API 20E Research Articles

Caracterización fenotípica y simbiótica de las bacterias que nodulan Genista saharae en la región árida de Argelia

Twenty bacterial strains had been isolated from root nodules of Genista saharae that grew wild in Biskra and El Oued city (Northeastern Algerian Sahara). This study focused on obtaining isolates of legume nodule bacteria (LNB) from the plant G. saharae and evaluated their effectiveness in forming a symbiotic relationship with the legume species Vigna unguiculata through cross-inoculation. Additionally, the study aimed to identify the successful cross-inoculation group of LNB strains based on their phenotypic characteristics. The growth capacity of isolates under varying salinity conditions [NaCl] and pH levels was investigated using a spectrophotometer (96-microplate reader). The API 20NE and API 20E systems were used to identify the biochemical characteristics of the isolates. In addition, the rhizospheric soil samples from the two study sites were analyzed using standard analytical techniques of soil. All isolates established effective symbioses with Vigna unguiculata, were Gram-negative rods, and were fast-growing. The optimal growth temperature was between 28 °C and 37 °C; some isolates were thermophiles and specifically withstood extreme heat between 45-50 °C. Furthermore, they demonstrated a wide tolerance range to pH (5–10) with salt tolerance ranging from 100 mM to 500 mM. Biochemical results revealed that the isolates assimilated various sources of carbon and nitrogen and displayed numerous enzyme activities. Physicochemical analysis revealed that all the soils were deficient in nutrients and had an alkaline pH. This study enabled us to identify the effective stress-tolerant strains, which could be used in the future to inoculate plants for environmental applications. Keywords: Rhizobia, morpho-physiological and biochemical characterization.

Molecular Patterns and Antimicrobial Resistance Characterization of Salmonella enterica Non-Typhoidal from Human, Food, and Environment Samples Isolated in Luanda, Angola

The aim of this study was to characterize the antimicrobial resistance phenotype and genotype of non-typhoidal Salmonella spp. isolated in Luanda, Angola. Between 2013 and 2015, human clinical samples, food, and environmental samples (n = 290) were collected at different regions of Luanda city and screened for the presence of Salmonella spp. Bacterial isolates were preliminarily identified using the API 20E Kit, and their identification was confirmed using PCR and serotyping. All Salmonella spp. isolates were tested by minimum inhibitory concentration against 19 antimicrobials. The isolates were also screened using PCR for the presence of resistance genes (blaOXA-1, blaSHV, blaTEM, sul1, sul2, sul3, qnrA, qnrB, qnrS, qnrC, qnrD, aac(6′)-Ib, dfrIa [targeting dfrA1, dfrA5, dfrA15, dfrA15b, dfrA16, dfrA16b] and dfrA12, cmlA, and floR) and typed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Salmonella enterica non-typhoidal was detected in 21.3% of the clinical samples (n = 32/150), 11.1% of the food samples (n = 10/90), and 26% of the environmental samples (n = 13/50). Serotyping revealed that the monophasic variant of Salmonella Typhimurium (Salmonella enterica serovar 4,[5],12:i:-) was detected in 38.1% of the samples. Moreover, serovar Salmonella Enteritidis was the second most frequent. Only 7.3% of the isolates were resistant to at least one antimicrobial. Furthermore, isolates from different origins (clinical, environmental, and food) were associated with the same lineages, Salmonella Enteritidis ST11 and S. enterica ser. Typhimurium ST313. The detection of S. enterica serovar 4,[5],12:i:- in different settings reinforces the need for a One Health approach to control this zoonosis in Angola.

Prevalence of Bacterial Urinary Tract Infection among Pregnant Women in Wasit Province in Iraq

