Oxidase-positive Research Articles

Enterococcus faecium from chicken feces improves chicken immune response and alleviates Salmonella infections: a pilot study.

Probiotics reduce the emergence of antibiotic resistance in the livestock industry. Chicken feces are reservoirs of beneficial microbiomes. The aim of this study was to isolate putative probiotics from the intestinal contents of broiler chickens. Five fecal samples were collected from two poultry farms in Al-Ahsa, Saudi Arabia. Of the 11 morphologically distinct isolates from chicken feces (CF) samples, five isolates displayed positive reactions to Gram staining, catalase, and oxidase tests, and reacted negatively to a hemolytic assay. The isolates CF1, CF2, CF3, CF8, and CF11 were selected for further analysis of probiotic characterization, gastric survival capacity, antibiotic susceptibility, and antimicrobial activity against poultry infected with Salmonella enterica. CF2 and CF11 showed the highest hydrophobic values (> 51% hydrophobic nature). CF1, CF2, and CF11 showed potent antimicrobial activities. The active isolate CF2 was identified as Enterococcus faecium by 16s rRNA sequencing and showed a genetic similarity of 99.1%. An in vivo study was conducted using a chicken model. Enterococcus faecium-fed chickens showed an improved body weight and a lower mortality rate (17-34%). Salmonella enterica colony-forming unit (CFU) invasion in the spleen and thymus was significantly reduced in the E. faecium-fed chickens. The fecal S. enterica load was reduced from CFU 6.8 to 3.9/g in oral-administered E. faecium-fed chickens. Enterococcus faecium-fed chickens showed increased levels (P < 0.01) of butyric acid and reduced levels (P < 0.01) of intestinal interleukin 1 beta, C-reactive protein, and interferon gamma levels compared to those in the S. enterica-infected chicken group. In addition, E. faecium showed significant binding to Caco-2 epithelial cells in vitro and inhibited S. enterica colonization, indicating co-aggregation of E. faecium in epithelial cells. These results revealed that an E. faecium formulation could prevent bacterial infection and improve the quality of broiler chickens.

A STUDY ON CLINICO-MICROBIOLOGICAL PROFILE OF CONJUNCTIVAL FLORA IN CONGENITAL DACROCYSTITIS

Dacryocystitis is an inammation of the lacrimal sac and duct. It may be congenital or acquired.It is an important cause of ocular morbidity both in children and adults. To isolate &amp; identify the causati PURPOSE- ve bacteria from clinically diagnosed cases of congenital dacryocystitis &amp; to study the organism in relation to clinical prole. -Gram฀s stain MATERIALS &amp; METHODS &amp; Giemsa stain ,Culture media,10 % KOH preparation , MacConkey฀s agar, 5% Sheep Blood agar, Chocolate agar .Biochemical tests on Isolated colonies from the plates were identied by Colony morphology, Gram฀s stain, Oxidase test, Catalase test, and Motility.For further identication isolated colony was inoculated into peptone water and Hiss serum for fastidious bacteria, which was incubated aerobically and inoculated into appropriate medias for biochemical tests like Indole production, Methyl red, Voges-proskauer test, Citrate utilization, Urease hydrolysis, Sugar fermentation, Triple Sugar Iron agar, Nitrate reduction test and Amino acids decarboxylase tests. The specic tests like Inulin fermentation, Optochin sensitivity test and Bile solubility test were done. Antimicrobial susceptibility tests: The standardized Kirby-Bauer disc diffusion test of the Clinical and Laboratory Standards institute (formerly NCCLS) was used for testing. The media used was Mueller-Hinton agar for non-fastidious organisms. 5% Sheep Blood agar added to Mueller-Hinton agar for fastidious organisms and Haemophilus Test Medium was used for testing Haemophilus inuenzae and it was read after 16-18 hours. The zone of inhibition was measured and reported as Susceptible or Resistant. For detection of methicillin resistant Staphylococcus aureus detection disc tests for methicillin resistance was done using 10μg oxacillin disc on Mueller-Hinton agar containing 4% sodium chloride. In our study , femal RESULTS- es are more (7) as compared to males (4)&amp; the culture positive cases were 10 and in 1 case culture showed no bacterial growth. The most frequent Gram-positive isolates are Streptococcus pneumoniae 7 (63.63%) followed by Staphylococcus aureus 1 (9.09%) and the most frequent Gram-negative isolates are Pseudomonas aeruginosa 3 (27.27%). In our study the Gram-positive isolates were most sensitive to CONCLUSION- Vancomycin (100%), followed by Tobramycin and Linezolid (97.14 %). Least sensitive antibiotic against Gram-positive organisms was Penicillin (51.42 %). The Gram-negative organisms were most sensitive to Tobramycin and Gentamicin (100%), followed by Amikacin, Oaxacin (92.59 %), Cefepime (88.88 %). Least sensitive antibiotic against Gram-negative organisms was Ciprooxacin (77.77 %)

