The Value of Preoperative Ultrasound-Determined Fluid Film and Joint Aspiration in Revision Hip Arthroplasty.

BACKGROUND: Data regarding the diagnostic value of ultrasound (US)-determined fluid film and joint aspiration prior to revision total hip arthroplasty for suspected periprosthetic joint infections (PJIs) are limited. This study aimed to analyze the value of US-determined fluid film, characterized the preoperative and intraoperative microbiological spectrum and resistance patterns, and compared the concordance between preoperative synovial fluid and intraoperative culture results. METHODS: We analyzed 366 US examinations from 324 patients prior to revision total hip arthroplasty. Selected cases were grouped into clearly infected, noninfected, and inconclusive cohorts, according to the International Consensus Meeting 2018 Criteria. For US-determined fluid film <1 mm, no aspiration was performed based on our institutional protocol. Patients were grouped into no aspiration (144 of 366; [39.3%]), dry tap (21 of 366; [5.7%]), and a successful tap (201 of 366; [54.9%]). The microbiological spectrum and antibiograms were compared between preoperative and intraoperative results. RESULTS: The absence of US-determined fluid film showed no correlation with the presence of a hip PJI. Overall, 31.9% cases of the no-aspiration group had a PJI. In total, 13.5% discrepancies were found between successful taps and intraoperative cultures. The most prevalent microorganisms in preoperative synovial fluid were Staphylococcus epidermidis and Staphylococcus aureus (20.8%), while intraoperatively S. epidermidis (26.3%) and Cutibacterium acnes (14.5%) were leading. Additional microorganisms were identified in 32.5% of intraoperative cultures. There were no differences between resistance patterns of preoperative and intraoperative concordant microorganisms. CONCLUSIONS: Absence of US-determined fluid film cannot rule out the presence of a hip PJI. Combined microbiological results from hip US aspirations and subsequent surgical procedures are crucial to design an effective treatment for suspected hip PJI.

Sociodemographic distributions and molecular characterization of colonized Enterococcus faecium isolates from locality hospitals in Khartoum, Sudan.

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.

Surveillance Study of Klebsiella pneumoniae in the Giant Panda Revealed High Genetic Diversity and Antibiotic Therapy Challenge.

Klebsiella pneumoniae is not only a worldwide human pathogen, it also effects wildlife, such as the giant panda (Ailuropoda melanoleuca), in which it has recently been evidenced to result in diarrhea, organ failure, and even death. A K. pneumoniae investigation was carried out at the Chengdu Research Base of Giant Panda Breeding in 2018. As part of the investigation, the pulsed-field gel electrophoresis (PFGE) typing, multilocus-sequence typing (MLST), antibiotic resistance profiles (ARPs), and antibiotic resistance genes (ARGs) were studied based on all isolates. Fecal samples were collected from 72 A. melanoleuca from May to December 2018, and a total of 90 K. pneumoniae were isolated from 153 fecal samples. The genotyping results showed that the isolates had high diversity, of which 84 clusters were obtained by PFGE and 57 STs by MLST. The overall trend of the similarity of isolates was the first sample period > second sample period > third sample period, which showed the increasement of genome variability of K. pneumoniae. In addition, 90 isolates showed high resistance to ampicillin, rifampicin, and compound sulfamethoxazole. Of the obtained isolates, 50% carried 6~8 ARPs, and the carrying volume increased during three sample periods, in which we found two isolates carrying 12 and 13 ARPs during the third sample period, respectively. Moreover, a total of 65 ARGs were detected (90.28%, 65/72) in 90 K. pneumoniae samples. Almost all bacteria sampled contained 17 ARGs that belonged to the ß-lactamase, Multidrug, MGEs, Aminoglycoside, and Tetracycline, which may be the basis of ARPs of K. pneumoniae. Moreover, the types of Multidrug and MGEs had a greater impact on antibiotic susceptivity of K. pneumoniae. Our results showed that K. pneumoniae has a serious risk of transmission in A. melanoleuca and K. pneumoniae had a high possibility of genome diversity and the risk of drugs tolerance under the large antibiotic usage.

