to help in rational vaccine strategy in the future. [ABSTRACT FROM AUTHOR]

Aims: Evaluate methods for identification and typing of Stenotrophomonas maltophilia isolated from a pharmaceutical facility. Methods and results: From 270 S. maltophilia strains identified by VITEK®2, 40 were selected and submitted to MALDI TOF-MS, 16S and 23S rRNA gene analysis, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), and an antimicrobial susceptibility profile. 16S rRNA sequencing was able to identify 39 (97.5%) strains as Stenotrophomonas spp. and one (2.5%) as Luteimonas huabeiensis. MALDI TOF-MS identified 37 (92.5%) strains as S. maltophilia, and three (7.5%) were not identified. PCR targeting 23S rRNA yielded a positive result for 39 (97.5%) strains. However, after sequencing, two strains were identified as Stenotrophomonas rhizophila, showing false-positive results. The confirmed S. maltophilia strains (n = 37) showed 35 distinct ERIC-PCR profiles and exhibited sensitivity to minocycline and levofloxacin, and six (16.3%) showed intermediate resistance to sulfamethoxazole-trimethoprim. Conclusion: Matrix-assisted laser desorption lonization-time of flight mass spectrometry (MALDI-TOF MS) was a satisfactory methodology for the identification of S. maltophilia, but expansion of the database is necessary for the identification of other species. 16S rDNA sequencing showed low resolution for Stenotrophomonas species differentiation. PCR targeting 23S rRNA could not differentiate S. maltophilia from S. rhizophila. ERIC-PCR was shown to be a useful tool for the microbial source tracking of S. maltophilia. [ABSTRACT FROM AUTHOR]

Background. Enterococcus faecium is an opportunistic pathogen of humans with diverse hosts, encompassing animals as well as human beings. In the past twenty years, there has been a rise in the instances of nosocomial infections that are linked to antibiotic-resistant Enterococcus faecium. The acquisition of diverse antimicrobial resistance factors has driven the global development of robust and convergent adaptive mechanisms within the healthcare environment. The presence of microorganisms in hospitalized and non-hospitalized patient populations has been significantly aided by the facilitation of various perturbations within their respective microbiomes. Objective. This study aimed to determine the antimicrobial profile, demographic and clinical characteristics, along with the detection of virulence encoding genes, and to find out the clonal genetic relationship among colonized E. faecium strains. Methodology. A hospital-based cross-sectional study was carried out between October 2018 and March 2020 at four Khartoum locality hospitals in Sudan. The study comprised a total of 108 strains of E. faecium isolated from patients admitted to four locality hospitals in Khartoum. A self-structured questionnaire was used to gather information on sociodemographic traits. Data were analyzed using chi-square test. In all cases, P value ≤ 0.05 with a corresponding 95% confidence interval was considered statistically significant. Moreover, enterobacterial repetitive intergenic consensus--polymerase chain reaction (ERIC-PCR) was utilized to assess the prevalence of clonal relationships, and the gel was analyzed using CLIQS software. Results. In this study, the isolation rate of colonized E. faecium strains was 108/170 (63.5%). The colonization of E. faecium and its association with various sociodemo-graphic and clinical features was examined. 73 (67.6%) of patients had multidrug-resistant (MDR), and 22 (20.4%) had extensively drug-resistant (XDR), 73 (67.6%) of patients engaged in self-medication practices. Eighty patients (74.1%) were non-adherence to prescribed antibiotics, while 70 (64.8%) patients reported recent antibiotic usage within the 3 months. The present study suggests that demographic factors may not be significantly associated with the incidence of E. faecium infection except for patients who had a prior history of antibiotic use (P ≤ 0.005). The analysis of virulence genes showed a high prevalence of asa1 gene (22.2%) among strains. In ERIC-PCR the genetic relatedness of E. faecium showed seven identical clusters (A-G) with 100% genetic similarity. This implies clonal propagation in hospitals and communities. Conclusion. This study found that the incidence of E. faecium isolated from locality hospitals in Khartoum was likely due to the spread of E. faecium clones, thereby highlighting the need for intensifying infection control measures to prevent the spreading of nosocomial infection. [ABSTRACT FROM AUTHOR]

Mastitis is an inflammation of the mammary gland caused in dairy cows due to bacterial infections causing high economic losses. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) is an effective genotyping tool for tracing the infection by different bacteria. One hundred milk samples were collected (50 from clinical mastitis and 50 from subclinical mastitis) from different dairy farms at different regions of El-Gharbia Governorate in Egypt. The samples were examined bacteriologically for the isolation and identification of Klebsiella pneumoniae and Corynebacterium bovis. Antibiotic sensitivity testing for the isolates and genotyping by ERIC-PCR were performed. Our results showed that the prevalence of K. pneumoniae was 41% from total samples and C. bovis strains was18% from subclinical mastitis milk samples. All the examined isolates were multi drug resistant with higher resistance to ampicillin, amoxicillin-clavulanate and cefotaxime for K. pneumoniae and to penicillin, erythromycin and tetracycline for C. bovis. Discriminatory index of ERIC-PCR was 0.984 and 1 for K. pneumoniae and C. bovis isolates, respectively. The dendrogram analysis for K. pneumoniae showed three clusters and two separate isolates, while for C. bovis 1 cluster with 2 sub clusters and three separate isolates were observed. It was concluded that ERIC-PCR is proven to be effective genotyping technique with high discriminatory index and is a good epidemiological tool for mastitis in cows as there was a genetic relatedness between strains collected from different regions at El-Gharbia Government in Egypt. This indicated the possibility of infection transmission between these regions and necessitates the need to increase control measures.