Background: Urinary tract infections (UTIs) are the most common type of infection during pregnancy. UTI include a spectrum of disorders, ranging from those affecting the lower urinary tract, like asymptomatic bacteriuria and cystitis, to those affecting the kidney, such as pyelonephritis. In pregnancy UTI can lead to preterm labor, anemia and septicemia. Escherichia coli is the most common pathogenic microorganisms associated with UTI and counting up to 60–80% of all UTIs. Objective: The aim of this study is to determine the prevalence of urinary tract infection (UTI) among pregnant women in wasit province. Materials and Methods: Mid-stream urine samples from 100 pregnant women who attending to Al- Shaheed Fairuz hospital and private clinics in wasit province between the periods of March 2024 to July 2024 with the age range of (16-35) years. The urine samples were cultured on MacConkey and blood agars then examined by Macroscopic and Microscopic examination by Gram's stain. The colonies were examined by biochemical tests (Oxidase, catalase, manitol salt agar, novobiocin tests, IMViC) and confirmatory tests (API 20E and API Staph) for isolates. Results: The results showed that the percentage of bacterial growth culture among the pregnant women was 33%. Gram negative bacteria were the higher percentage with (73%). Eschirishia coli bacteria were the most frequently isolated organism (40%) while, the lower percentage for Streptococcus spp. (3%). UTI in pregnant women was commonest in the age group 20-24 years (42%). According to trimesters of pregnancy, the highest percentage of UTI was in third trimester (43%) followed by second trimester (39%), while the the first trimest was the the lowest percentage with (18%). Conclusions: Urinary tract infections (UTIs) are common during pregnancy. Untreated UTIs can lead to complications such as preterm labor, low birth weight and sepsis. E.coli is the predominant pathogen presently causing urinary tract infections at all stages of pregnancy. Urine culture and sensitivity tests are mandatory for all pregnant women during the different trimesters.

Mass Mortality of the Invasive Sea Urchin Diadema setosum in Türkiye, Eastern Mediterranean Possibly Reveals Vibrio Bacteria Infection

ABSTRACTThe invasive Diadema setosum (Leske, 1778) long‐spined sea urchin has been in the Mediterranean Sea since 2006, and then, it has been known that its population density has reached high values on the southern Aegean coasts of Türkiye. This study aimed to report the mass mortality event of D. setosum with the determined agent on the Aegean coast of Türkiye. In addition, it was targeted to provide information on the current status of its density based on seasonal SCUBA diving observations for the period between January 2023 and November 2023. The results showed that the mass mortality of D. setosum was determined in Muğla, Aegean Sea, in August 2023. Spine loss, the mucoid layer at the bottom of the appendages, and the outer body surface were observed as the clinical symptoms of the diseased samples. Bacterial growth was detected on Vibrio‐selected TCBS agar plates, while no other parasitic agents were determined in the coelomic fluid of sea urchins. The results of the biochemical (API 20E) and molecular tests confirmed the isolated bacteria as Vibrio spp. The density of D. setosum showed statistically significant temporal changes, and the highest and lowest mean density values were recorded in autumn and spring, respectively. Environmental stressors, such as increasing sea temperature levels, affect the marine ecosystem and lead to opportunistic pathogens. Long‐term monitoring of disease outbreaks is necessary to understand the interactions between species and the ecosystem.

Investigation of vibriosis caused by Vibrio anguillarum in rainbow trout (Oncorhynchus mykiss).

Identifying pathogenic microorganisms causing disease is important for epidemiological research, antimicrobial therapy, and control. The current study was carried out to use different methods for the identification of Vibrio anguillarum from diseased rainbow trout (Oncorhynchus mykiss) obtained from Türkiye (Muğla-Fethiye), the damage caused by the pathogenic microorganism in the tissues and organs, and the determination of the antibiotic effective against the pathogen. Hemorrhagic and ulcerative skin lesions and diffuse petechial hemorrhage in the internal organs were clinically detected in diseased fish obtained from the rainbow trout farm. Bacteria isolated from diseased fish were subjected to analysis using conventional bacteriological methods, a commercial bacterial identification test kit (API), an automated bacteria identification system known as Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS), BD Phoenix™, and 16S rRNA sequence analysis. All isolated bacteria were identified as V. anguillarum by API 20E and conventional bacteriological method. These results have been confirmed with 16S rRNA sequence analysis. However, the isolated bacteria were identified as Grimontia hollisae (syn. Vibrio holisae) with the BD Phoenix system. Histologically, tissue damage such as melano-macrophage centers and necrosis in the kidney and spleen, hyperemia and mononuclear cell infiltration in the liver, as well as mononuclear cell infiltration on muscles, talengectiasis in the gill tissue was observed. In addition, it has been determined that the most effective antibiotic against the pathogen was enrofloxacin. When comparing all identification methods used for this pathogen causing tissue damage, it was demonstrated that the MALDI-TOF MS provides better results than other methods in terms of cost and identification time, and it could be used as an alternative to the conventional method to the identification of V. anguillarum.