Prevalence and Antibiotics Susceptibility Pattern of Urine Bacterial Isolates from Tripoli Medical Center (TMC), Tripoli, Libya / Prevalencia y patrón de susceptibilidad a los antibióticos de los aislados bacterianos en orina del centro médico de Trípoli (TMC), Trípoli, Libia

Introduction: Urinary tract infections (UTI) are one of the most common human bacterial infections encountered by physicians. The risk of resistant microbes is emerging as a result of the overuse of antibiotics treatments. The presence of pathogens with increased resistance to antimicrobial agents makes UTIs difficult to treat. This study was aimed at determining the prevalence of the pathogens that cause UTIs, as well as the antibiotic susceptibility of these isolates.Materials and methods: This prospective study was conducted from February 2020 to April 2020; a total number of 200 urine samples were collected from patients who daily attended TMC Libya. Bacterial pathogens were determined by bacteriological culture methods and Antimicrobial susceptibility testing was done by using the disc diffusion method.Results: Out of 200 samples, 110 cases had a positive culture. The dominant bacterial pathogens were Gram-negative that being with Escherichia coli (49, 55.68%), followed by Klebsiella pneumonia (18, 20.46%), Pseudomona aeruginosa (9, 10.23%), Proteus mirabilis (8, 9.09%), Enterobacter aerogenes (2, 2.27%), Citrobacter freundii (2, 2.27%). Gram-positive bacteria were Staphylococcus aureus 20 (90.91%) followed by S. saprophyticus (2, 9.01%) of the isolate’s strains. The isolated uropathogen showed increased levels of resistance to antibiotics. Where the Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus indicated the highest antibiotic resistance to Nitrofurantoin, Sulfamethoxazole/trimethoprim, Tetracycline, Ciprofloxacin, Metronidazole and also revealed the most sensitivity to Cefixime followed by doxycycline and ceftriaxone.Conclusions: The obtained results emphasized the emergence of highly resistant bacteria to most of the tested antimicrobials and propose the need for physicians to change their treatment pattern depending on antimicrobial susceptibility results.(AU)

Introducción: Las infecciones del tracto urinario (ITU) son una de las infecciones bacterianas humanas más comunes que encuentran los médicos. El riesgo de microbios resistentes está surgiendo como resultado del uso excesivo de tratamientos con antibióticos. La presencia de patógenos con mayor resistencia a los agentes antimicrobianos dificulta el tratamiento de las infecciones urinarias. Este estudio tuvo como objetivo determinar la prevalencia de los patógenos que causan las ITU, así como la susceptibilidad a los antibióticos de estos aislados.Materiales y métodos: Este estudio prospectivo se realizó desde febrero de 2020 hasta abril de 2020; Se recogió un total de 200 muestras de orina de pacientes que asistían diariamente a TMC Libia. Los patógenos bacterianos se determinaron mediante métodos de cultivo bacteriológico y las pruebas de susceptibilidad a los antimicrobianos se realizaron mediante el método de difusión en disco.Resultados: De 200 muestras, 110 casos tuvieron un cultivo positivo. Los patógenos bacterianos dominantes fueron Gram-negativos con Escherichia coli (49, 55.68%), seguidos de Klebsiella pneumonia (18, 20.46%), Pseudomonas aeruginosa (9, 10.23%), Proteus mirabilis (8, 9.09%), Enterobacter aerogenes (2, 2.27%). %), Citrobacter freundii (2, 2.27%). Las bacterias grampositivas fueron Staphylococcus aureus (20, 90.91%) seguido de S. saprophyticus, (2, 9.01%) de las cepas del aislado. El uropatógeno aislado mostró niveles aumentados de resistencia a los antibióticos. Donde Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus indicaron la mayor resistencia a los antibióticos a la nitrofurantoína, sulfametoxazol/trimetoprima, tetraciclina, ciprofloxacina, metronidazol y también revelaron la mayor sensibilidad a la cefixima seguida de la doxiciclina y la ceftriaxona.Conclusiones: Los resultados obtenidos enfatizan la aparición de bacterias altamente resistentes a la mayoría de los antimicrobianos probados...(AU)

Antimicrobial Resistance and Genotyping of Bacteria Isolated from Urinary Tract Infection in Children in an Iranian Referral Hospital.