ABSTRACT: Salmonella is a significant foodborne pathogen that has a significant impact on public health, and different strains of multidrug resistance (MDR) have been identified in this genus. This study used a combination of phenotypic and genotypic approaches to identify distinct Salmonella species collected from poultry broiler and layer farms, and antibiotic sensitivity testing was performed on these species. A total of 56 Salmonella isolates were serotyped, and phenotypic antibiotic resistance was determined for each strain. The enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) method was also used to provide a genotypic description, from which a dendrogram was constructed and the most likely phylogenetic relationships were applied. Salmonella isolates were detected in 20 (17%) out of 117 samples collected from small-scale broiler flocks. Salmonella isolates were classified as MDR strains after showing tolerance to 4 antibiotics, but no resistance to cloxacillin, streptomycin, vancomycin, or netilmicin was observed. From a genotypic perspective, these strains lack dfrD, parC, and blasfo-1 resistant genes, while harboring blactx-M, blaDHA-L, qnrA, qnrB, qnrS, gyrA, ermA, ermB, ermC, ermTR, mefA, msrA, tet A, tet B, tet L, tet M resistance genes. The genotyping results obtained with ERIC-PCR allowed isolates to be classified based on the source of recovery. It was determined that Salmonella strains displayed MDR, and many genes associated with them. Additionally, the ERIC-PCR procedure aided in the generation of clusters with biological significance. Extensive research on Salmonella serotypes is warranted, along with the implementation of long-term surveillance programs to monitor MDR Salmonella serotypes in avian-derived foods.

Abstract Ready‐to‐eat (RTE) foods are widely marketed in China and are important components of everyday diet. In this study, a total of 2000 RTE food samples were analyzed, 252 (12.60%) of which were positive for Enterobacteriaceae, and 48 were identified as containing extended‐spectrum β‐lactamase (ESBL)‐producing Escherichia coli isolates. Furthermore, the antimicrobial resistance patterns of these isolates to 14 antimicrobial agents revealed that most isolates were resistant to aminoglycosides and β‐lactam antibiotics. The TEM‐type gene was prevalent in our isolates (79.17%). The isolates (n = 48) were classified into three clusters based on the ERIC‐PCR results. Forty‐eight sequence types were found without duplicates, revealing genetic variation and relatedness among isolates. Thus, the results demonstrated the presence of ESBL‐producing Enterobacteriaceae in Chinese RTE foods. The results of this study provide insights into the spread of antibiotic‐resistant strains and improve understanding of microbial risks.

Background: Klebsiella pneumoniae (K. pneumoniae) is a significant contributor to nosocomial infections in neonates and children. This is due to a variety of virulence mechanisms, such as the ability to form biofilms and antibiotic resistance. Hence, this study aimed to evaluate the prevalence of K. pneumoniae infections among children, to determine the association of their antimicrobial resistance patterns, biofilm formation ability, and their molecular genotypes. Methods: Forty-six K. pneumoniae isolates were collected from a pediatric hospital in Alexandria, Egypt. After being identified by conventional methods, they were tested for their susceptibility to different classes of antibiotics, biofilm formation ability, and were genotyped by enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR). Results: Klebsiella pneumoniae was the most prevalent bacteria isolated from children (36%). Antimicrobial susceptibility testing revealed that the majority of our isolates (47.8%) were extensively drug-resistant (XDR) and 41.3 % were multi-drug resistant (MDR). Among the isolates, 80.4% were biofilm producers. No statistically significant association was noticed between the biofilm-producing ability and the drug-resistance type. Based on ERIC-PCR profile results, isolates were classified into 3 main clusters and 30 different ERIC genotypes. Moreover, no statistically significant association was found between ERIC-PCR clusters and neither the ability to produce biofilm nor the drug-resistance type. Conclusions: The results of this study indicate an alarming increase in the antimicrobial resistance patterns among K. pneumoniae isolated from neonates and children. ERIC-PCR typing showed high genetic diversity among K. pneumoniae isolates, indicating a polyclonal distribution of K. pneumoniae isolates within the hospital averments.

The goal of this study is to discover the genetic diversity of twelve Escherichia coli isolates harbor virulence genes (Stx1 and Stx2) isolated from farm fish and local market fish in different locations of Nineveh Governorate using Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) technique in the time span November, 2021 - January, 2022. The source of contamination was found depending on the fingerprint analysis of the repetitive polymerase chain reaction among the genes of the intestinal micro-organism ERIC-PCR for every isolate. The results show that there are similarities among the isolates and it ranged about 51%-100%. The isolates are divided into 7 genotypes, based primarily on a 90% similarity threshold 1 to 7, with the most offspring being inside genotype 1, since genotype 1 used to be the biggest team containing four isolates. Then, two isolates are grouped under genotypes 3 and 5. In contrast, genotype 2, 4, 6 and 7 have only one isolate. By virtue of genetic variations in the location, there was once an excessive clonal dispersion of most E. coli strains. In addition, the study shows genetic differences in E. coli in 83.3%, (10/12) of the isolates, in the different geographical locations of Mosul city. The E. coli isolates were classified into some groups: first, different genetic grouping, different geographical locations, same genotype; second, different genetic grouping, same geographical locations, different genotypes.

Captive birds of prey are often used for pest control in urban areas, while also participating in falconry exhibitions. Traveling across the country, these birds may represent a public health concern as they can host pathogenic and zoonotic agents and share the same environment as humans and synanthropic species. In this work, Escherichia coli from the cloacal samples of 27 captive birds of prey were characterized to determine their pathogenic potential. Isolates were clustered through ERIC-PCR fingerprinting, and the phylogenetic groups were assessed using a quadruplex PCR method. Their virulence and resistance profile against nine antibiotics were determined, as well as the isolates’ ability to produce extended-spectrum β-lactamases (ESBLs). The 84 original isolates were grouped into 33 clonal types, and it was observed that more than half of the studied isolates belonged to groups D and B2. Most isolates presented gelatinase activity (88%), almost half were able to produce biofilm (45%), and some were able to produce α-hemolysin (18%). The isolates presented high resistance rates towards piperacillin (42%), tetracycline (33%), and doxycycline (30%), and 6% of the isolates were able to produce ESBLs. The results confirm the importance of these birds as reservoirs of virulence and resistance determinants that can be disseminated between wildlife and humans, stressing the need for more studies focusing on these animals.