Microbiological identification of some culturable native strains associated with the digestive tract in Panaque cochliodon

Objective. To characterize by microbiology the native strains associated to the intestinal tract of the species Panaque cochliodon. Materials and methods. Three adult specimens captured in the Magdalena River that were transported and sacrificed under animal welfare regulations were used. Dissection of the intestinal tract was performed, obtaining samples for microbiological isolation, using culture media, purifying the microorganisms, performing metabolic biochemical tests and API 20E (Biomeriux) tests for their identification. Results. Information on the microbial population structure was obtained, reporting the phylum Proteobacteria with the species: Pantoea sp, Erwinia sp, Providencia stuarti, Providencia alcalifaciens, Serratia ficaria, Citrobacter koseri and the phylum Firmicutes with the species: Bacillus sphaericus, Bacillus subtilis, Bacillus thurigiensis, Bacillus mycoides, Bacillus coagulans and Bacillus circulans. Conclusion. The predominant culturable microorganisms in P. cohliodon belong to the phylum Proteobacteria and the phylum Firmicutes.

Exploring the Resistance Pattern of Salmonella Typhoidal Species

Background: Enteric fever/ typhoid, an infection caused by Salmonella enterica serovars: typhi and paratyphi, is continuously increasing with the emergence of antibiotic resistant Salmonella. According to WHO, an estimated 110,000 people die from it every year. Objective: The primary objective of this study was to investigate the antibiotic susceptibility pattern of typhoidal Salmonella species in blood cultures of patients. Methodology: This was a cross-sectional study, executed at Microbiology Laboratory (Diagnostic Centre), PAF Hospital Faisal Base Karachi, from August 2023 to March 2024. Salmonella typhi in all clinically suspected cases of typhoid fever was identified through conventional techniques following standard microbiological protocols. Assessment of antibiotic susceptibility was performed by Kirby-Bauer disk diffusion method. Results: Out of 1178 blood samples culture 54 were gram positive bacteria, 193 were gram negative bacteria and 931 samples showed no growth. Among the gram-negative sample, 140 tested positive for Salmonella enterica typhoidal species. API 20E was used to further differentiate and identify the Salmonella species, 131 cases were Salmonella typhi and 9 paratyphi. Out of 140 Salmonella isolates, 77% (108) were resistant to ampicillin, 49.2% (69) to Ceftriaxone, 35% (49) to Sulfamethoxazole, and 47% (66) to Ciprofloxacin. Azithromycin and Meropenem remained sensitive in all 140 isolates of Salmonella typhoidal species. Conclusion: This study identified the susceptibility profiles of Salmonella typhoidal species. Our findings underscore the importance of continuous surveillance and judious use of antibiotics to combat the emergence of antibiotic –resistant strains. Further research is warranted to continuously monitor and adapt treatment protocols in response to evolving resistance in Salmonella typhoidal species.

تثبيط تكوين الاغشية الحيوية في بكتيريا Agrobacterium tumefaciens بالمادة الطافية الخالية من خلايا Pseudomonas aeruginosa والمحللة بجهاز GC-MS

تعد Agrobacterium tumefaciens من أنواع البكتيريا المسببة للأمراض النباتية المهمة اقتصاديا، وتحصل الاصابة نتيجة استخدام اغشيتها الحيوية لالصاق نفسها بالجروح المتكونة على سطح العائل النباتي، ونظرًا لمحدودية العلاجات الحالية وفعاليتها، أصبح البحث عن عوامل جديدة مضادة لهذه البكتيريا أمرا ضروريا، لذلك تم تسليط الضوء في هذه الدراسة على معرفة التأثير التثبيطي للمادة الطافية الخالية من خلايا Pseudomonas aeruginosa على تكوين الاغشية الحيوية من قبل A. tumefaciens، فضلا عن تحديد المركبات الكيمياوية المكونة لها بجهاز GC-MS. بناءا على ذلك عزلت P. aeruginosa من التربة وتم تشخيصها باستخدام العدةAPI 20 E وتفاعل البلمرة المتسلسل باستخدام الجين 16S rRNA واظهرت تطابقا بنسبة 93% مع البكتيريا القياسية Pseudomonas sp.SeaQual P_B_845W, MT626817.1 في بنك الجينات. وعند الكشف عن قدرة المادة الطافية المجفدة وبتسعة تراكيز10، 15، 20 ،25 ، 30، 35، 40، 45، 17، 17، 15 ،على التوالي. وحسب عدد القمم التي تم إنتاجها بعد حقنها بجهاز GC، تبين انها مكونة من 30 مادة كيميائية، وبعد إدخال هذه المعلومات في جهاز MS، تم تشخيصها بأسمائها ومنها(Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-، Hexadecanoic acid, methyl ester، Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl)-،9-Octadecenoic acid (Z)-,methyl ester،cis-13-Octadecenoic acid,methyl ester،Octadecanoic acid, methyl ester)، البالغ وزنها الجزيئي(154، 270، 210، 296، 296، 298) دالتون ومقدار المساحة التي تشغلها (9.38، 19.12، 6.8، 4.45، 8.33، 5.90)%، على التوالي. أهم النتائج التي شخصتها هذه الدراسة هو تحديد المركبات الكيميائية لأول مرة للمادة الطافية الخالية من خلايا الزائفة الزنجارية وتأثيرها المثبط على إنتاج الأغشية الحيوية من قبل A. tumefaciens.