EXPLOITING RHIZOBIUM, AZOTOBACTER, AZOSPIRILLUM AND ACETOBACTER BACTERIA AS BIO-FUNGICIDE AGAINST WILT CAUSING FUNGI

The current study was carried out at the Department of Agricultural Microbiology, College of Agriculture, Raipur, C.G., during the years 2021–2022. This study was set up with the general objective of investigating the interaction between Fusarium and plant nutrient mobilizing bacterial isolates namely Rhizo-L3, Rhizo-L4, Azoto-MW15, Azoto-Palak16, Azos-Palak13, Azos-Palak17, Aceto-Root18 and AcetoRD15. These isolates were selected and tested to control wilt disease causing fungi. In this connection, these isolates were tested for different biochemical parameters viz.starch hydrolysis, catalase, urease, oxidase test etc. Further these isolates were evaluated for their ability to inhibit the mycelial biomass of wilt causing fungi Fusarium oxysporium and Sclerotium rolfsii in modified Martin broth and agar media. Results show that the maximum growth inhibition of Fusarium oxysporium (83.62%) Azoto-MW15 + Azoto-Palak16 exhibited followed by mixed cultures of Rhizo-L3 and Rhizo- L4 (79.40%), Aceto-RD15 + Aceto-Root18 (74.39 %) and Azos-Palak13 + Azos-Palak17 (72.99 %), while, in plate assay with Sclerotium rolfsii also showed inhibition of 56.44%, 53.19% and 52.21 % respectively.

INFLUENCE OF FOLIAR SPRAY OF BIO-INOCULANT AND MICRONUTRIENT ON SOIL BIOLOGY AND YIELD IN BROCCOLI

The experiment was carried out in the years 2021-22, at the Department of Agricultural Microbiology, Indira Gandhi Krishi Vishwavidyalaya, Raipur (Chhattisgarh). The study involved application of foliar spray of bio-inoculants and micronutrient; Ammonium Molybdate. Bio-inoculants; Azotobacter and Azospirillum inoculation strains were collected from Microbiology repository and were revived for biochemical characteristics on Jensen’s and Okon’s media respectively. Both the isolates showed positive reaction for starch hydrolysis test, catalase test, urease test and oxidase test. Plant parameters evaluated include biomass accumulation and fruit weight per plant. The experimental results revealed that significantly maximum fruit weight per plant (359.0g), dry shoot biomass (16.6g/ plant), dry root biomass (8.6g/ plant) were recorded under T4 (IF + Foliar Application of Bio-inoculants and Ammonium molybdate 0.2%). Besides higher microbial population, Azotobacter (44.3 x 103 cfu/ g of soil), Azospirillum (28.5x 103 cfu/ g of soil) were also observed to be significant under T4. Dehydrogenase activity in rhizosphere soils was found maximum 27.3 µg TPF/g soil/h under T4 (IF + Foliar Application of Bio-inoculants and Ammonium molybdate 0.2%) at 60DAT. Thus application of Bio-inoculants along with Ammonium molybdate (0.2%) as foliar spray along with 50% inorganic fertilizer proved to be the most effective and may be adopted in Broccoli cultivation.