INTRODUCTION: Urinary tract infection (UTI) is one of the most common bacterial infections in childhood, and the increasing rate of antibiotic resistance to the commonly prescribed antimicrobial agents against it has become a major concern. The aim of this study was to determine the antibiotic resistance and genotyping of bacteria isolated from urine cultures in patients referred to the Children's Medical Center, Tehran, Iran. METHODS: During the 1-year period, antimicrobial susceptibility profiles of strains isolated from patients with UTI were determined. Typing of the isolates causing nosocomial infections was performed by random amplified polymorphic DNA (RAPD) analysis, and the results were analyzed by Gelcompar II software. RESULTS: In this study, 203 children (130 girls and 73 boys) were included. The patients' age ranged from 1 day to 16 years (IQR average=4 months to 4 years). The most frequent isolated organisms were Escherichia coli (118 isolates, 58%), followed by Klebsiella pneumoniae (30 isolates, 15%). Sixty-two strains (18 strains of E. coli, 13 strains of K. pneumoniae, 11 strains of Enterococcus faecium, and five strains of Burkholderia cepacia complex) had criteria of nosocomial infection. A high resistance rate to trimethoprim-sulfamethoxazole (69%) and cefotaxime (60%) was reported in E. coli and K. pneumoniae strains, respectively. Pseudomonas aeruginosa strains showed high sensitivity to amikacin (100%). All E. faecium strains were susceptible to trimethoprim-sulfamethoxazole (100%), and 23% of the strains were resistant to vancomycin. The analysis of RAPD-typing revealed the presence of three clusters in E. coli, two clusters in E. faecium, and one clone in K. pneumoniae. Besides, four out of five isolates of B. cepacia complex had more than 90% genetic similarity. CONCLUSION: The most frequent isolated pathogen was E. coli, and an increasing rate of antibiotic resistance to the commonly prescribed antimicrobial agents such as trimethoprim/sulfamethoxazole and cephalosporins was observed. Moreover, the results of this study showed the presence of clones with ≥80% similarity in E. coli, K. pneumoniae, E. faecium, and B. cepacia complex isolates; therefore, the transmission of nosocomial infections from one patient to another or one ward to another is probable.

Three Year Study of Infection Profile and Antimicrobial Resistance Pattern from Burn Patients in Southwest Iran.

INTRODUCTION: Burn wound infections, as one of the most important risk factors, cause serious complications in burns. Hence, the focus of medical care should be preventing infections and resistant isolates. The current study investigates the prevalence of infectious agents and antimicrobial resistance patterns during three years. MATERIALS AND METHODS: A total of 960 isolates were collected from different sample kinds cultured for 615 burn patients who were hospitalized during January 2016 to December 2018 in Amir-Al-Momenin Burn Center. The type of microorganism and their antibiotic resistance patterns were identified by microbiological tests and the standard disk-diffusion method according to the introduced standard techniques. RESULTS: Incidence of positive growth was seen more in males than in females. Most of the burns encountered were due to flame injuries (35.4%). Based on the diversity of bacterial isolates, Pseudomonas aeruginosa was the most frequent pathogen (49.9%), followed by Klebsiella sp. (9.7%), Acinetobacter sp. (7.2%) and Staphylococcus aureus (6.5%). The trend of resistance of meropenem was declining in P. aeruginosa isolates. Klebsiella sp. as the second most prevalent agent showed a high level of resistance to the studied antibiotics. The antibiogram results for S. aureus isolates showed an increasing trend in MRSA isolates. CONCLUSION: By evaluating the infectious agent, it was found that although frequencies of microorganisms and resistant isolates were a little high, performing a multidisciplinary approach controls the trend during the study period. These achievements have been gained due to a strict politicized infection control and stewardship program in the appointed burn center.

Phenotypic and Genotypic Assessment of Antibiotic Resistance of Staphylococcus aureus Bacteria Isolated from Retail Meat.

BACKGROUND: Resistant Staphylococcus aureus (S. aureus) bacteria are determined to be one of the main causes of foodborne diseases. PURPOSE: This survey was done to assess the genotypic and phenotypic profiles of antibiotic resistance of S. aureus bacteria isolated from retail meat. METHODS: Four-hundred and eighty-five retail meat samples were collected and examined. S. aureus bacteria were identified using culture and biochemical tests. The phenotypic profile of antibiotic resistance was examined using the disk diffusion method. The genotypic pattern of antibiotic resistance was determined using the polymerase chain reaction. RESULTS: Forty-eight out of 485 (9.89%) raw retail meat samples were contaminated with S. aureus. Raw retail buffalo meat (16%) had the highest incidence of S. aureus, while raw camel meat (4%) had the lowest. S. aureus bacteria exhibited the uppermost incidence of resistance toward tetracycline (79.16%), penicillin (72.91%), gentamicin (60.41%), and doxycycline (41.666%). The incidence of resistance toward chloramphenicol (8.33%), levofloxacin (22.91%), rifampin (22.91%), and azithromycin (25%) was lower than other examined antibiotics. The most routinely detected antibiotic resistance genes were blaZ (58.33%), tetK (52.08%), aacA-D (33.33%), and ermA (27.08%). Cat1 (4.16%), rpoB (10.41%), msrA (12.50%), grlA (12.50%), linA (14.58%), and dfrA1 (16.66%) had the lower incidence rate. CONCLUSION: Raw meat of animals may be sources of resistant S. aureus which pose a hygienic threat about the consumption of raw meat. Nevertheless, further investigations are essential to understand supplementary epidemiological features of S. aureus in retail meat.