Objectives: This study aimed to assess the antimicrobial resistance profile, virulence potential, and genetic characterization of avian pathogenic Escherichia coli (APEC) that cause colibacillosis in poultry. Methods: Antibiotic susceptibility testing (AST) was measured via the Kirby-Bauer disc diffusion method against 27 commonly used antibiotics. Phylogrouping, virulence-associated gene detection, and hybrid strain detection via multiplex polymerase chain reaction (PCR) and genetic diversity were analysed via ERIC-PCR fingertyping method. Results: AST analysis showed 100% of isolates were multidrug-resistant (MDR) and highest resistance was against penicillin, tetracycline, and macrolide classes of antibiotics. The mcr-1 gene was present in 40% of the isolates, though only 4% of isolates were showing phenotypic resistance. Despite the scarce use of fluoroquinolone, carbapenem, and cephalosporin in the poultry sector, resistance was evident because of the high prevalence of extended-spectrum β-lactamase (ESBL) (53.7%) and other β-lactamases in APEC isolates. β-lactamase genotyping of APEC isolates revealed that 85.7% of isolates contained either blaCTX or blaTEM and around 38% of isolates were complement resistant. Growth in human urine was evident in 67.3% of isolates. Phylogroup B1 (51%) was the most prevalent group followed by phylogroups A (30.6%), D (13.61%), and B2 (4.76%). The most prevalent virulence-associated genes were fimH, iss, and tatT. Results showed that 26 isolates (17.69%) can be termed hybrid strains and APEC/EHEC (enterohemorrhagic E. coli) was the most prevalent hybrid E. coli pathotype. ERIC-PCR fingerprinting genotype analysis clustered APEC isolates in 40 groups (E1-E40). This study provides insights into the antibiotic resistance and virulence profiling of the APEC isolates in Pakistan. Conclusions: The findings of this study provide insights into that the antibiotic resistance and virulence profiling of the APEC isolates in Pakistan. This data can inform future studies designed to better estimate the severity of the colibacillosis in poultry farms.

Background: Caseous lymphadenitis (CLA) is a chronic suppurative bacterial infection caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) affecting superficial and internal lymph nodes and internal organs of small ruminants. Aims: Through the use of molecular methods, this study aimed to estimate the prevalence of caseous lymphadenitis and it's contributing factors as well as the degree of genetic diversity and epidemiological relationships among C. pseudotuberculosis isolates from slaughtered sheep and goats in various districts of Duhok Province, Iraq. Methods: A total of 18836 carcasses (15775 sheep and 3061 goats) were inspected by veterinarians at slaughterhouses (Duhok-Sumel (2453 sheep+627 goats), Zakho (6000 sheep), Bardarash (1117 sheep+379 goats), Amedi (413 sheep+178 goats) and Akre (5792 sheep+1877 goats)) in Duhok Province for detection the prevalence rate of CLA using molecular techniques. Results: The prevalence of the disease was 0.94% and 1.93% in sheep and goats, respectively. Sheep in Duhok-Sumel and goats in Amedi were at the higher risk of infection than the animals in another location, with a prevalence rate of 4.31% and 6.18%, respectively. Sheep and goats of older age were more susceptible. Females were more susceptible than males in all districts except Duhok-Sumel where the reverse was true. ERIC-PCR analysis grouped the bacterial isolates into 11 different genotypes. Maximum Likelihood phylogenetic tree of partial sequences of the 16S rRNA gene sequences of C. pseudotuberculosis revealed no divergent sequences discovered in this study. Conclusion: A strict control program needs to be applied to reduce the entrance of the pathogen from the neighboring countries. [Open Vet J 2023; 13(5.000): 588-598]

Background: Caseous lymphadenitis (CLA) is a chronic suppurative bacterial infection caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) affecting superficial and internal lymph nodes and internal organs of small ruminants. Aims: Through the use of molecular methods, this study aimed to estimate the prevalence of CLA and its contributing factors as well as the degree of genetic diversity and epidemiological relationships among C. pseudotuberculosis isolates from slaughtered sheep and goats in various districts of Duhok Province, Iraq. Methods: A total of 18,836 carcasses (15,775 sheep and 3,061 goats) were inspected by veterinarians at slaughterhouses [Duhok-Sumel (2,453 sheep + 627 goats), Zakho (6,000 sheep), Bardarash (1,117 sheep + 379 goats), Amedi (413 sheep + 178 goats) and Akre (5,792 sheep + 1,877 goats)] in Duhok Province for detection the prevalence rate of CLA using molecular techniques. Results: The prevalence of the disease was 0.94% and 1.93% in sheep and goats, respectively. Sheep in Duhok-Sumel and goats in Amedi were at a higher risk of infection than the animals in another location, with a prevalence rate of 4.31% and 6.18%, respectively. Sheep and goats of older age were more susceptible. Females were more susceptible than males in all districts except Duhok-Sumel where the reverse was true. ERIC-PCR analysis grouped the bacterial isolates into 11 different genotypes. The maximum likelihood phylogenetic tree of partial sequences of the 16S rRNA gene sequences of C. pseudotuberculosis revealed no divergent sequences discovered in this study. Conclusion: A strict control program needs to be applied to reduce the entrance of pathogen from neighboring countries. [ABSTRACT FROM AUTHOR]

Ready‐to‐eat (RTE) foods are widely marketed in China and are important components of everyday diet. In this study, a total of 2000 RTE food samples were analyzed, 252 (12.60%) of which were positive for Enterobacteriaceae, and 48 were identified as containing extended‐spectrum β‐lactamase (ESBL)‐producing Escherichia coli isolates. Furthermore, the antimicrobial resistance patterns of these isolates to 14 antimicrobial agents revealed that most isolates were resistant to aminoglycosides and β‐lactam antibiotics. The TEM‐type gene was prevalent in our isolates (79.17%). The isolates (n = 48) were classified into three clusters based on the ERIC‐PCR results. Forty‐eight sequence types were found without duplicates, revealing genetic variation and relatedness among isolates. Thus, the results demonstrated the presence of ESBL‐producing Enterobacteriaceae in Chinese RTE foods. The results of this study provide insights into the spread of antibiotic‐resistant strains and improve understanding of microbial risks. [ABSTRACT FROM AUTHOR]

