Oxidase-positive Research Articles

Occurrence of Staphylococcus aureus, Staphylococcus epidermidis, and Staphylococcus pseudintermedius colonization among veterinarians in the province of Malaga, Spain

Background and Aim: Staphylococcus pseudintermedius and Staphylococcus aureus are common colonizing pathogens in companion animals. These opportunistic pathogens can cause infections of varying frequency and severity in humans and pets. Studies on Staphylococcus colonization in veterinarians are scarce. This study aimed to investigate the colonization of the nostrils and hands by S. aureus, Staphylococcus epidermidis, and S. pseudintermedius among healthy clinical practice veterinarians in the province of Malaga (Spain), with a particular focus on their potential antibiotic resistance. Materials and Methods: A request for voluntary participation was extended to professionals from the Official College of Veterinarians of Malaga. Nasal and hand swabs were collected by two trained technicians in January 2024, and all samples were delivered to the laboratory within 24 h. Gram staining, catalase, oxidase, and coagulase tests were performed. The susceptibility of the isolated bacteria to 11 antibiotics was evaluated. Results: A total of 50 clinical practice veterinarians were enrolled in the study, comprising 36 women and 14 men from 31 veterinary clinics across Málaga province. A total of 32% of the nasal samples yielded S. aureus, whereas 64% were found to contain S. epidermidis. In total, 30% of the hand samples yielded S. aureus and 30% yielded S. epidermidis. The participants did not exhibit any strains of S. pseudintermedius in their nasal samples or hands. Two strains (11.1%) of methicillin-resistant S. aureus were isolated from 18 strains isolated from nostrils. Furthermore, a high prevalence of S. aureus strains resistant to ampicillin (94.4%) and amoxicillin (72.2%) was observed. Conclusion: The colonization profiles of veterinary professionals were similar to those observed in the general population. Further research is required among veterinary professionals, companion animals, and their owners to better understand the colonization processes and the pet-human interface within a “One Health” approach. Keywords: antibiotic resistance, drug-resistant Staphylococcus aureus, one health, Seroprevalence, Staphylococcus colonization, veterinarians.

Asymptomatic nasopharyngeal carriage of multidrug resistant bacteria among children at University of Gondar Hospital Northwest Ethiopia Revealing Hidden Health Risks

Gram-negative bacteria in the nasopharynx can eventually invade bacteria-limited sites and cause serious illnesses such as meningitis, otitis media, and pneumonia. However, data related to the carriage of these bacteria in children attending outpatient departments in the study area are limited. To assess nasopharyngeal carriage, antibiotic susceptibility patterns, and associated factors of gram-negative bacteria among children attending the outpatient department at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia. A hospital-based cross-sectional study was conducted from September 1, 2023, to December 30, 2023. A total of 385 children aged 3 to 14 years were enrolled using a systematic random sampling technique. Sociodemographic and clinical data were collected using a semistructured questionnaire. A total of 385 nasopharyngeal samples were collected using a sterile specimen collection nasopharyngeal swab, transported using Amie’s transport medium, and subsequently inoculated on chocolate agar, blood agar, modified thayer martin agar, and MacConkey agar plates. Bacterial species were identified by colony morphology, Gram staining, and biochemical tests such as oxidase tests, satellitism tests, and carbohydrate utilization tests. An antibiotic susceptibility test was performed using the Kirby–Bauer and modified Kirby–Bauer methods on Mueller–Hinton agar plates. The data were entered into Epi-Data version 4.6.0.6 and exported to SPSS version 25 for analysis. The adjusted odds ratio at a 95% confidence interval with a P value of < 0.05 in the binary logistic regression model was considered to indicate statistical significance. The overall nasopharyngeal carriage of gram-negative bacteria was 146 (37.9%) (95% CI: 33.2–42.9). Among these, nonfastidious gram-negative bacteria represented 45 (11.7%), followed by M. catarrhalis 41 (10.6%), N. meningitidis 34 (8.8%), and H. influenzae 26 (6.8%). The isolates exhibited high resistance to tetracycline (85; 75.9%), trimethoprim-sulfamethoxazole (105; 71.9%), ampicillin (76; 67.9%), and amoxicillin/clavulanic acid (60; 69.8%) but high susceptibility to meropenem (122; 83.6%), gentamicin (73; 84.9%), and minocycline (87; 72.5%). There were 99 total multidrug-resistant strains (67.8%, 95% CI: 59.7–75.0). Male sex (AOR = 1.785, 95% CI: 1.102–2.892, P = 0.019), smoking (AOR = 2.675, 95% CI: 1.149–6.230, P = 0.022), and large family size (≥ 5) (AOR = 1.857, 95% CI: 1.140–3.023, P = 0.013) were risk factors for nasopharyngeal colonization. Increased nasopharyngeal colonization of multidrug-resistant gram-negative isolates was observed in this study. Gentamicin, minocycline, and meropenem were the most effective antibiotics for the tested isolates. Bacterial colonization increased with increasing family size, smoking status, and male sex. Therefore, a definitive diagnosis in the outpatient pediatric department should be based on culture and susceptibility test results.