Emergence of Multidrug Resistant Vibrio cholerae O139 in Acute Diarrhoea Patients Attending a Tertiary Care Hospital, West Bengal, India

Introduction: India is a developing country with many poor sanitation areas. Cholera, a water borne disease is rampant in areas of poor sanitation and is mainly due to Vibrio cholera of O1 and O139 serogroups causing acute cases of rice watery diarrhoea and high mortality. Infection due to Multidrug Resistant (MDR) Vibrio is on the rise. It is endemic in over 50 countries including India leading to a number of epidemics and pandemics. Till date, about seven pandemics have been identified due to cholera infection. Aim: To isolate Vibrio cholerae from acute diarrhoea cases with their antibiotic susceptibility and identify MDR strains, if any. Materials and Methods: A hospital-based, cross-sectional study was conducted in the Department of Microbiology, Burdwan Medical College, Purba Burdwan, West Bengal, India, from January 2021 to December 2021. Rectal swabs/faeces collected in Cary-Blair Transport Media were brought to the laboratory and incubated at 37°C for six hours; a loopful was inoculated in nutrient agar, MacConkey agar and selective media Thiosulphate-Citrate-Bile Salts-Sucrose agar (TCBS). Gram stain, motility, colony characteristics, oxidase test, cholera-red reaction and slide agglutination test was done. Diagnosis was confirmed; antibiotic susceptibility done and interpreted as per Clinical and Laboratory Standard Institute (CLSI) guidelines. The data was collected and entered into Microsoft Excel software and presented as frequency and percentages. Results: Total 60 samples were collected and tested, out of which 24 (40%) were positive for Vibrio cholerae and nine in 24 (37.5%) samples were MDR strains. Twelve samples were from femlaes and 12 were from males, majority {5 (41.67%) females, 9 (75%) males} belonged to the age group of 0-5 years. Serotyping revealed that Vibrio cholerae O1 serogroup was identified in 8 (33.33%) cases and O-139 serotype in 16 (66.67%) samples. Rest were Escherichia coli (n=10, 25%), Klebsiella pneumoniae (n=5, 12.5%) and Staphylococcus aureus (n=1, 25%). Conclusion: There is a rise of infections due to MDR strains of Vibrio O139 sero group in the community which needs early diagnosis and treatment for control. Vibrio cholerae strains were more resistant to fluoroquinolones, macrolides, tetracycline and Ampicillin.

Phenotypic Characterization of Carbapenemase-Producing Enterobacteriaceae Strains in a Referral Teaching Hospital in Yaound&amp;#233;, Cameroon

Background: Carbapenemase-producing Enterobacteriaceae (CPE) are an important and increasing threat to global health. They are nowadays more encountered routinely in hospitals and cause high morbidity and mortality due to limited therapeutic alternatives. This study sought to determine the prevalence of CPE in Yaoundé teaching hospital, Cameroon, and the associated risk factors. Materials and Method: To achieve this goal, a descriptive cross-sectional study coupled to an analytical component with consecutive collection of Enterobacteria strains was carried out during a three-month period (from 27th July to 24th October 2018) in the University Teaching Hospital of Yaoundé, Cameroon. The oxidase and biochemical identification tests using a miniaturized Api 20 E system were performed on colonies grown on Eosin Methylene Blue (EMB) medium and subcultured on nutrient agar. Drug susceptibility testing was carried out according to the Antibiogram Committee of the French Society of Microbiology (CA-SFM 2018.V.2.0). The detection of carbapenemase production was performed by the CA-SFM 2018 algorithm for the screening of carbapenemase-producing enterobacteriaceae and its classification by inhibitory synergy tests. Results: Out of the 104 isolates, Escherichia coli (50%) was the most prevalent species, followed by Klebsiella pneumoniae (37.5%) and Citrobacter frendii (12.5%). Drugs susceptibility patterns showed a high resistance to penicillins group (97.4% to amoxicillin), cephalosporins (68.4% to cefotaxim, 58.1% to cefixim, 60.7% to ceftazidim, 57.1% of cefoxitin) and aztreonam (55.7%). However, 11.9% carbapenems related resistance was noticed: 14.4% to imipenem, 13.8% to ertapenem and 7.5% to meropenem. Numerous co-resistance to quinolones (65.8%), fluoroquinolones (49.6%), aminoglycosides (49.6%) and cotrimoxazole (71.8%) were also observed. From 104 isolates, AmpC production represented 23.08% (25/104) and 36.54% (38/104) were ESBL-isolates. The overall prevalence of CPE was 25% (26/104) with K.pneumoniae predominant (61.53%). Besides, Class A and class B carbapenemase were mainly produced with respectively 20% (21/104) and 5% (5/104). Univariate analysis revealed a significant association of carbapenemase production to Klebsiella pneumoniae (p = 0.01), ESBL and AmpC production ((P = 0.01 and P = 0.001 respectively) while that association was only significant to Klebsiella spp (p = 0.04) and AmpC production (p = 0.02) in multivariate analysis. Conclusion: The multi-resistance of Enterobacteriaceae to antibiotics in Cameroon has considerably increased. More attention should be paid to those bacteria to stall antimicrobial resistance spread.