Las Bolitas Syndrome in Penaeus vannamei Hatcheries in Latin America.

In September 2023, several hatcheries in Latin America experienced significant mortality rates, up to 90%, in zoea stage 2 of Penaeus vannamei. Observations of fresh mounts revealed structures resembling lipid droplets, similar to those seen in a condition known as "las bolitas syndrome". Routine histopathological examinations identified detached cells and tissues in the digestive tracts of affected zoea, contrasting with the typical algal cell contents seen in healthy zoea. Polymerase chain reaction (PCR) testing for over 20 known shrimp pathogens indicated minimal differences between diseased and healthy batches. Both groups tested negative for acute hepatopancreatic necrosis disease (AHPND) but positive for Vibrio species and Rickettsia-like bacteria in the diseased samples. Histological analyses of the affected zoea revealed characteristic tissue degeneration in the hepatopancreas, forming spheres that eventually migrated into the upper gut, midgut, and midgut caeca, a pathology identified as bolitas syndrome (BS). Microbiological assessments revealed Vibrio species at concentrations of 106 CFU zoea/g in affected zoea, approximately two orders of magnitude higher than in healthy zoea. Bacterial isolation from both healthy and BS-affected zoea on thiosulphate-citrate-bile salts-sucrose (TCBS) agar and CHROMagar™ (Paris, France), followed by identification using API 20E, identified six strains of Vibrio alginolyticus. Despite similarities to "las bolitas syndrome" in fresh mounts, distinct histopathological differences were noted, particularly the presence of sloughed cells in the intestines and variations in hepatopancreatic lobes. This study highlights the critical need for further research to fully understand the etiology and pathology of bolitas syndrome in zoea stage 2 of P. vannamei to develop effective mitigation strategies for hatchery operations.

Parasporins of Bacillus thuringiensis Strain Exhibit Apoptosis-Mediated Selective Cytotoxicity to MDA-MB-231 Cells through Oxidative Stress

Historically, the most important source of both antibiotics and anticancer medications has been microorganisms. Bacillus thuringiensis (Bt) is one of the most prominent bacterial species used as a therapeutic agent targeting cancerous cells in recent worldwide investigations. This study was designed to isolate, molecularly identify, and discover novel Saudi Arabian Bt strains that selectively exhibit cytotoxic properties against MDA-MB-231, a human triple-negative breast cancer (TNBC) cell model. The bacterial strain under investigation was biochemically typed using API 20E and API CH50 and molecularly typed using 16S rDNA sequencing. Flow cytometry and immunoblotting were performed to elucidate the mechanism-of-action (MOA). Molecular typing confirmed the identity of the isolated non-hemolytic strain to be Bt and was named Bt HAU-145. Microscopic examination showed that the strain possessed a parasporal (PS) crystal protein with a spherical morphology. Data of cytotoxicity assay based on MTT revealed that Bt HAU-145 strain exhibited selective and potent cytotoxicity against MDA-MB-231, with a 50 percent inhibition (IC50) of 28 µg/ml. FACS analysis revealed that PS proteins induced both late and early apoptosis in a ROS-dependent manner. Immunoblotting assays showed increased expression of caspase-3 in response to PS treatment, paralleled by a reduction in Bcl-2 expression. This is the first study to investigate the MOA of PS proteins from the Saudi Arabian Bt strain, showing an induction of apoptosis through a ROS-dependent mechanism in TNBC cells. It is hoped that PS-based therapeutic strategies will be investigated at the preclinical scale in non-human primates prior to the clinical scale in randomized clinical trials.