Faucets and keyboards as a major reservoir of gram-negative bacilli in a regional hospital in Northeast Brazil

Infections related to healthcare assistance represent a major public health problem worldwide, and the main etiological agents associated with these infections are microorganisms also found on inanimate hospital surfaces. Therefore, understanding the dynamics of bacterial contamination of hospital surfaces is essential for professionals working in these services. Objective: To evaluate the contamination of inanimate surfaces by Gram-negative bacilli (GNB) in a Regional Hospital in Northeast Brazil. Methods: This is an observational and cross-sectional study. Samples were collected from different hospital wards using a sterile swab soaked in saline solution. They were then plated on MacConkey agar and incubated at 36°C for 48 hours. The bacterial isolates were counted and identified using the Gram staining method, TSI agar, and oxidase test. Results: Out of the 150 samples collected, 71 showed no microbiological growth, 11 showed growth of filamentous fungi, and 66 showed growth of Colony Forming Units (CFU). A total of 132 morphologically distinct colonies were identified, with 55 identified as yeasts and 77 as GNB. In the evaluation of hospital wards, the Adult Emergency Room and the Orthopedic Surgical Clinic showed the highest growth of GNB. Regarding the surfaces studied, the most contaminated were faucets, computer keyboards, mattresses, and beds. Conclusions: Hospital areas with higher human traffic are the most contaminated, particularly on frequently handled objects. Therefore, it is necessary to focus on disinfecting these surfaces to break the transmission chain of these etiological agents.

Prevalence and identification of Bacillus species in cellophane, foil, and disposable plastic containers used for food packaging

Background: Foil, cellophane, and disposable plastic containers are commonly employed in food packaging, namely as coatings on food items. Nevertheless, the existence of microbes on the surface and inside the internal composition of these materials gives rise to concerns, as they have the capacity to multiply under ideal circumstances and spread to food items, thereby jeopardizing the safety and quality of the food. Objective: This study aimed at examining the number of microorganisms and identifying specific bacterial species within frequently used food packaging materials like foil, cellophane, and disposable plastic containers. Methods: samples of foil, cellophane, and disposable plastic containers were collected from various popular brands in Iran. The overall bacterial count wضas determined using defibering, flooding, and smear techniques followed by culturing on Tryptone Glucose Extract Agar (TGEA) medium. To identify the bacterial species, various biochemical tests were performed, including fermentation, motility testing, catalase test, oxidase test, and methyl red-Voges Proskauer (MR-VP) test. Results: Bacterial quantity ranged from 0 CFU/g to 8.3 × 103 CFU/g in analyzed samples. All the samples had primary contaminants mainly Bacillaceae family bacteria that could produce spores. Disposable plastic containers had the lowest bacterial count whereas cellophane showed the highest bacterial contamination level. Among Bacillaceae family members Bacillus licheniformis was dominant. Conclusion: The study's findings emphasize the possibility of microbial contamination in food packaging materials, including cellophane. These results indicate that the food packaging industry should adopt a rigorous approach to hazard evaluation and Important Control Points (HACCP) in order to guarantee the microbiological safety and efficacy of packaging materials.

Signaling Molecules in Pseudomonas aeruginosa Response to Antibiotics at Sub-Inhibitory Concentrations