Streptococcus pneumoniae serotype 19A is frequently isolated worldwide. In this study, the clonal relationships among 62 isolates from different patients from 21 hospitals between 2008 and 2018 were characterized using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The different band patterns that appeared upon agarose gel electrophoresis were used to construct an unweighted pair group method with an arithmetic mean (UPGMA) dendrogram. There were 23 different ERIC types (E1–E23). The most prevalent type was E9, accounting for 19.36% of all isolates, followed by E6 at 16.13%, E5 at 11.30%, and 13 ERIC types present in only one isolate or at 1.61% each. Using an additional study to determine the clonal relationships, we compared our ERIC-PCR results to the corresponding multilocus sequence types (MLSTs) from our recent study with the same 62 S. pneumoniae serotype 19A isolates. The results showed there were 20 different MLST types and that ERIC-PCR was comparable to MLST as a valuable complementary tool for the investigation of S. pneumoniae serotype 19A isolates. Furthermore, ERIC-PCR is very fast, affordable, and easy to perform compared to MLST. However, there is less concordance between these two methods. These results suggest a high diversity of different ERIC-PCR and MLST patterns. Overall, the combination of results from both methods can add greater discrimination and complementary information for the differentiation of S. pneumoniae strains in Thailand. [ABSTRACT FROM AUTHOR]

Background: Klebsiella pneumoniae (K. pneumoniae) is a significant contributor to nosocomial infections in neonates and children. This is due to a variety of virulence mechanisms, such as the ability to form biofilms and antibiotic resistance. Hence, this study aimed to evaluate the prevalence of K. pneumoniae infections among children, to determine the association of their antimicrobial resistance patterns, biofilm formation ability, and their molecular genotypes. Methods: Forty-six K. pneumoniae isolates were collected from a pediatric hospital in Alexandria, Egypt. After being identified by conventional methods, they were tested for their susceptibility to different classes of antibiotics, biofilm formation ability, and were genotyped by enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR). Results: Klebsiella pneumoniae was the most prevalent bacteria isolated from children (36%). Antimicrobial susceptibility testing revealed that the majority of our isolates (47.8%) were extensively drug-resistant (XDR) and 41.3 % were multi-drug resistant (MDR). Among the isolates, 80.4% were biofilm producers. No statistically significant association was noticed between the biofilm-producing ability and the drug-resistance type. Based on ERIC-PCR profile results, isolates were classified into 3 main clusters and 30 different ERIC genotypes. Moreover, no statistically significant association was found between ERICPCR clusters and neither the ability to produce biofilm nor the drugresistance type. Conclusions: The results of this study indicate an alarming increase in the antimicrobial resistance patterns among K. pneumoniae isolated from neonates and children. ERIC-PCR typing showed high genetic diversity among K. pneumoniae isolates, indicating a polyclonal distribution of K. pneumoniae isolates within the hospital averments. [ABSTRACT FROM AUTHOR]

Background Enterococcus faecium is an opportunistic pathogen of humans with diverse hosts, encompassing animals as well as human beings. In the past twenty years, there has been a rise in the instances of nosocomial infections that are linked to antibiotic-resistant Enterococcus faecium. The acquisition of diverse antimicrobial resistance factors has driven the global development of robust and convergent adaptive mechanisms within the healthcare environment. The presence of microorganisms in hospitalized and non-hospitalized patient populations has been significantly aided by the facilitation of various perturbations within their respective microbiomes. Objective This study aimed to determine the antimicrobial profile, demographic and clinical characteristics, along with the detection of virulence encoding genes, and to find out the clonal genetic relationship among colonized E. faecium strains. Methodology A hospital-based cross-sectional study was carried out between October 2018 and March 2020 at four Khartoum locality hospitals in Sudan. The study comprised a total of 108 strains of E. faecium isolated from patients admitted to four locality hospitals in Khartoum. A self-structured questionnaire was used to gather information on sociodemographic traits. Data were analyzed using chi-square test. In all cases, P value ≤ 0.05 with a corresponding 95% confidence interval was considered statistically significant. Moreover, enterobacterial repetitive intergenic consensus–polymerase chain reaction (ERIC-PCR) was utilized to assess the prevalence of clonal relationships, and the gel was analyzed using CLIQS software. Results In this study, the isolation rate of colonized E. faecium strains was 108/170 (63.5%). The colonization of E. faecium and its association with various sociodemographic and clinical features was examined. 73 (67.6%) of patients had multidrug-resistant (MDR), and 22 (20.4%) had extensively drug-resistant (XDR), 73 (67.6%) of patients engaged in self-medication practices. Eighty patients (74.1%) were non-adherence to prescribed antibiotics, while 70 (64.8%) patients reported recent antibiotic usage within the 3 months. The present study suggests that demographic factors may not be significantly associated with the incidence of E. faecium infection except for patients who had a prior history of antibiotic use (P ≤ 0.005). The analysis of virulence genes showed a high prevalence of asa1 gene (22.2%) among strains. In ERIC-PCR the genetic relatedness of E. faecium showed seven identical clusters (A–G) with 100% genetic similarity. This implies clonal propagation in hospitals and communities. Conclusion This study found that the incidence of E. faecium isolated from locality hospitals in Khartoum was likely due to the spread of E. faecium clones, thereby highlighting the need for intensifying infection control measures to prevent the spreading of nosocomial infection.