Diversity and antimicrobial susceptibility patterns of clinical and environmental Salmonella enterica serovars in Western Saudi Arabia.

The diverse environmental distribution of Salmonella makes it a global source of human gastrointestinal infections. This study aimed to detect Salmonella spp. and explore their diversity and antimicrobial susceptibility patterns in clinical and environmental samples. Pre-enrichment, selective enrichment, and selective plating techniques were adopted for the Salmonella detection whereas the API 20E test and Vitek Compact 2 system were used to confirm the identity of isolates. Salmonella serovars were subjected to molecular confirmation by 16S rDNA gene sequencing. Disc diffusion method and Vitek 2 Compact system determined the antibiotic susceptibility of Salmonella serovars. Multiple antibiotic resistance index (MARI) was calculated to explore whether Salmonella serovars originate from areas with heavy antibiotic usage. Results depicted low Salmonella prevalence in clinical and environmental samples (3.5%). The main detected serovars included Salmonella Typhimurium, S. enteritidis, S. Infantis, S. Newlands, S. Heidelberg, S. Indian, S. Reading, and S. paratyphi C. All the detected Salmonella serovars (27) exhibited multidrug resistance to three or more antimicrobial classes. The study concludes that the overall Salmonella serovars prevalence was found to be low in environmental and clinical samples of Western Saudi Arabia (Makkah and Jeddah). However, antimicrobial susceptibility patterns of human and environmental Salmonella serovars revealed that all isolates exhibited multidrug-resistance (MDR) patterns to frequently used antibiotics, which might reflect antibiotic overuse in clinical and veterinary medicine. It would be suitable to apply and enforce rules and regulations from the One Health approach, which aim to prevent antibiotic resistance infections, enhance food safety, and improve human and animal health, given that all Salmonella spp. detected in this investigation were exhibiting MDR patterns.

Diagnostic accuracy of automation and non-automation techniques for identifying Burkholderia pseudomallei: A systematic review and meta-analysis

BackgroundBurkholderia pseudomallei, a Gram-negative pathogen, causes melioidosis. Although various clinical laboratory identification methods exist, culture-based techniques lack comprehensive evaluation. Thus, this systematic review and meta-analysis aimed to assess the diagnostic accuracy of culture-based automation and non-automation methods. MethodsData were collected via PubMed/MEDLINE, EMBASE, and Scopus using specific search strategies. Selected studies underwent bias assessment using QUADAS-2. Sensitivity and specificity were computed, generating pooled estimates. Heterogeneity was assessed using I2 statistics. ResultsThe review encompassed 20 studies with 2988 B. pseudomallei samples and 753 non-B. pseudomallei samples. Automation-based methods, particularly with updating databases, exhibited high pooled sensitivity (82.79%; 95% CI 64.44–95.85%) and specificity (99.94%; 95% CI 98.93–100.00%). Subgroup analysis highlighted superior sensitivity for updating-database automation (96.42%, 95% CI 90.01–99.87%) compared to non-updating (3.31%, 95% CI 0.00–10.28%), while specificity remained high at 99.94% (95% CI 98.93–100%). Non-automation methods displayed varying sensitivity and specificity. In-house latex agglutination demonstrated the highest sensitivity (100%; 95% CI 98.49–100%), followed by commercial latex agglutination (99.24%; 95% CI 96.64–100%). However, API 20E had the lowest sensitivity (19.42%; 95% CI 12.94–28.10%). Overall, non-automation tools showed sensitivity of 88.34% (95% CI 77.30–96.25%) and specificity of 90.76% (95% CI 78.45–98.57%). ConclusionThe study underscores automation's crucial role in accurately identifying B. pseudomallei, supporting evidence-based melioidosis management decisions. Automation technologies, especially those with updating databases, provide reliable and efficient identification.