Signaling Molecules are N-acylhomoserine lactones (AHLs) that form the Pseudomonas aeruginosa cell information system which determines gene expressions in a population dependent manner called quorum sensing (QS). Signal molecules, which are chemically varied, control genes expressions to antibiotics resistance, pathogenicity, motility, biofilm development, bioluminescence, secondary metabolite production, and plasmid transfer. This research was aimed to identify the signaling molecules in Pseudomonas aeruginosa response to antibiotics at sub-inhibitory concentrations. One hundred and fifty (150) clinical swab specimens were collected from urinary catheter wound and ear infection of patients and the swabs inoculated using standard microbiology method. The isolates were characterized based on the bacteriological methods such as morphology and biochemical tests. The isolates were further confirmed by species specific PCR by amplification of 16S rRNA and the amplicons were analyzed by gel electrophoresis; and further genomic sequencing was done and blast with NCBI database mining. Disc diffusion method was applied for the antibiotics susceptibility pattern. The isolates cell suspensions were analyzed by Gas Chromatography-Mass Spectrometry (GC-MS). The result of the isolation showed 22(59.46%) from wound infections, 12(32.43%) from ear infections and 3(8.11%) from urinary catheter. The isolates were Gram negative, produced β-hemolysis on blood agar and the morphology is small pigmented circular in form. The isolates showed positive result to catalase, oxidase, citrate, nitrate and indole tests. The amplification of 16S rRNA gene region resulted in the band size of 1500bp PCR product and the BLAST analysis gave 99% similarity. The resistant antibiotics susceptibility showed that Pseudomonas aeruginosa is multidrug resistant bacteria. The GC-MS results obtained from urinary catheter isolates showed four (4) signaling molecules such as N-Octanoyl-L-homoserine Lactone, N-Decanoyl-DL-Homoserine Lactone, N-Dodecanoyl-DL-Homoserine Lactone and N-Tetradecanoyl-L-Homoserine Lactone. Three (3) signaling molecules such as N-Octanoyl-L-homoserine Lactone, N-Decanoyl-DL-Homoserine Lactone and N-Tetradecanoyl-L-Homoserine Lactone were obtained from wound isolates. N-Tetradecanoyl-L-Homoserine Lactone was the only signaling molecule obtained from ear isolates. Further research should be done to develop rapid tests that could be possible to detect acyl homoserine specific to Pseudomonsa aeruginosa present to human samples and deliver results in real time. Signaling molecules inhibitors (SMI) are possible potential therapeutics that could produce the subsequent class of antibacterial drugs. The fundamental role of each signal molecule in the production of biofilms and virulent factors should also be investigated, as well as whether Pseudomonas aeruginosa needs one or more signaling molecules to form quorum sensing. Identification of the signals molecules (AHLs) of P. aeruginosa and developing signaling molecules inhibitors (SMI) can have an important prognostic, diagnostic and therapeutic value in human medicine for the treatment of P. aeruginosa infections.

Preserving Tradition and Promoting Innovation: “Quesillo,” Artisanal Cheese from Oaxaca, Mexico

The quesillo is a traditional food originally produced in the state of Oaxaca, Mexico, is fresh, spun paste or strands, made with raw milk and has unique sensory attributes due to its extensive native microbiota, mainly lactic acid bacteria, which are fermenting microorganisms, safe, are used in the fermentation and preservation of food helping to improve flavor, aroma, and texture. They are part of the natural microbiota of raw milk and its derivatives, mainly traditional cheeses, where the greatest diversity and heterogeneity is found. The objective of this study was to microbiologically identify lactic acid bacteria (LABs) present in the traditional quesillo of Oaxaca, Mexico. Quesillo samples were collected in three municipalities in Oaxaca, Mexico. Serial dilutions were prepared from 10 g of sample, then seeded in a Man, Rogosa, and Sharpe (MRS) selective medium. Subsequently, they were incubated at 30 °C for 24 to 48 h. The strains were identified macroscopically and microscopically, catalase and oxidase tests were performed as confirmation tests for LAB. From the three traditional quesillo samples, 86 colonies with BAL macroscopic morphology were identified, strains with yeast morphology, coccobacillary, catalase and oxidase positive were discarded, and finally 56 strains were identified with macroscopic and microscopic characteristics of LAB type, gram positive, bacillary and coccobacillary morphology, catalase and oxidase negative. In conclusion, quesillo is a traditional product that presents lactic acid bacteria as important microorganisms, which, in addition to improving sensory attributes and food preservation, can present probiotic properties that when administered in adequate amounts confer a health benefit to the host.

Microbiological and Chemical Characteristics of Yoghurt Incorporated with Watermelon Rind Powder

Watermelon rind accounts for approximately one-third of the overall fruit mass. It is usually discarded due to its low commercial value. However, it is reported to contain valuable nutrients and is an effective source of pectin that can act as a potential prebiotic. This study aimed to study the effects of watermelon rind powder (WRP) on the growth of probiotic bacteria in yoghurt and its chemical characteristics. Watermelon rind was dried by using a dehydrator and ground into powder form before being incorporated into fresh yoghurt at 2% and 4% w/v. A sample with 0% w/v WRP was prepared as control. The effect of WRP on the growth of probiotic bacteria was determined by MRS plate count. Chemical analyses including titratable acidity, pH and Brix were conducted during the fermentation process. The results showed that the increase in WRP percentage resulted in a significant increase in bacterial growth with 7.20 ± 0.22 log CFU/mL for the control sample as compared to 8.42 ± 0.23 log CFU/mL for sample with 4% WRP after 30 hr of incubation. The fermentation time was also improved with the presence of WRP with a 0.22 h-1 increase in growth rate observed for the sample with 4% WRP as compared to the control sample. Furthermore, samples containing 4% WRP showed the highest increment in titratable acidity (12.47) and the highest percentage in Brix value reduction (51.04%) during the fermentation period as compared to the control sample. Biochemical analysis showed negative values for oxidase and catalase test while positive values were obtained for gram-staining indicating the presence of Lactic acid bacteria from the gram-positive group. This study demonstrates the high potential of WRP in promoting bacterial growth for yoghurt production which is beneficial to the food industry other than promoting the ongoing effort of food waste reduction.