Simple Summary: The present study aimed to isolate Corynebacterium pseudotuberculosis from caseous lymphadenitis (CLA) in smallholder sheep and goats. Thereafter, antimicrobial resistance determinants, virulence characteristics, and phylogenetic analysis of the isolates were investigated. Our results revealed that 24.54% (54/220) of examined animals showed CLA-compatible lesions from which C. pseudotuberculosis was isolated. Antimicrobial susceptibility testing revealed that all isolates were resistant to bacitracin and florfenicol. Additionally, 92.6% of the isolates displayed resistance to penicillin and erythromycin, while none of the isolates were resistant to norfloxacin. Interestingly, 16.7% of C. pseudotuberculosis isolates recovered from sheep showed vancomycin resistance. Molecular characterization revealed that PLD, PIP, and FagA virulence genes were detected in all examined C. pseudotuberculosis isolates. The bla (β-lactam) resistance gene was present in all isolates. Additionally, 83% of sheep isolates carried the aminoglycoside (aph(3″)-lb), chloramphenicol (cat1), and bacitracin (bcrA) resistance genes. Notably, the glycopeptide (vanA) resistance gene was detected in 8% of the sheep isolates as a first report. Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) genotyping of 10 multi-drug resistant C. pseudotuberculosis isolates showed a high similarity index of 83.6% between isolates from sheep and goats. Nucleotide sequence analysis of partial 16S rRNA sequences of these isolates showed 98.83% similarities with biovar Ovis of globally available reference sequences on the Genbank database. In conclusion, this study emphasized the high isolation rate of C. pseudotuberculosis and its molecular characterization in smallholder sheep and goats in four districts in Egypt. Caseous lymphadenitis (CLA) is a bacterial infection caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) that affects sheep and goats, leading to abscess formation in their lymph nodes. The present study aimed to isolate and identify C. pseudotuberculosis from CLA in smallholder sheep and goats, and determine the resistance patterns, virulence, and resistance genes of the isolates. Additionally, genotypic and phylogenetic analysis of the isolates was conducted using ERIC-PCR and DNA sequencing techniques. A cross-sectional study examined 220 animals (130 sheep and 90 goats) from 39 smallholder flocks for clinical signs of CLA. Fifty-four (24.54%) animals showed CLA-compatible lesions, confirmed by C. pseudotuberculosis isolation and PCR identification. Sheep had a lower infection rate of CLA (18.46%) compared with goats (33.3%). Antimicrobial susceptibility testing of 54 C. pseudotuberculosis isolates to 24 antimicrobial drugs revealed that they were 100% resistant to bacitracin and florfenicol, while none of the isolates were resistant to norfloxacin. A high resistance rate was observed for penicillin and erythromycin (92.6% each). Interestingly, 16.7% of C. pseudotuberculosis isolates recovered from sheep showed vancomycin resistance. Molecular characterization of C. pseudotuberculosis isolates revealed that PLD, PIP, and FagA virulence genes were present in all examined isolates. However, the FagB, FagC, and FagD genes were detected in 24 (100%), 20 (83%), and 18 (75%) of the sheep isolates, and 26 (87%), 26 (87%), and 18 (60%) of the goat isolates, respectively. The β-lactam resistance gene was present in all isolates. Furthermore, 83% of the sheep isolates carried the aminoglycoside (aph(3″)-lb), chloramphenicol (cat1), and bacitracin (bcrA) resistance genes. Among the isolates recovered from goats, 73% were found to contain macrolides (ermX), sulfonamide (sul1), and bacitracin (bcrA) resistance genes. It is worrisome that the glycopeptide (vanA) resistance gene was detected in 8% of the sheep isolates as a first report. ERIC-PCR genotyping of 10 multi-drug-resistant C. pseudotuberculosis isolates showed a high similarity index of 83.6% between isolates from sheep and goats. Nucleotide sequence analysis of partial 16S rRNA sequences of C. pseudotuberculosis revealed 98.83% similarity with biovar Ovis of globally available reference sequences on the Genbank database. Overall, our findings might indicate that C. pseudotuberculosis infection in smallholders in Egypt might be underestimated despite the significant financial impact on animal husbandry and potential health hazards it poses. Moreover, this study highlights the importance of implementing a sustainable control strategy and increasing knowledge and awareness among smallholder breeders to mitigate the economic impact of CLA. [ABSTRACT FROM AUTHOR]

• ESKAPE bacteria was related in COVID-19/VAP patients. • MDR and XDR ESKAPE bacteria were identified. • Clonal dispersion of A. baumannii was detected in COVID-19/VAP patients. A decrease of detection of outbreaks by multidrug-resistant bacteria in critical areas has been reduced due to COVID-19 pandemic. Therefore, molecular epidemiological surveillance should be a primary tool to reveal associations not evident by classical epidemiology. The aim of this work was to demonstrate the presence of hidden outbreaks in the first wave of the COVID-19 pandemic and to associate their possible origin. A population of 96 COVID-19 patients was included in the study (April to June 2020) from Hospital Juárez de México. Genetic identification and antimicrobial susceptibility testing of VAP causative agents isolated from COVID-19 patients was performed. Resistance phenotypes were confirmed by PCR. Clonal association of isolates was performed by analysis of intergenic regions obtained. Finally, the association of clonal cases of VAP patients was performed by timelines. ESKAPE and non-ESKAPE bacteria were identified as causative agents of VAP. ESKAPE bacteria were classified as MDR and XDR. Only A. baumannii and P. aeruginosa were identified as clonally distributed in 13 COVID-19/VAP patients. Time analysis showed that cross-transmission existed between patients and care areas. Acinetobacter baumannii and Pseudomonas aeruginosa were involved in outbreaks non-detected in COVID-19/VAP patients in the first wave of COVID-19 pandemic. [ABSTRACT FROM AUTHOR]

Copyright of Harran University Journal of the Faculty of Veterinary Medicine / Harran Üniversitesi Veteriner Fakültesi Dergisi is the property of Harran University, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The threat of bacterial septicemia caused by Aeromonas hydrophila infection to aquaculture growth can be prevented through vaccination, but differences among A. hydrophila strains may affect the effectiveness of non-conserved subunit vaccines or non-inactivated A. hydrophila vaccines, making the identification and development of conserved antigens crucial. In this study, a bioinformatics analysis of 4268 protein sequences encoded by the A. hydrophila J-1 strain whole genome was performed based on reverse vaccinology. The specific analysis included signal peptide prediction, transmembrane helical structure prediction, subcellular localization prediction, and antigenicity and adhesion evaluation, as well as interspecific and intraspecific homology comparison, thereby screening the 39 conserved proteins as candidate antigens for A. hydrophila vaccine. The 9 isolated A. hydrophila strains from diseased fish were categorized into 6 different molecular subtypes via enterobacterial repetitive intergenic consensus (ERIC)-PCR technology, and the coding regions of 39 identified candidate proteins were amplified via PCR and sequenced to verify their conservation in different subtypes of A. hydrophila and other Aeromonas species. In this way, conserved proteins were screened out according to the comparison results. Briefly, 16 proteins were highly conserved in different A. hydrophila subtypes, of which 2 proteins were highly conserved in Aeromonas species, which could be selected as candidate antigens for vaccines development, including type IV pilus secretin PilQ (AJE35401.1) and TolC family outer membrane protein (AJE35877.1). The present study screened the conserved antigens of A. hydrophila by using reverse vaccinology, which provided basic foundations for developing broad-spectrum protective vaccines of A. hydrophila. [ABSTRACT FROM AUTHOR]