Analysis of Microbiological Hazards in Turmeric Tamarind Traditional Drink Products

Turmeric tamarind is a traditional drink with a sour and sweet taste traditionally made on a household scale. This drink is popular among the community, students, and local tourists from various regions visiting Yogyakarta. Due to its relatively simple production process and potentially suboptimal hygiene standards, the risk of bacterial contamination remains a concern for public health. Therefore, this study aimed to determine the presence or absence of bacterial contaminants, namely Escherichia coli, Salmonella sp and Staphylococcus aureus according to SNI 2019 standards. Samples were obtained from 11 locations of turmeric tamarind drink sales and inoculated into Chromocult Coliform Agar (CCA) differential selective medium, Salmonella Sigella Agar (SSA), and Baird Parker Agar (BPA) for E. coli isolation, Salmonella sp, and S. aureus, respectively. Subsequently, colonies were prified and screened through biochemical tests such as IMViC, Urease test, and sugar fermentation test for lactose, maltose, and glucose. Suspected isolates were also confirmed using API 20E and API STAPH tests. The results showed that E. coli, Salmonella sp, and S. aureus were not reported while Klebsiella pneumoniae ssp pneumoniae 1, Citrobacter freundii, Staphylococcus xylosus, and Staphylococcus lentus were obtained.

Apirhabdus apintestini gen. nov., sp. nov., a member of a novel genus of the family Enterobacteriaceae, isolated from the gut of the western honey bee Apis mellifera.

A Gram-negative, motile, rod-shaped bacterial strain, CA-0114T, was isolated from the midgut of a western honey bee, Apis mellifera. The isolate exhibited ≤96.43 % 16S rRNA gene sequence identity (1540 bp) to members of the families Enterobacteriaceae and Erwiniaceae. Phylogenetic trees based on genome blast distance phylogeny and concatenated protein sequences encoded by conserved genes atpD, fusA, gyrB, infB, leuS, pyrG and rpoB separated the isolate from other genera forming a distinct lineage in the Enterobacteriaceae. In both trees, the closest relatives were Tenebrionicola larvae YMB-R21T and Tenebrionibacter intestinalis BIT-L3T, which were isolated previously from Tenebrio molitor L., a plastic-eating mealworm. Digital DNA-DNA hybridization, orthologous average nucleotide identity and average amino acid identity values between strain CA-0114T and the closest related members within the Enterobacteriaceae were ≤23.1, 75.45 and 76.04 %, respectively. The complete genome of strain CA-0114T was 4 451669 bp with a G+C content of 52.12 mol%. Notably, the apparent inability of strain CA-0114T to ferment d-glucose, inositol and l-rhamnose in the API 20E system is unique among closely related members of the Enterobacteriaceae. Based on the results obtained through genotypic and phenotypic analysis, we propose that strain CA-0114T represents a novel species and genus within the family Enterobacteriaceae, for which we propose the name Apirhabdus apintestini gen. nov., sp. nov. (type strain CA-0114T=ATCC TSD-396T=DSM 116385T).

Detection of carbapenemase producing enterobacterales using the Modified Hodge Test from clinical isolates in Colombo South Teaching Hospital and Sri Jayewardenepura General Hospital, Sri Lanka in 2017

Introduction: Enterobacterales is a large family of Gram-negative bacilli including many pathogens. Carbapenemase producing Enterobacterales (CPEs) have emerged as a global threat. This study was conducted to detect carbapenemase production by Enterobacterales isolates from clinical specimens and to correlate the occurrence of CPE with age, gender, and duration of hospital stay of the patients included in the study.Methods: A descriptive cross-sectional study was carried out using 120 consecutive, non-repetitive clinical isolates identified as Enterobacterales by the microbiology laboratories of Colombo South Teaching Hospital and Sri Jayewardenepura General Hospital. The demographic data of the patients was gathered and used to analyse the correlation with CPE occurrence. All isolates were identified up to species level using API 20E kits. Screening for detection of carbapenemase was carried out using meropenem, imipenem, and ertapenem disks. The Modified Hodge Test (MHT), which is one of the suggested procedures to determine carbapenemase production, was performed to identify carbapenemase producing isolates.Results: Of the120 isolates, 14 (11.7%) were resistant to at least one of the carbapenems tested. MHT detected carbapenemase production in ten (8.3%) isolates. The majority of these isolates were Klebsiella sp. (6; 60.0%). Of carbapenemase producing isolates, six were from urine specimens (60%). Eight (80.0%) of the CPE harbouring patients were males and eight (80.0%) were aged above 50 years. The mean duration of hospital stay of the patients was 7.2 days (±SD 5.65days).Conclusion: The proportion of CPE was 8.3% according to this study and emphasises the importance of conducting more studies on the prevalence of CPE in Sri Lanka.