Staphyloxanthin and Meropenem Effects on Some Capsular Genes Expressions of Klebsiella pneumoniae Isolated from UTIs

Klebsiella pneumoniae has become one of the most common etiologic agents in the context of recurrent UTI. Capsule is the earliest recognized factor determining bacterial pathogenicity. This study aimed to test the effects of staphyloxanthin, meropenem and a combination of them on capsule genes of Klebsiella pneumoniae. Eighty-eight gram negative bacterial isolates were collected from 150 urine specimens between September and November of 2022.after cultivation on MacConkey agar medium. Oxidase (negative) and catalase (positive) tests were performed. In addition, all isolates were found to be non-motile when their motility was further examined. The results presented that 77 (51.3%) could ferment lactose (lactose fermenter), while 30 isolates (20%) of them were confirmed as K. pneumonia. The antibiotics sensitivity test of (30) K. pneumoniae revealed that 28 (93%) of isolates were sensitive to meropenem, and about 20 (67%), 16 (53%), 8 (27%), 7 (23%), 5 (17%) and 4 (13%) were sensitive toward amikacin, ciprofloxacin, trimethoprim, cefepime, tetracycline and Augmentin respectively. S. aureus (producers of staphyloxanthin) isolates were taken; the staphyloxanthin pigment was extracted, purified by Millipore filter (0.22 mm). Conventional PCR results revealed the presence of galF gene (141 bp) in all eight tested isolates (100%). In addition, the wzi gene (177bp) was found in seven isolates (87.5%). According to the quantitative real-time PCR results, most treatments resulted in downregulation of galF and wzi genes expression, whereas some treatments resulted in upregulation.

Genotypes and Phylogenetic Analysis of Helicobacter pylori Clinical Bacterial Isolates

Helicobacter pylori is a human pathogen bacterium associated with gastritis, peptic ulcer, and gastric cancer. It can be identified through the 16S rRNA gene and characterized through cagA and vacA virulence genes. Clinical cultures of H. pylori were isolated and identified from human stomach biopsies. The isolates were characterized according to their colonial and microscopic morphology, and molecular genotyping was conducted to determine the bacterial virulence. A phylogenetic analysis of the 16S rRNA gene sequencing was performed. In addition, multilocus sequence typing analysis was performed to determine the phylogeographic nature of the isolated strains. Three bacterial isolates were selected from 22 gastric biopsies, identified as H. pylori through colonial morphology, Gram staining, urease, catalase, and oxidase tests and identification of the ureC gene through end-point PCR. Amplification of 16S rRNA, urea, and tonB genes was performed, as well. Differences between the cagA and vacA genotypes were determined among the isolates. The phylogenetic analysis confirmed the identity of the three isolates as the specie Helicobacter pylori. Different genotypes were obtained for each H. pylori strain, and all the clinical isolates showed the vacA s2/m2 genotype, indicating an absence of the VacA cytotoxin. Only HCGDL-MR01 is a cagA gene carrier with a greater risk to develop a serious disease, such as stomach cancer and peptic ulcer. The multilocus sequence typing placed all the strains within the hpEurope population structure.

Isolation and Characterisation of Pink Pigmented Facultative Methylotrophs (PPFM) from Malaysian Ulam Leaves

This study aims to isolate and characterise the Pink Pigmented Facultative Methylotrophs (PPFM) bacteria from the leaves of common Malaysian table salad (ulam). The colonies of PPFM bacteria were obtained using selectively modified Pseudomonas agar based on the appearance of the pink pigment colony. The three selected isolates labelled OJ4, OJ154 [Oenanthe javanica (Blume) DC.] and ML8 [from Melicope lunu-ankenda (Gaertn.) T. G. Hartley] were chosen for characterisation. The result showed the PPFM bacteria colonies (log CFU/g) at the leaf surface of C. caudatus are 4.4 ± 0.1, significantly higher than O. javanica, 3.8 ± 0.2, and M. lunu-ankenda, 3.2 ± 0.1. The selected isolates belong to the Gram-negative group, motile with rod shape with size [length (l) × width (w)] in µm unit 4.3 ± 1.1 × 1.6 ± 0.6, 5.4 ± 0.2 × 1.2 ± 0.1, and 3.5 ± 0.7 × 1.0 ± 0.0, respectively. They show positive urease, catalase, and oxidase activities, while none of them can degrade starch, gelatine, or cellulose, as well as glucose fermentation (MR test) and metabolism actions (VP test), producing indole and hydrogen sulphide gaseous. Only isolate OJ154 demonstrates positive casein hydrolysis and nitrate reduction activities, while only isolate ML8 can utilise citrate but not in lipid degradation. Their sequence analysis of the 16S rRNA indicated that OJ4 and ML8 are Methylobacterium radiotolerans with a similarity of 99%, whereas, OJ154 is Methylorubrum salsuginis with a similarity of 99%. To conclude, PPFM bacteria from the leaves of C. caudatus, O. javanica, and M. lunu-ankenda have been isolated and characterised in particular.