Urmia Lake is the largest lake in the Iranian plateau and the second largest Salt Lake in the world. This study was conducted to identify hypersaline bacteria in the lake through the screening with molecular markers. For the molecular study of the bacterial microbiome of the lake, samples were collected from water, sludge and soil of the different parts of the lake during different seasons of 2018 and 2019, and then transferred to the laboratory under standard conditions. Bacterial isolates were purified from the samples using universal culture media. ERIC molecular marker was used to study the species diversity. After clustering analysis of the species on the basis of their genetic markers, one bacterium from each cluster was selected as the representative of each cluster and then identified by DNA barcoding method using the 16srDNA. Biochemical tests were performed to confirm the molecular results. In total, 102 bacterial isolates were isolated and purified from the samples, of which only 29 isolates were extremely-halophilic. The molecular diversity of isolates, based on ERIC molecular marker, showed that isolates can be assigned to five different clusters. Five isolates selected from each cluster were selected and their 16SrDNA region were amplified and sequenced with 16SrDNA-specific primers. The results showed that the five selected isolates with 99% similarity belonged to the species Microbulbifer halophilus, Halomonas salina, Bacillus sonorensis, Salinivibrio costicola and Bacillus aquimaris. The results of molecular identification were consistent with the results of biochemical tests.

Public health is at risk because Streptococcus agalactiae is increasingly linked to incidences of bovine mastitis in Egypt. In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) were used to explore the genotyping of several antibiotic resistant Streptococcus agalactiae isolated from bovine mastitis and retail milk. Also, antimicrobial resistance genes were detected. Two hundred and sixty-six (46.7%) strains were isolated from samples of milk obtained from cases of bovine mastitis in dairy farms and retail markets representing 34 (5.96%) Streptococcus agalactiae and 232 (40.7%) other Streptococcus species strains based bacterial identification. By using of disc diffusion assays, it was examined the susceptibility of all Streptococcus isolates to twelve antimicrobial agents. The highest prevalence of resistance of Streptococcus species was observed against ampicillin (65.5%), amoxicillin (56%), tetracycline (52.5%), ofloxacin (47.8%), and nitrofurantoin (46.9%). High proportion of the S. agalactiae isolates were resistance to amoxicillin (83%), followed by tetracycline (82.4%), nitrofurantoin (64.7%), azithromycin (61.8%), and ampicillin (50%). Most (88.2%) of S. agalactiae showed multiple antibiotic resistances (MAR) phenotypes with MAR index of 0.1-0.7 and 28 different MAR patterns. The results of genetic antimicrobial resistance of S. agalactiae strains revealed amplification of blaTEM (23%) and blaCTX (26%) genes in β-lactam-resistant strains, erm(B) gene (20%), mef(A) (35%) in macrolides resistant isolates, and tet(M) in (44%) tetracycline resistant strains. Using ERIC-PCR, the present study showed the genetic diversity and heterogeneity among S. agalactiae strains (n=34) that were classified into 19 distinct ERIC-PCR groups (A–S). Between them, ERIC O (20.6%, 7) was the most widespread group. These results indicated that milk samples served as source of MAR S. agalactiae, consequently posing threats to public health, so the improvement of the hygiene regimen on the farms and promotion of the wise use of antimicrobials are necessary. The obtained findings showed that milk samples were a source of MAR S. agalactiae, endangering public health. As a result, it is essential to enhance farm hygiene practices and promote the responsible use of antimicrobials. Additional research on the epidemiology of S. agalactiae is required to add bacterial genetic information in order to help in rational vaccine strategy in the future.

Chickpea (Cicer arietinum L.) is one of the major sources of dietary protein for the majority of populations in developing countries like India. It also increases soil fertility through its symbiotic nitrogen fixation in association with Mesorhizobium ciceri. In the present study, 40 M. ciceri isolates were recovered from root nodules and the rhizospheric soil of chickpea collected from different chickpea-producing areas of the mid-Gangetic Plains, Uttar Pradesh, India. The isolates were characterized on the basis of biochemical characteristics as well as 16S rDNA sequences. All the isolates were investigated for stress tolerance and plant growth-promoting (PGP) characteristics. M. ciceri isolates have shown wide diversity in their tolerance to high temperatures, salinity and extreme pH. Large numbers of these isolates showed an ability of mineral mobilization, i.e. phosphate (55%) and zinc (42%). Also M. ciceri isolates exhibited PGP characteristics such as production of siderophores (27.5%), ammonia (20%), hydrogen cyanide (10%) and indole acetic acid (5%). Furthermore, a high genetic diversity among the isolates was observed based on the enterobacterial repetitive intergenic consensus-PCR amplified band patterns. In conclusion, these stress-tolerant PGP isolates of M. ciceri may be very crucial for the chickpea production in sustainable agriculture. [ABSTRACT FROM AUTHOR]

In May 2019, a two‐year‐old pomegranate orchard cultivar "Wonderful", located in Central Italy, showed extensive black pitting and longitudinal cankers on twigs as well as apical leaf wilting. Bacterial isolates were identified by applying biochemical, molecular and pathogenicity assays. All isolates showed identical biochemical characteristics typical of Pseudomonas syringae. In addition, upon artificial inoculations, the isolates caused symptoms in pomegranate twigs and leaves similar to those observed in the field, and in lemon fruit and pear leaves. The isolates possess the syrB gene. An ERIC‐PCR fingerprinting analysis revealed an identical pattern for the strains isolated from pomegranate and other strains previously obtained from lemon and chickpea, in Tunisia and Italy, respectively, as well as a similarity with other strains obtained from different host plants. The 16S rRNA gene analysis showed a very high similarity with other P. s. pv. syringae strains. The study allowed to conclude that the disease was caused by P. s. pv. syringae, reported for the first time as a pathogen for pomegranate. Frost and high humidity could have favoured the outbreak. The year after the outbreak, a formulate that contains zinc, copper, manganese, chitosan and garlic powder, effectively reduced the severity of the disease in the field. [ABSTRACT FROM AUTHOR]