Prevalence of Extensively and Pan-drug-resistant Acinetobacter spp. in Nosocomial Infections in Western Saudi Arabia

Multidrug resistance patterns of Acinetobacter spp. have led to their emergence as an important source of nosocomial infections. This study investigated the prevalence and clinical characteristics of Acinetobacter spp. in hospital-acquired wound and urinary tract infections. A total of 432 samples [wound swabs (210) and urine samples (222)] were analyzed for the presence of Acinetobacter spp. through selective culturing on MacConkey and Leeds Acinetobacter medium followed by identification with API 20E strips and Vitek 2 compact system. Antimicrobial susceptibility was assessed by adopting the disk diffusion method on Muller-Hinton agar, whereas the minimum inhibitory concentration procedure was carried out by using a ComASP™ Colistin test kit. Biofilm formation was examined using microtiter plates and following the crystal violet staining method. PCR was performed to amplify virulence (lasB, bap, and plcN) and antimicrobial resistance (blaOXA-51like) genes. The results revealed a low prevalence of Acinetobacter spp. (1.85 %) where Acinetobacter baumannii was the predominant species. Acinetobacter baumannii isolates harbored blaOXA-51-like gene to exert extensive or pan-drug resistance. Most Acinetobacter baumannii isolates demonstrated weaker to moderate biofilm-forming capabilities and carried the bap gene. Acinetobacter baumannii isolates lacked the combination of virulence factors encoding lasB and plcN genes. Acinetobacter baumannii infections are rising in Saudi Arabia. The results of this study highlight the epidemiology of virulent and antibiotic-resistant Acinetobacter spp., particularly A. baumannii, in Saudi Arabia. The detailed elaboration on the diversity, virulence, and antimicrobial susceptibility patterns of Acinetobacter spp. in Saudi Arabia requires further in-depth molecular investigations.

Isolation and identification of bacterial pathogens causing ice-ice disease in Eucheuma cottonii seaweed at Seira Island Waters, Tanimbar Islands District, Maluku, Indonesia

Abstract. Tuhumury NC, Sahetapy JMF, Matakupan J. 2024. Isolation and identification of bacterial pathogens causing ice-ice disease in Eucheuma cottonii seaweed at Seira Island Waters, Tanimbar Islands District, Maluku, Indonesia. Biodiversitas 25: 964-970. Seaweed cultivation has proven to be beneficial and enhances the well-being of communities. Production of seaweed in the waters of Maluku has increased despite facing the problem of ice-ice disease. Therefore, this research aimed to isolate and identify bacteria on the Eucheuma cottonii seaweed species affected by ice-ice disease in the waters of Seira Island, Tanimbar Islands District, Maluku. It was conducted in August 2023 at three observation stations, and water quality samples were collected in situ at all three stations, including temperature, salinity, pH, dissolved oxygen (DO), and turbidity. Furthermore, bacterial isolation and identification were carried out at the Laboratory of Marine Aquaculture, Ambon. Bacteria isolation and purification were performed on TCBS media using the streak plate technique, while identification used the API 20E test. The results showed that water quality parameters supported seaweed growth. Four bacteria that caused ice-ice disease in the waters of Seira were identified as Vibrio alginolyticus, Vibrio fluvalis, Vibrio cholerae, and Aeromonas caviae. Moreover, all four bacteria found were gram-negative and pathogenic. Bacteria belonging to the genera Vibrio and Aeromonas have been proven to cause infections in marine organisms, including seaweed. In conclusion, four types of bacteria were identified in seaweed affected by ice-ice disease.

Study the Phenotypic and Antimicrobial Susceptibility of Pseudomonas aeruginosa Isolated Clinically from Baghdad Hospitals