Antibiotic resistance and molecular characterization of enteroaggregative Escherichia coli isolated from patients with diarrhea in the Eastern Province of Saudi Arabia.

AIM: To investigate presence of Enteroaggregative Escherichia coli (EAEC) in patients suffering with diarrhea by targeting the pCVD432 (pAA) gene using PCR. METHODS: There were 63 non-duplicate isolates of E. coli isolated from diarrheal cases in teaching hospital in Eastern Province of Saudi Arabia between May 2013 to July 2014. All E. coli strains were examined for antibiotic susceptibility testing and polymerase chain reaction (PCR) for detection of virulence gene markers for EAEC. RESULTS: Of the 63 E coli strains that were reported with diarrheal cases, 35 (55.6%) EAEC were tested positive for pCVD432 gene and aggR gene was present in 19 (54.3%) strains. All strains tested positive for pCVD432 and aggR genes were classified as typical EAEC (tEAEC). EAEC revealed resistance to tetracycline, ampicillin, nalidixic acid, trimethoprim sulfamethoxazole, ciprofloxacin, streptomycin, noroxin, and piperacillin. CONCLUSION: EAEC was detected for the first time, among Saudi patients with diarrhea in this region of Saudi Arabia. The reported antibiotic resistance in this study is considered high among isolated EAEC strains to routinely prescribed antibiotics in our area.

Meta-Analysis of Biofilm Formation, Antibiotic Resistance Pattern, and Biofilm-Related Genes in Pseudomonas aeruginosa Isolated from Clinical Samples.

Resistant microorganisms such as Pseudomonas aeruginosa grow by developing biofilms in hospitals. We aimed to investigate the biofilm formation and the frequencies of biofilm-related genes and their associations with antibiotic resistance pattern in P. aeruginosa isolated from Iranians' clinical samples. This review was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We conducted a systematic literature search in scientific databases using medical subject heading terms, including "Pseudomonas aeruginosa," "biofilm formation," "biofilm-related genes," "antibiotic resistance," and "prevalence," to obtain related articles published from 1st January, 2000, to 30th March, 2019. The studies reporting the prevalence of biofilm formation, the frequencies of biofilm-related genes, and the antibiotic resistance pattern in P. aeruginosa retrieved from Iranian patients were included. Meta-analysis was performed using the Comprehensive Meta-Analysis software. The pooled rate of biofilm formation was calculated as 86.5% (95% confidence interval [CI]: 79-91.6). The combined frequencies of strong, moderate, and weak biofilms were 51% (95% CI: 37.4-64.4), 29.2% (95% CI: 20.9-39.1), and 25.4% (95% CI: 11.5-47.2), respectively. The pooled prevalence of laslR, algD, algU, ppyR, and pelF genes were 93.6% (95% CI: 88.1-96.6), 91.4% (95% CI: 80.8-96.4), 89.3% (95% CI: 85.2-92.3), 98.7% (95% CI: 96.5-99.6), and 93% (95% CI: 82.7-97.3), respectively. The highest combined antibiotic resistance rates of P. aeruginosa isolates were against piperacillin/tazobactam (90%). This study showed that biofilm formation was higher in multidrug-resistant (MDR) P. aeruginosa than non-MDRs. A significant correlation was observed between biofilm formation and antibiotic resistance in 50% of studies included in this review.

Phenotypic and molecular characterization of antimicrobial resistance in Trueperella pyogenes strains isolated from bovine mastitis and metritis.