Isolation and Characterization of Azotobacter and Phosphate Solubilizing Bacterial Isolates

Isolation from 20 soil samples collected from rhizosphere of watermelon was done on Ashby's medium and Pikovskaya's agar media.8 Azotobacter and 6 Phosphate solubilizing bacteria PSB isolates were obtained from all samples. All the obtained isolates of Azotobacter sp and PSB were gram negative. On morphological study it was observed that, all Azotobacter isolates were motile and positive for KOH test. In case of PSB, the colony shapes of 4 obtained isolates were circular and irregular in 2. All the isolates showed smooth appearance on the media with white colony colour. The biochemical results for both Azotobacter spp and PSB revealed that, most of them were positive for methyl red test, catalase test, starch hydrolysis, gelatine hydrolase test, oxidase test and indol test. Out of eight Azotobacter isolates, Isolate 4 (Azob-1) showed highest (14.25 mg/ml) N fixing ability whereas, from of six PSB isolates, Isolate1 (PSB-1) showed highest solubilising index (4.13) therefore these two isolates were found most efficient in their roles and therefore recommended for use for field experiments.

Neisseria gonorrhoeae isolates resistant to extended spectrum cephalosporins and macrolides isolated from symptomatic men in western Kenya.

We characterized the antimicrobial resistance (AMR) profiles of Neisseria gonorrhoeae (NG) isolated from symptomatic men at a sexually transmitted infection clinic in Kisumu, Kenya. Two urethral swabs were obtained from symptomatic men between 2020 and 2022, one for Gram's stain and the other inoculated directly onto modified Thayer-Martin media containing 1% VCNT and 1% IsoVitaleX enrichment. Culture results were confirmed by colony morphology, Gram's stain and oxidase test. Duplicate isolates were shipped to Uniformed Services University for confirmation and characterization. Susceptibility to eight drugs was assessed by E-test. Agar dilution confirmed resistance to ceftriaxone, cefixime, and azithromycin. Susceptibility, intermediate resistance (IR), and resistance (R) were determined according to published criteria. Of 154 enrolled participants, 112 were culture-positive for NG. Agar dilution results in 110 (98.2%) showed the following: azithromycin-R (1.8%), and 4.5% R or IR to ceftriaxone or cefixime: ceftriaxone-R (0.9%), ceftriaxone-IR (2.7%), and cefixime-IR (2.7%). By E-test, most isolates were IR or R to tetracycline (97.2%), penicillin (90.9%), and ciprofloxacin (95.4%). We detected NG with resistance to azithromycin and ceftriaxone, indicating a growing threat to the current Kenyan dual syndromic treatment of urethritis with cephalosporin plus macrolides. Ongoing AMR surveillance is essential for effective drug choices.

Prevalence and antimicrobial susceptibility pattern of Vibrio cholerae isolates from cholera outbreak sites in Ethiopia

BackgroundCholera is an acute infectious disease caused by ingestion of contaminated food or water with Vibrio cholerae. Cholera remains a global threat to public health and an indicator of inequity and lack of social development. The aim of this study was to assess the prevalence and antimicrobial susceptibility pattern of V. cholerae from cholera outbreak sites in Ethiopia.MethodsAcross-sectional study was conducted from May 2022 to October 2023 across different regions in Ethiopia: Oromia National Regional State, Amhara National Regional State and Addis Ababa City Administration. A total of 415 fecal samples were collected from the three regions. Two milliliter fecal samples were collected from each study participants. The collected samples were cultured on Blood Agar, MacConkey Agar and Thiosulfate Citrate Bile Salt Sucrose Agar. A series of biochemical tests Oxidase test, String test, Motility, Indole, Citrate, Gas production, H2S production, Urease test were used to identify V. cholerae species. Both polyvalent and monovalent antisera were used for agglutination tests to identify and differentiate V. cholerae serogroup and serotypes. In addition, Kirby-Bauer Disk diffusion antibiotic susceptibility test method was done. Data were registered in epi-enfo version 7 and analyzed by Statistical Package for Social Science version 25. Descriptive statistics were used to determine the prevalence of Vibrio cholerae. Logistic regression model was fitted and p-value < 0.05 was considered as statically significant.ResultsThe prevalence of V. cholerae in the fecal samples was 30.1%. Majority of the isolates were from Oromia National Regional State 43.2% (n = 54) followed by Amhara National Regional State 31.2% (n = 39) and Addis Ababa City Administration 25.6% (n = 32). Most of the V. cholerae isolates were O1 serogroups 90.4% (n = 113) and Ogawa serotypes 86.4% (n = 108). Majority of the isolates were susceptible to ciprofloxacin 100% (n = 125), tetracycline 72% (n = 90) and gentamycin 68% (n = 85). More than half of the isolates were resistant to trimethoprim-sulfamethoxazole 62.4% (n = 78) and ampicillin 56.8% (n = 71). In this study, participants unable to read and write were about four times more at risk for V. cholerae infection (AOR: 3.8, 95% CI: 1.07–13.33). In addition, consumption of river water were about three times more at risk for V. cholerae infection (AOR: 2.8, 95% CI: 1.08–7.08).Conclusionour study revealed a high prevalence of V. cholerae from fecal samples. The predominant serogroups and serotypes were O1 and Ogawa, respectively. Fortunately, the isolates showed susceptible to most tested antibiotics. Drinking water from river were the identified associated risk factor for V. cholerae infection. Protecting the community from drinking of river water and provision of safe and treated water could reduce cholera outbreaks in the study areas.