Toxigenic and non-toxigenic V. cholerae strains can be monitored for changes in clones or serogroups, linkages between clinical and environmental isolates, genesis and clonal selection of epidemic strains, and population structure. Also, determining genetic relatedness among V. cholerae strains is critical for determining population genetic structure and evolutionary trends. In collaboration with the Sarawak Government Hospital, the present work was carried out on a total of 16 V. cholerae isolates in order to determine the genetic relatedness or heterogeneity of V. cholerae isolates from a foodborne outbreak among guests who attended a wedding ceremony in Limbang, Sarawak, Malaysia. The random amplified polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC) were conducted to compare and determine strains and trace disease-causing microorganism. The RAPD fingerprinting was conducted using a total of 10-mer oligos 100 nmole random primers (OPAE1 - OPAE20). The primers OPAE7, OPAE10, OPAE14, and OPAE17 were selected because they were the most stable and discriminatory for V. cholerae. The PCR fingerprinting of ERIC-PCR was carried out using primer set of ERIC, ERIC1R (5'-ATGTAAGCTCCTGGGGATTCAC-3'), and ERIC2F (5'-AAGTAAGTGACTGGGGTGAGCG-3'). As a result, the 1 confirmed V. cholerae O1 samples were successfully fingerprinted. Based on the profiling results, the genetic fingerprint of some of the isolates from the clinical and environmental samples had 100% similarity, as indicated by the dendrogram. This indicated that the strains shared the same genetic profile. The smaller the genetic distance, the more homogeneous the strains are. The clinical and environmental strains shared some genetic characteristics. As indicated by the dendrogram, some strains were found to be closely linked to one another, while others were heterogeneous. Therefore, RAPD-PCR and ERIC-PCR produced the highest discrimination index. By combining these typing methods, evaluation of isolates' genetic diversity may be improved. The findings of the present work demonstrated the need for continued surveillance of V. cholera in Sarawak, Malaysia. [ABSTRACT FROM AUTHOR]

During surveys in 2018 and 2019, diseased plant materials (leaves and fruits) were collected from commercial hazelnut orchards in Samsun, Ordu, and Giresun provinces in the Black Sea region of Turkey. Bacterial isolation was performed from 188 symptomatic materials and 166 bacterial colonies with yellow color were purified on GYCA plates. Twenty-eight strains were found capable of causing lesions on bean pods, hypersensitive reaction to tobacco leaves, and disease symptoms on hazelnut seedlings. They were initially identified as X. arboricola by physiological, biochemical, and MALDI-TOF MS analyses. In PCR analysis, the strains with primers XarbQ-F/XarbQ-R and XapY17-F/XapY17-R produced a single band of 402 and 943 bp, respectively. All strains showed homogeneous band profiles for ERIC-PCR. The partial DNA sequences showed that 16 strains had more than 99% gyrB sequence similarity to strains of Xanthomonas arboricola, including the pathotype strain CFBP1159PT. The rpoD sequences were found to be highly capable of distinguishing the strains from the closely related pathovars junglandis and pruni, which showed 99.2% identity with the complete genomes of X. arboricola pv corylina strains available in Genbank. Phylogenetic tree analysis using rpoD sequences allowed separation of the hazelnut strains from other pathovars of X. arboricola. Twenty-five of the 28 strains grew on nutrient medium supplemented with 2.56 mM copper sulfate, and all were able to grow on 5 ppm streptomycin sulfate. To our knowledge, this is the most comprehensive study to characterize Turkish X. arboricola pv. corylina strains using molecular identification techniques and to determine responses to copper and streptomycin. [ABSTRACT FROM AUTHOR]

Bacterial pathogens are major etiology of infectious diseases of fish. In this study we were isolated and identified the most important Gram negative bacteria as Aeromonas spp,Coliforms.,Salmonella spp.,Pseudomonas spp.,Vibrio spp.from gills,skin,intestine,kidneys of diseased fish mainly in Tilapia niloticus and environmental water at Mahmoudia canal in Elmahmoudia city,Edfina city and spans as abrakish water,and assessment of genetic and biochemically relatiotions of the prominent bacterium. Bacteria as Aeromonas spp,Coliforms.,Salmonella spp.,Pseudomonas spp., were identified phenotypically,except V.parahaemolytica spp which was the prominent bacterial species isolated was confirmed moleculary by PCR,typed it’s strains from fish and water by ERIC_PCR.inorder to answer the question wither Vibrio spp infiction considered water borne disease or not.Unfortionately ERIC_PCR nor answered as the bacterium have diverse genetic heterogenus where the isolated were similar but genetically differed . However.in most cases, the origin of the infiction is suspected to be environmental due to phenotype similarity of isolates . The aim of this study was to investigate similarity of certain bacteria in clinically diseased fish and environmental water. [ABSTRACT FROM AUTHOR]

Salmonella enteritidis (S. enteritidis) is considered a major cause of human salmonellosis and the infection is acquired through the consumption of contaminated meat and meat products with this pathogen. The objective of this study was to investigate the presence of virulence and resistance-associated genes in S. enteritidis isolates from local and imported raw chicken meat and to study their genetic diversity using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) profile analysis. The results of the current study showed that the prevalence of virulence genes was high and the isolates harbored more than five class of virulence-associated genes and their frequency were different among the isolates. All the isolates showed 100% positivity for the invite and hilA genes. Genes including lpfA, avrA, sopE2, spvB, sifA and flaK were in 91%, 82%, 82%, 82%, 73% and 54.5% respectively. The sipA was only present in one isolate. The susceptibly test showed 100% sensitivity to chloramphenicol, norfloxacin, ceftriaxone, and cefotaxime and all the isolates were 100% resistant to ampicillin. However, two of the isolates showed resistance to gentamycin and trimethoprim with sulphamethoxazole. The isolates were found to harbor the dfrA1 for trimethoprim, blaTEM for ampicillin, and the resistance rate was 100%. However, none of the S. enteritidis isolates were found to contain the resistance genes specific to fluoroquinolone, erythromycin, chloramphenicol, cefotaxime, and carbapenems (NDM and OXA) resistance. ERIC-PCR allowed the grouping of the isolates into 5 different ERIC patterns including ET1-ET5. It can be concluded that the detection of the high frequency of virulence genes and MDR suggests the high virulence potential of S. enteritidis isolates which could be of major public health concern.