BACKGROUND: Trueperella pyogenes is one of the most clinically imperative bacteria responsible for severe cases of mastitis and metritis, particularly in postpartum dairy cows. The bacterium has emergence of antibiotic resistance and virulence characters. The existing research was done to apprise the phenotypic and genotypic evaluation of antibiotic resistance and characterization of virulence factors in the T. pyogenes bacteria of bovine mastitis and metritis in postpartum cows. METHODS: Two-hundred and twenty-six bovine mastitic milk and 172 uterine swabs were collected and transferred to laboratory. Samples were cultured and T. pyogenes isolates were subjected to disk diffusion and DNA extraction. Distribution of virulence and antibiotic resistance genes was studied by PCR. RESULTS: Thirty-two out of 226 (14.15%) mastitic milk and forty-one out of 172 (23.83%) uterine swab samples were positive for T. pyogenes. Isolates of mastitic milk harbored the highest prevalence of resistance toward gentamicin (100%), penicillin (100%), ampicillin (90.62%), amoxicillin (87.50%) and trimethoprim-sulfamethoxazole (87.50%), while those of metritis harbored the highest prevalence of resistance toward ampicillin (100%), amoxicillin (100%), gentamicin (97.56%), penicillin (97.56%) and cefalexin (97.56%). AacC, aadA1, aadA2 and tetW were the most generally perceived antibiotic resistance genes. All bacteria harbored plo (100%) and fimA (100%) virulence factors. NanP, nanH, fimC and fimE were also the most generally perceived virulence factors. CONCLUSIONS: All bacteria harbored plo and fimA virulence factors which showed that they can use as specific genetic markers with their important roles in pathogenicity of T. pyogenes bacteria. Phenotypic pattern of antibiotic resistance was confirmed by genotypic characterization of antibiotic resistance genes.

Acinetobacter baumannii in sheep, goat, and camel raw meat: virulence and antibiotic resistance pattern.

Acinetobacter genus belongs to a group of Gram-negative coccobacillus. These bacteria are isolated from human and animal origins. Antimicrobial agents play a vital role in treating infectious diseases in both humans and animals, and Acinetobacter in this regard is defined as an organism of low virulence. The current study aimed to evaluate antibiotic resistance properties and virulence factor genes in Acinetobacter baumannii strains isolated from raw animal meat samples. Fresh meat samples from 124 sheep, 162 goat, and 95 camels were randomly collected from Isfahan and Shahrekord cities in Iran. Most A. baumannii strains isolated from sheep meat samples represented fimH (82.35%), aac(3)-IV (78.43%), sul1 (78.43%) and Integron Class I (96.07%) genes. Moreover, more than 50% of A. baumannii strains isolated from sheep samples were resistant to streptomycin (54.90%), gentamycin (74.50%), co-trimoxazole (70.58%), tetracycline (82.35%), and trimethoprim (62.74%). Current findings revealed significant association between the presence of fimH, cnfI, afa/draBC, dfrA1, sulI, aac(3)-IV genes in sheep samples. Furthermore, significant association was observed between fimH, cnfI, sfa/focDE and dfrA1genes in goat meat samples. In sheep meat samples, significant differences were identified in resistance to gentamicin, tetracycline, and co-trimoxazole in comparison with other antibiotics. Finally, there were statistically significant differences between the incidences of resistance to gentamicin, tetracycline, and co-trimoxazole in comparison with other antibiotics in all strains. In conclusion, the presence of virulence factors and antibiotic resistance in A. baumannii strains isolated from animal meat samples showed that animals should be considered as a potential reservoir of multidrug-resistant A. baumannii.

Molecular characterization, serotypes and phenotypic and genotypic evaluation of antibiotic resistance of the Klebsiella pneumoniae strains isolated from different types of hospital-acquired infections.

Purpose: Virulent and resistant Klebsiella pneumoniae strains are considered as one of the most significant causes of hospital-acquired infections. The present investigation was done to study the distribution of virulence factors, capsule serotypes and phenotypic and genotypic evaluation of antibiotic resistance of the K. pneumoniae strains isolated from hospital-acquired infections. Patients Materials and methods: Two hundred and sixty different types of hospital-acquired infections were collected and cultured. Antibiotic resistance pattern of K. pneumoniae isolates and their molecular characterization were studied using disk diffusion and PCR, respectively. Results: One hundred and fifty out of 260 (44.22%) hospital-acquired infections harbored K. pneumoniae. Urine samples (63.75%) had the highest prevalence of K. pneumoniae, while wound (33.33%) had the lowest. K. pneumoniae strains harbored the highest prevalence of resistance against ampicillin (100%), cefuroxime (100%), amoxicillin/clavulanic acid (95.65%) and ceftazidime (95.52%). FimH-1 (93.04%), traT (92.17%), mrkD (84.34%), and entB (80.86%) were the most commonly detected virulence genes. AcrAB (96.52%) and tolC (85.21%) were the most commonly detected antibiotic resistance genes. Prevalence of ompK35 and ompK36 virulence genes were 75.65% and 79.13%, respectively. Prevalence of K1 and K2-positive serotypes were 27.82% and 6.96%, respectively. Conclusions: High prevalence of resistance against several types of antibiotics and simultaneous presence of some virulence factors and multi-drug resistance genes pose an important public health issue.