Morphological and biochemical identification of Ralstonia solanacearum strains in ginger (Zingiber officinale Roscoe) wilt disease

The Ralstonia solanacearum Species Complex (RSSC) is a significant plant pathogen affecting various agricultural commodities, including ginger. Understanding the distribution and characteristics of RSSC in ginger plants is crucial for effective disease management. This study aims to determine the distribution of RSSC in ginger plants and identify the pathogen through morphological and biochemical characterization. The research was conducted in Banyubiru and Getasan districts, Semarang Regency, Indonesia. The study involved purposive sampling, isolation, and purification of isolates, followed by morphological characterization through observation and biochemical characterization using Gram staining with KOH, oxidase test, arginine activity test, hypersensitivity test, bacterial density calculation, Koch's postulates, and biovar characterization. Three isolates from Banyubiru displayed typical morphological characteristics of RSSC, including round, mucoid colonies with red centers and white edges on TZC medium, and rod-shaped bacterial cells. Biochemical characterization identified these isolates as RSSC strains biovar 3 and 4, capable of causing bacterial wilt in ginger plants. The study confirmed the presence of bacterial wilt in ginger in Banyubiru district. The findings reveal the spread of bacterial wilt caused by RSSC in ginger plants in Banyubiru, Semarang Regency. There is a need for measures to control the potential spread of RSSC in the surrounding host plant areas in Tlumpak Village, Banyubiru district.

Isolation And Identification of Bacteria Associated With Bioelectricity Generation From Fruit Wastes And Gutter Sludge Using Microbial Fuel Cells (MFC) In Kano Metropolis, Kano State, Nigeria

Bioelectricity generation is another environmentally friendly mean of waste management, commonly carried out using microbial fuel cell (MFC) technology. The present work aims to generate bioelectricity from the fruit wastes using microbial fuel cells (MFC) technology and to isolate and identify bacteria during bioelectricity production. Fruit samples from different fruit wastes of banana, lemon, orange, water melon and pawpaw were collected from Na’ibawa-Yanlemo fruit market’s dump site in Kano metropolis, Kano State Nigeria. Physicochemical parameters such as biological oxygen demand (BOD), chemical oxygen demand (COD), total carbon (TC), total nitrogen (TN), organic matter (OM), Temperature and pH of slurries of individual and mixed fruit wastes were determined. Biochemical tests such as voges-proskauer test, oxidase test, catalase test, citrate test and sugar fermentation (TSI) test was also carried out using standard procedures. Generation of bioelectricity was monitored and recorded within a constructed seven dual chambered microbial fuel cells (MFCs) with wasted fruit as readily available materials. Current, current density, power and power density produced from different waste samples for twenty days was also recorded as bioelectricity generated from microbial fuel cells. Bacterial counts at the beginning and end of bioelectricity generation was also recorded from different fruits waste samples for microbial fuel cells (MFCs). The result of pH mean value of slurries of individual fruits is recorded to be all acidic with lemon having the highest acidity recorded as 3.45±0.62. The pH mean value of the mixed fruit was recorded as7.1±0.23. Waste pawpaw fruit had 6±0.62 % the percentage of organic matter among all the individual waste fruits while the mixed fruit combined had 10±0.62 % mean value of percentage organic matter (OM). Two (2) bacterial genera were biochemically identified as Klebsiella pneumoniae and Esherichia coli. The highest generation of voltage (mV) from microbial fuel cells was achieved from gutter, mixed fruit and orange as 650, 400 and 390 mV respectively. The peak highest current produced was achieved from orange waste samples as 0.7mA/m2 at day fourteen. Waste lemon had the highest colony count among all the fruits at the beginning and end of the bioelectricity generation of 2.2×103 and 4.12×102 respectively. Mixed fruits had the colony count of 2.10×105 and 7.20×103 at the beginning and end respectively.