Pseudomonas aeruginosa, is negative to gram stain, take the rod shape and considered as strictly aerobic. the most affect bacteria in nosocomial infection. P.aeruginosa is an opportunistic pathogen bacterium that can made serious infections in human who are immune compromised, such as urinary tract infection, skin and ear infection, and others. One hundred twenty one sample were taken from patients in different sites of infections (ear, wound, UTI , burn) from AL-Yarmouk, Ghazi Al-Hariri hospitals in Baghdad during the period from November 2016 to February 2017. Fifty seven isolates were diagnoses as P. aeruginosa by characteristic in culture media, tests and API 20E. forty isolates involved in our study ten from each source. Antibiotic susceptibility test performed for forty clinically isolates of P. aeruginosa by the disk diffusion method against some antibiotics belong to different group and the results revealed that bacteria is multi-drug resistance (MRD) as well, revealed the most compound that have activity against Pseudomonas. aeruginosa were Imipenem, Pipracillin, and Ceftazidime. Biofilms are currently estimated and the P. aeruginosa reflect high ability to produce biofilm. All isolates used in this study formed biofilm with differences in thickness of formed layer. In this study, we concluded P. aeruginosa is one of the most common gram-negative bacteria involved in hospital infections causing opportunistic infection because they have intrinsically and acquired resistant to a number of antimicrobial agents and produces a number of exoproducts which are implicated in the pathogenesis of P. aeruginosa infections.

Analysis of Plasmid-Mediated Quinolone Resistance (PMQR) Genes Among Escherichia coli Strains Isolated from Patients with Urinary Tract Infection in Eastern Turkey

Aim: Urinary tract infections are a common occurrence worldwide, with females being more susceptible than males. Infections can manifest as asymptomatic bacteriuria or more severe forms such as sepsis. This study aims to analyse the plasmid-mediated quinolone resistance genes of Escherichia coli (E. coli) responsible for causing urinary tract infections.
 Material and Method: Microbiological evaluation was conducted using conventional culture method, API 20E (API-bioMérieux, France), antibiotic susceptibility tests, and polymerase chain reaction (PCR).
 Results: During the study period, 145 E. coli strains were collected from 96 (66.2%) outpatients and 49 (33.8%) hospitalised patients with urinary tract infections (UTI). The resistance rates of E. coli strains to quinolone antibiotics were as follows: nalidixic acid in 102 strains (70.3%), ciprofloxacin in 96 strains (66.2%), ofloxacin in 73 strains (50.3%), gatifloxacin in 47 strains (32.4%), levofloxacin in 41 strains (28.3%) and moxifloxacin in 32 (22%) strains. Based on the PCR test results, it was found that 80% (116) of the isolates carried at least one PMQR gene. Furthermore, 31.9% (37/116) of the isolates were found to carry two or three PMQR markers simultaneously.
 Discussion and Conclusion: The objective of this study was to establish the prevalence of quinolone resistance among E. coli strains isolated from patients with UTI. To control the high rate of spread of antibiotic-resistant microorganisms, it is crucial for all healthcare facilities to implement more careful management of antibiotic use and adhere to procedures set by infection control programmes.

Distribution of antibiotic resistant bacteria and genes in sewage and surrounding environment of Tórshavn, Faroe Islands

Several studies have investigated the effects of swimming in sewage-polluted recreational beach water, highlighting the associated health hazards. To mitigate potential pathogen transmission, it is imperative that the polluted water is released away from recreational waters and foreshores, where children tend to play. At present, domestic sewage in the Faroe Islands solely undergoes primary wastewater treatment within primary settling tanks before being discharged into the ocean. Effluents are a major anthropogenic source of antibiotic resistance genes and antibiotic resistant bacteria, which are released into the environment. The aim of this study was to investigate antibiotic resistant Gram-negative bacteria and antibiotic resistance genes in influents and effluents of wastewater subjected solely to primary treatment, along with their release into the environment during both summer and winter. Water samples were collected from influents and effluents as well as with increasing distance away from the wastewater outlet and from nearby tidepools. Samples were cultured on MacConkey agar with four different antibiotics for detection of antibiotic-resistant bacteria and antibiotic resistance genes were quantified by droplet digital PCR. All multi-drug resistant bacteria were identified using the API 20E kit. We observed an overall decrease of the abundance of Gram-negative bacteria from the effluents compared to influents, however, we observed the opposite trend in the antibiotic resistance genes. Antibiotic resistant bacteria and antibiotic resistance genes in addition to multi-drug resistant bacteria were found in the surrounding oceanic and several terrestrial tidepool samples. Of the multi-drug resistant bacteria, we found, e.g., Escherichia coli, P. aeruginosa, and A. hydrophila species, which can be pathogenic, potentially causing an infection if encountering a host. These results indicate a relatively wide pollution range of the effluents from the septic tank and treated sewage released into the environment, posing a potential hazard for both humans and wildlife.