Genotyping and distribution of putative virulence factors and antibiotic resistance genes of Acinetobacter baumannii strains isolated from raw meat.

Background: Acinetobacter baumannii strains with multiple antimicrobial resistance are primarily known as opportunistic nosocomial bacteria but they may also be regarded as emerging bacterial contaminants of food samples of animal origin. Here we aimed to study the molecular characteristics of the A. baumanni strains isolated from raw meat samples. Methods: A total of 22 A. baumanni strains were isolated from 126 animal meat samples and were genotyped by ERIC-PCR method and by PCR detection of their virulence and antimicrobial resistance determinants. A. baumannii strains with 80% and more similarities were considered as one cluster. Results: Sixteen different genetic clusters were found amongst the 22 A. baumanni strains. Of the 22 strains, 12 (54.54%) had similar genetic cluster. A. baumannii strains exhibited the highest percentage of resistance against tetracycline (90.90%), trimethoprim (59.09%), cotrimoxazole (54.54%) and gentamicin (50.00%). TetA (81.81%), tetB (72.72%), dfrA1 (63.63%), aac(3)-IV (63.63%), sul1 (63.63%) and aadA1 (45.45%) were the most commonly detected antibiotic resistance genes. FimH (81.81%), afa/draBC (63.63%), csgA (63.63%), cnf1 (59.09%), cnf2 (54.54%) and iutA (50.00%) were the most commonly detected virulence factors. A. baumannii strains isolated from the chicken meat samples had the highest similarities in the genetic cluster. Conclusions: A. baumannii strains with similar genetic cluster (ERIC-Type) had the same prevalence of antibiotic resistance, antibiotic resistance genes and virulence factors. Genetic cluster of the A. baumannii strains is the main factor affected the similarities in the genotypic and phenotypic properties of the A. baumannii strains.

Biofilm testing of microbiota: An essential step during corneal scrap examination in Egyptian acanthamoebic keratitis cases.

PURPOSE: To detect co-infections in the culture-proven acanthamoebic keratitis (AK) cases, and to test the capability of biofilm formation in the isolated microbiota. The clinical findings, habit of wearing contact lens and in-vitro antibiotic resistance were analyzed further according to the biofilm formation capability. METHODS: After clinical examination, corneal scraps and swabs were taken from 240 clinically suspected AK cases, for Acanthamoeba and microbiological cultures. In cases of keratoplasty, trimmed corneal tissue was collected and sent for histopathological examination. Scanning electron microscopy was done for some samples. Biofilm formation capability was investigated using a tissue culture plate method. Antibiotic resistance pattern was determined using a modified-Kirby-Bauer disc diffusion method. RESULTS: In 102 AK culture proven cases, 11 had no co-infection, 74 had a single co-infection and 17 had double co-infections. Enterobactericae and Aspergillus were the commonest bacterial and fungal isolates, respectively. Regarding the biofilm formation, 64.7% of Enterobactericae, 50% of Pseudomonas aeuroginosa, 43.75% of Staph aureus, 76.92% of Streptococcus pneumoniae, 28.57% of Corynebacterium, 60% of α-haemolytic streptococci, 40% of Acinetobacter, 100% of Candida and 77.8% Aspergillus isolates were biofilm producers. Severe manifestations were more frequently reported in cases co-infected with biofilm producers than with non-biofilm producers. Generally, high percentages of the biofilm forming bacterial isolates were sensitive to antibiotics in-vitro. CONCLUSIONS: Routine investigations for co-infection and biofilm formation in addition to Acanthamoeba culture are strongly recommended in suspected AK cases. Co-infection with biofilm producers may precipitate extrinsic in-vivo drug resistance despite of the in-vitro sensitivity. Designing a biofilm-dissolving topical drug is highly recommended to enhance the response to the standard therapeutic regimen especially in the resistant AK cases.

VacA and cagA genotypes status and antimicrobial resistance properties of Helicobacter pylori strains isolated from meat products in Isfahan province, Iran.