Antimicrobial resistance pattern of Klebsiella species isolated from various clinical specimens

Objective: The present study was carried out to identify Klebsiella species and their antimicrobial resistance pattern from various clinical specimens. Methods: This cross study was conducted at a tertiary care hospital, Dhaka, Bangladesh from July2020 to June 2021. Written consent was taken from the concerned authority. A total 6739 clinical samples including urine, wound swab/ pus, sputum, blood and endotracheal aspirates were collected and processed for isolation of bacteria by following standard guidelines from adult patients. All the urine, wound swab, blood and endotracheal aspirates were inoculated in both blood agar and MacConkey agar media, for blood samples blood culture was done in BacT/ALERT 3D 60 aerobic bottles irrespective to antibiotics administration. After incubation at 370C aerobically for 24 hours incubated plates were examined. Phenotypic identification of the organisms were done by observing colony morphology, hemolytic properties, pigment production, Gram staining and biochemical tests (oxidase test, catalase test and biochemical reactions after inoculation in TSI, MIU, citrate agar media). The Kirby - Bauer disk diffusion method was used for antimicrobial susceptibility testing of the isolated organism using Mueller-Hinton agar and commercially available antibiotic discs (Oxoid Ltd, UK). The data were analyzed using SPSS software Version-20 (SPSS Inc., Chicago, IL, USA). Categorical variables were expressed as numbers (n) and percentages (%). Results: A total 6739 samples were included in the present study. Of which, 170 were urine, 162 were wound swabs, 71 were sputum, 52 were endotracheal aspirates and 45 were blood samples. Culture positivity among the samples is shown in table I. Conclusion: Most of the Klebsiella species are highly resistant to commonly used antibiotics. This higher resistance is the major cause morbidity and mortality related with infections in hospitalized patients. So, a regular surveillance of antibiotic susceptibility pattern is needed to prevent indiscriminate use of antibiotics. CME J 2024; 3(2); 12-15

Genetic Basis of Biofilm Formation Genes Ebp and Bph (phos) among Multidrug Resistance Enterococcus faecalis Isolates, Iraq

Abstract Background: Biofilm produced by Enterococcus spp. increase their inherent and acquired antibiotic resistance, posing a significant challenge to infection treatment, particularly in virulent strains. Objective: This study aimed to investigate some genes responsible for biofilm formation (Bph (phos) and EbpB) by polymerase chain reaction (PCR) technique. Materials and Methods: A total of 130 clinical samples were collected during this study, which were obtained from patients who were admitted to Feminine and Children Teaching Hospital in Al-Diwaniyah City during a period extending from (November 2022 to March 2023). All samples were subjected to culturing on different media (blood agar, MacConkey, Bile Esculin agar, and M-El chromogenic ager), after which catalase and oxidase tests were conducted biochemically. After cultivation, the VITEK-2 compact system was used to identify the samples. The isolates were investigated genotypically for harboring biofilm formation genes, including EbpB and Bph (phos), by molecular method (PCR). Results: The VITEK-2 compact system revealed that only 12 isolates were identified as E. faecalis. The most effective antibiotics against E. faecalis were (vancomycin, teicoplanin, linezolide, tigecycline, and ampicillin), and the highest resistance was against erythromycin (100%) and tetracycline (91.6%). Enterococcus faecalis isolates were investigated genotypically for harboring biofilm formation genes that include EbpB and Bph (phos) by molecular methods, PCR. The results showed that EbpB and Bph (phos) genes were positive for all isolates with a percentage of 100%. Conclusions: It was found that the presence of biofilm formation genes in E. faecalis EbpB, and phos (Bph) increase the pathogenicity of this pathogen. These genes showed a high percentage among E. faecalis isolates. Also, all isolates had the potential to form a biofilm, which complicates their treatment with antibiotics, confirming the critical need to develop novel antimicrobial agents that control the infection associated with the development.

Roseibium algae sp. nov., isolated from a marine alga (Grateloupia sp.).