Acinetobacter baumannii has recently been known as a major cause of hospital- and community-acquired infections. Carbapenem resistant A. baumanni (CRAB) has been recorded to be resistant to nearly all antibiotics, including the last resort antibiotics; carbapenems. This study aimed to detect the carbapenem resistance levels and mechanisms, in addition to the genotyping of A. baumanni in Upper Egypt. About 200 clinical samples were collected from different wards of Sohag University Hospital, Egypt, from which 20 A. baumannii isolates were recovered and then identified using conventional methods and Polymerase Chain Reaction (PCR). Antibiotic sensitivity testing was carried out using the Disk diffusion method, followed by PCR testing of the common carbapenemase-encoding genes, including OXA-51, OXA-58, KPC, GES, IMP, NDM, VIM, SIM, and GIM. Genotyping was performed using the Enterobacterial Repetitive Intergenic Consensus-Polymerase chain reaction (ERIC-PCR). About 85 % of A. baumannii strains were multidrug resistant (MDR), and high rate of Extreme drug resistant (XDR) A. baumannii (70 %) was detected. Carbapenem resistance was detected in 65 % of A. baumannii isolates, 70.58 % of MDR isolates, and 85.7 % of XDR isolates, respectively. Carbapenemase- encoding genes, including blaOXA-51, VIM, NDM, and GES, were detected in 100 %, 100 %, 76.9 2 % and 76.92 % of the carbapenem resistant A. baumannii (CRAB) isolates, respectively. The blaIMP and blaKPC genes had lower prevalence rates of 15.38 % and 30.77 %; respectively, whereas the SIM, GIM, and OXA-58 genes were not detected in any of the tested A. baumanni isolates. All of the MDR isolates carried three or more the carbapenemases encoding genes, and 85.7 % of the XDR isolates carried four or more of the carbapenemase-encoding genes. The dendrogram constructed from the ERIC-PCR results showed that the A. baumannii isolates were divided into three different clusters.

Lamya Zohair Yamani, Nasreldin Elhadi Department of Clinical Laboratory Science, College of Applied Medical Sciences, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom of Saudi ArabiaCorrespondence: Nasreldin Elhadi, Department of Clinical Laboratory Science, Imam Abdulrahman Bin Faisal University, Dammam, Kingdom of Saudi Arabia, Email nmohammed@iau.edu.saIntroduction: The Kingdom of Saudi Arabia has limited data on enteropathogenic Escherichia coli (EPEC). Therefore, this study was undertaken to contribute to EPEC surveillance and investigate the molecular epidemiology of EPEC strains that have been implicated in human infection in King Fahd Hospital of the University (KFHU) between 2013 and 2014 in the Eastern Province of Saudi Arabia.Methods: A total of 60 non-duplicate E. coli isolates associated with human gastroenteritis were included in this study. They were characterized using PCR to determine virulence genes, antimicrobial resistance patterns, and enterobacterial repetitive intergenic consensus (ERIC-PCR).Results: Among the 60 strains, 20% of those examined were positive for the intimin eae and bfpA genes and identified as typical EPEC (tEPEC). Furthermore, 44 of E. coli strains tested positive for the eae gene only and revealed a high occurrence rate of 73.3% of atypical EPEC (aEPEC) within the overall examined strains. All strains were positive for the EAST1 gene, and none tested positive for the stx gene. More than 70% of EPEC strains were multi-drug resistant (MDR) and aEPEC strains with the highest proportion of this feature of MDR. ERIC-PCR fingerprint revealed a total of 19 ERIC types with eight related distinct clusters and a similarity rate cut-off with ≥ 90% homology from the identified isolates.Conclusion: A high antibiotic resistance rate was reported for first-line antibiotics, such as ampicillin, tetracycline, nalidixic acid, trimethoprim-sulfamethoxazole, noroxin, and ciprofloxacin.Keywords: EPEC, virulence, antimicrobial resistance, MDR, ERIC-PCR

Abstract There is a global concern and increasing reports regarding foodborne disease infections associated with consuming contaminated vegetables, seafood, meat, and poultry products. Among foodborne bacterial pathogens globally, Salmonella, Escherichia coli, and Shigella were the most frequently implicated in causing food poisoning infections in children and adults. In Saudi Arabia, the consumption rates of imported fresh fruits, vegetables, seafood, and meat products are considered high. Therefore, the development of simple PCR based DNA fingerprinting methods is essential to track the source and route of microbial contamination among imported frozen meat and seafood products. A total of 38 E. coli strains were subtyped using ERIC1R, ERIC2, and a pair combination (ERIC1R + ERIC2) to generate genomic fingerprinting. The three Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR)-based primers were generated in 26, 24, and 16 different genotypes while using ERIC1R, ERIC2, and ERIC1R + ERIC2, respectively. The Discrimination Index values obtained by ERIC1R, ERIC2, and ERIC1R + ERIC2 were 0.976, 0.965, and 0.903, respectively. ERIC1R and ERIC2 primers had the best discriminatory ability and typeability value and proved suitable for investigating genetic analysis among the population of E. coli strains. At the same time, the ERIC1R + ERIC2 primer pair has average discriminatory power and typeability value for differentiating E. coli strains. These results suggest that subtyping using ERIC1R and ERIC2 primer is a more reliable and rapid typing strategy for E. coli strains.