Although Helicobacter pylori has a significant impact on the occurrence of severe clinical syndromes, its exact ways of transmission and origin have not been identified. According to the results of some previously published articles, foods with animal origins play a substantial role in the transmission of H. pylori to humans. The present investigation was carried out to study the vacuolating cytotoxin A (vacA) and cytotoxin associated gene A (cagA) genotypes status and antibiotic resistance properties of H. pylori strains recovered from minced-meat and hamburger samples. A total of 150 meat product samples were collected from supermarkets. All samples were cultured and the susceptive colonies were then subjected to nested-PCR, PCR-based genotyping and disk diffusion methods. 11 out of 150 samples (7.33%) were positive for H. pylori. All the isolates were further identified using the nested-PCR assay. Prevalence of H. pylori in hamburger and minced-meat samples was 1.42% and 12.5%, respectively. S1a, m1a and cagA were the most commonly detected genotypes. The most commonly detected combined genotypes in the H. pylori strains of minced-meat were s1am1a (10%), s1am1b (10%) and s2m1a (10%). Helicobacter pylori strains of meat products harbored the highest levels of resistance against ampicillin (90.90%), erythromycin (72.72%), amoxicillin (72.72%), trimethoprim (63.63%), tetracycline (63.63%), and clarithromycin (63.63%). Hamburger and minced-meat samples may be the sources of virulent and resistant strains of H. pylori. Meat products are possible sources of resistant and virulent strains of H. pylori similar to those vacA and cagA genotypes. Using healthy raw materials and observation of personal hygiene can reduce the risk of H. pylori in meat products.

Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran.

BACKGROUND: Uropathogenic Escherichia coli O- Serogroups with their virulence factors are the most prevalent causes of UTIs. The present research performed to track common uropathogenic E.coli serogroups, antibiotic resistance pattern of strains and prevalence of virulence genes in isolations having the ability to constitute biofilm. METHODS: In this research 130 E.coli isolation from patients having UTI symptoms were collected and antimicrobial resistance pattern was performed by Kirby-Bauer method. Polymerase chain reaction was done using primer pairs to identify common serogroups of uropathogenic E.coli and studying virulence genes in isolations creating biofilm. RESULTS: Among 130 E.coli isolates, 80 (61.53 %) were able to make biofilm that 15 isolates (18.75 %) indicated strong reaction, 20 (25 %) of medium and 45 (56.25 %) of weak biofilm reaction. Among isolations creating biofilm, the highest resistance reported to Ampicillin (87.5 %) and the lowest to Nitrofurantoin (3.75 %). The frequency of fimH, pap, sfa and afa genes in isolations having the ability to create strong biofilm reported 93.33 %, 86.66 %, 86.66 % and 66.66 %, respectively. CONCLUSIONS: The findings indicated the importance of virulence genes in serogroups producing uropathogenic E.coli biofilm. It is recommended that strains producing biofilm before antibiotic use should be studied.

Molecular identification of Acinetobacter baumannii isolated from intensive care units and their antimicrobial resistance patterns.

BACKGROUND: Acinetobacter baumannii is one of the most important pathogens in hospital-acquired infections especially in intensive care units (ICUs). This opportunistic pathogen can be easily isolated from water, soil, and hospital facilities. A. baumannii as a nosocomial opportunistic pathogen is resistant to a wide range of antibiotics and responsible for multiple infections, including bacteremia, pneumonia, meningitis, urinary tract infections, and surgical wounds. The aim of this study was to determine frequency and resistance patterns of A. baumannii isolated in ICUs of Isfahan Hospitals. MATERIALS AND METHODS: During 1 year period (2012-2013), 350 specimens were collected from ICUs of Isfahan hospitals. The isolates were characterized as A. baumannii by conventional phenotypic, biochemical tests and confirmed by PCR for OXA-51-like gene. Susceptibility of isolates was determined by standard disk diffusion method according to CLSI. RESULTS: From total of 350 specimens, 43 isolates were A. baumannii. The antimicrobial patterns of isolates showed that 53.5% of isolates were resistant to amikacin, 83.7% to tetracyclin, 86% to ceftazidime, 90.7% to Trimethoprim sulfametoxazol, 93% to imipenem, cefepime, meropenem, ampicillin-sulbactam. All isolates were resistant to ciprofloxacin. CONCLUSION: This study showed a high resistance of A. baumannii to a wide range of antimicrobial agent. It is necessary to adopt appropriate strategies to control the spread of the bacteria in care unit centers and wards.