A novel Gram-stain-negative, rod-shaped, non-spore-forming, aerobic, motile bacterium with a single polar or subpolar flagellum, designated strain H3510T, was isolated from marine alga collected on sea shore of Yantai, PR China. The organism grew optimally at 28 °C and pH 7.0 and in presence of 3.0 % (w/v) NaCl. The strain exhibited positive catalase activity but negative oxidase and nitrate reduction activities. The predominant cellular fatty acids were C18 : 1 ω7c and/or C18 : 1 ω6c, 11-methyl C18 : 1 ω7c, and C16 : 0. Additionally, the major polar lipids were phosphatidylglycerol, phosphatidylmonomethylethanolamine, diphosphatidylglycerol, and phosphatidylethanolamine; the respiratory quinone was ubiquinone 10 (Q-10). The genomic DNA G+C content of strain H3510T was 54.2%. The novel strain showed the closest relationship with Roseibium polysiphoniae KMM 9699T with 98.2 % 16S rRNA gene sequence similarity. The calculated values for average nucleotide identity and DNA-DNA hybridization between strain H3510T and the phylogenetically related Roseibium species were in the range of 71.3-74.9 % and 13.7-19.9 %, respectively. Based on polyphasic analyses, strain H3510T was identified as representing a novel species of the genus Roseibium, for which the name Roseibium algae sp. nov. is proposed. The type strain is H3510T (=KCTC 8206T=MCCC 1K04325T). The heterologously expressed inositol 2-dehydrogenase gene from strain H3510T displayed high oxidation activity on myo-inositol and showed potential in the production of rare stereoisomers of inositol, such as scyllo-inositol.

Morphogenetic characterization of Stenotrophomonas maltophilia infecting white stripe disease of rice (Oryza sativa L.)

Rice is a major cereal crop which ensure food security to more than half of the global population. Several biotic factors impact rice grain quality and its final production. White stripe disease, caused by pathogen Stenotrophomonas maltophilia is considered among the major limiting factor for reducing rice yields and quality. Present study was performed to understand the white stripe disease, which has been frequently misdiagnosed as bacterial leaf blight (BLB) due to similar symptoms. A survey was carried out based on accessibility and farmer participation to monitor incidence and sample collection. The survey was conducted in districts Faisalabad, Gujranwala, Sialkot, Sheikhupura, and Hafizabad, these districts were selected for their importance for rice cultivation in Pakistan. The total sample size was around 500 leaves distributed evenly throughout each study area. The results of study indicated presence of new pathogen of rice. These isolates were biochemically identified and confirmed by gram staining negative, 3% KOH positive, 5% salt tolerance positive, oxidase test negative, catalase test positive, starch hydrolysis test negative, nitrate reductase test positive, indole test negative, lactose test positive, maltose test positive, methyl red test negative, Voges-prokauer test negative and urea hydrolysis test negative. The pathogenicity test was confirmed that pathogen and the Sialkot isolate were the most aggressive isolate among the five isolates collected form the studied areas. The molecular characterization was accomplished by PCR and sequencing. The results of the phylogenetic study indicate that this pathogen belongs to a distinct group, as it is distantly related to Xanthomonas oryzae pv. oryzae (Xoo). This study provides important findings into a novel clade of pathogen causing white stripe disease in rice.

In Vitro Antibacterial Activity of Aloe Barbadensis Millar (Aloe Vera) Against Pseudomonas aeruginosa Isolates in Khartoum State, Sudan

Background: It is becoming more widely acknowledged that Pseudomonas aeruginosa (P. aeruginosa) is a significant nosocomial pathogen that can cause serious infections. In general, new therapeutic approaches that prioritize the utilization of extracts and physiologically active chemicals extracted from herbal plants should be used to control P. aeruginosa infections. The World Health Organization (WHO) estimates that in underdeveloped countries, around 80% of the population mostly uses traditional medicine and plant extracts as their primary source of medicine to treat a variety of infectious diseases. Objective: The purpose of this study was to investigate the in vitro antimicrobial activity of A. vera against P. aeruginosa isolated in Khartoum state hospitals. Materials and Methods: The present descriptive study was performed in Khartoum state. Between November 2022 and April 2023, a total of 100 clinical isolates of P. aeruginosa were obtained from patients hospitalized at Khartoum State hospitals. Standard microbiological techniques, such as the Gram stain, catalase test, and oxidase test, were used to identify the isolates. Ciprofloxacin, Ceftriaxone, Meropenem, and Gentamicin. A. vera gel were used in an antimicrobial susceptibility test against the P. aeruginosa strains. Results: Isolates of P. aeruginosa were collected from hospitalized individuals. Of the 100 isolates included in the study, 63 isolates (63%) had been isolated from males and 37 isolates (37%) had been isolated from females. Using the microdilution method, antimicrobial susceptibility testing for 100 P. aeruginosa clinical isolates was conducted for four antibiotics. The results showed that 38% of the isolates were resistant to ciprofloxacin, 94% to ceftriaxone, 82% to gentamicin, and 0% to meropenem. Conclusions: P. aeruginosa demonstrated multidrug resistance to a widely used antibiotic. A different therapeutic agent for the treatment of clinical multi-drug resistance bacteria may be employed in the ethanol extracts of Aloe vera since they have shown antibacterial efficacy against multi-drug resistant P. aeruginosa which is resistant to many drugs and may be treated with a combination of conventional antibacterial discs and the crude extract of Aloe vera. The study's findings validate the traditional wisdom that aloe vera plants may cure microbial infections and suggest that aloe vera may be used to make powerful new antibacterial compounds.