Antibiotic resistance pattern of Pseudomonas aeruginosa isolated from urine samples of Urinary Tract Infections patients in Karachi, Pakistan.

OBJECTIVE: The aim of this study was to evaluate the antibiotic resistance pattern of Psedomonas aeruginosa and its prevalence in patients with urinary tract infections (UTI) for effective treatment in a developing country like Pakistan. METHODS: This is an observational study conducted for a period of ten months which ended on December 2013 at the Dr. Essa Laboratory and Diagnostic Centre in Karachi. A total of 4668 urine samples of UTI patients were collected and standard microbiological techniques were performed to identify the organisms in urine cultures. Antibiotic susceptibility testing was performed by Kirby-Bauer technique for twenty five commonly used antimicrobials and then analyzed on SPSS version 17. RESULTS: P. aeruginosa was isolated in 254 cultures (5.4%). The most resistant drugs included Ceclor(100%) and Cefizox (100%) followed by Amoxil/Ampicillin (99.6%), Ceflixime (99.6%), Doxycycline (99.6%), Cefuroxime (99.2%), Cephradine (99.2%), Cotrimoxazole (99.2%), Nalidixic acid (98.8%), Pipemidic acid (98.6%) and Augmentin (97.6%). CONCLUSION: Emerging resistant strains of Pseudomonas aeruginosa are potentially linked to injudicious use of drugs leading to ineffective empirical therapy and in turn, appearance of even more resistant strains of the bacterium. Therefore, we recommend culture and sensitivity testing to determine the presence of P.aeruginosa prior to specific antimicrobial therapy.

Simultaneous Detection of Integrase and Antibiotic Resistance Genes within SXT Constin in Vibrio cholerae O1 El Tor Strains Isolated from Iran Using Multiplex-PCR.

OBJECTIVES: Amongst the various antibiotic resistant elements in Vibrio. cholerae, SXT constin (SXT-C) is important. We were going to design a quick method for determination of antibiotic resistance gene pattern in SXT-C. MATERIALS AND METHODS: Ninety four V. cholerae O1 El Tor isolates were used in this study. Antibiotic susceptibility testing, multiplex PCR amplification of SXT-C containing dfrA1, sulII, strB and the int in a multiplex PCR were done. RESULTS: Results of our study showed that 92 (97.8%) out of 94 isolates were positive for all above genes. Multiplex PCR results correlated with the antibiotic susceptibility data obtained by using disc diffusion assay. CONCLUSION: Using this multiplex PCR, it would be easily possible to demonstrate the presence of antibiotic resistance in SXT-C which, in turn, allows for a suitable treatment in cholera patients causing reduction in the mortality and morbidity rate of the infected individuals.

Identification and antimicrobial resistance of microflora colonizing feral pig (Sus scrofa) of Brazilian Pantanal

Antimicrobial resistance of bacteria is a worldwide problem affecting wild life by living with resistant bacteria in the environment. This study presents a discussion of outside factors environment on microflora of feral pigs (Sus scrofa) from Brazilian Pantanal. Animals had samples collected from six different body sites coming from two separated geographic areas, Nhecolandia and Rio Negro regions. With routine biochemical tests and commercial kits 516 bacteria were identified, with 240 Gram-positive, predominantly staphylococci (36) and enterococci (186) strains. Among Gram-negative (GN) bacteria the predominant specimens of Enterobacteriaceae (247) mainly represented by Serratia spp. (105), Escherichia coli (50), and Enterobacter spp. (40) and specimens not identified (7). Antimicrobial susceptibility was tested against 17 drugs by agar diffusion method. Staphylococci were negative to production of enterotoxins and TSST-1, with all strains sensitive towards four drugs and highest resistance toward ampicillin (17 percent). Enterococci presented the highest sensitivity against vancomycin (98 percent), ampicillin (94 percent) and tetracycline (90 percent), and highest resistance pattern toward oxacillin (99 percent), clindamycin (83 percent), and cotrimoxazole (54 percent). In GN the highest resistance was observed with Serratia marcescens against CFL (98 percent), AMC (66 percent) and AMP (60 percent) and all drugs was most effective against E. coli SUT, TET (100 percent), AMP, TOB (98 percent), GEN, CLO (95 percent), CFO, CIP (93 percent). The results show a new profile of oxacillin-resistant enterococci from Brazilian feral pigs and suggest a limited residue and spreading of antimicrobials in the environment, possibly because of low anthropogenic impact reflected by the drug susceptibility profile of bacteria isolated.