Identification and Prevention of Microbial Contaminants in Musa paradisiaca Tissue Culture

Banana (Musa spp.) is an economically vital fruit crop in Malaysia and therefore, micropropagation was applied to meet the market demand for disease-free seedlings. However, microbial contamination is a significant obstacle in micropropagation techniques. In this research, the microorganisms that were present in the banana culture were characterized and the efficacy of antimicrobial and antifungal agents to inhibit contaminants was evaluated. Three bacterial and two fungal isolates were isolated from the contaminated culture. Klebsiella pneumoniae, Klebsiella quasipneumoniae, and Klebsiella variicola were identified by molecular identification based on the 16S rDNA sequence. The gram-staining method confirmed all three bacteria were gram-negative. Oxidase and catalase tests showed the presence of cytochrome oxidase system and catalase enzyme in all bacteria. The bacteria can also hydrolyze starch, ferment sugars, and reduce sulfur from the amylase test. Phenotypic identification of fungi revealed the presence of conidia and hyphae, indicating the presence of Colletotrichum spp. and Aspergillus spp. In fungi characterization, Colletotrichum gloeosporioides and Aspergillus flavus were detected. Chloramphenicol was identified as an effective antibacterial agent from the disc diffusion method. Fluconazole was a potent antifungal agent by screening the sterilizing agents. The findings may potentially lead the way for the implementation of reducing the contamination rate in banana micropropagation.

The in-vitro Antibiotic Sensitivity Test of Pasteurella multocida Isolated from Layer and Breeder Chickens

The current study aimed to characterize Pasteurella multocida (P. multocida) isolates from layers and breeder chickens in Egypt regarding in-vitro antibiotic sensitivity and resistance pattern. In doing so, spleen, liver, lungs, and heart, were taken aseptically from chickens suffering from a drop in egg production, septicemia, respiratory manifestations, and mortalities between 2016 and 2017. To isolate bacteria, samples were grown on a modified Das medium. Moreover, microscopic appearance and biochemical characteristics were used to identify pure colonies of P. multocida isolates. In the next step, in-vitro antibiotic sensitivity testing was performed on the isolated P. multocida. The findings indicated that P. multocida was found in 36 isolates out of 330 investigated chicken flocks. Small glistering, mucoid, grayish, and dew drop P. multocida colonies were discovered during the culture analysis. Pasteurella multocida isolates were Gram-negative coccobacilli using the microscope. Catalase, indole generation, H2S production, nitrate reduction, and oxidase tests were all positive for the sample; however, methyl red, urease activity, Voge's proskaur, and gelatin liquefaction tests were all negative. They also fermented glucose, mannose, fructose, sucrose, mannitol, xylose, and sorbitol without producing gas but not lactose, arabinose, maltose, inositol, salicin, raffinose, or dulcitol. Isolated P. multocida strains were sensitive to tetracycline, erythromycin, trimethoprim/sulphamethoxazole, norfloxacin, ofloxacin, penicillin, chloramphenicol, and azithromycin, while resistant to ampicillin and clindamycin. Cefoperazone, gentamycin, and streptomycin all showed intermediate sensitivity.

Identifikasi Bakteri Mirip Coliform pada Media Cromocoult Coliform Agar (CCA)

Coliform-like bacteria can be found when examining coliforms in fresh water using the CCA media on membrane filter method. Therefore, it is necessary to characterize and identify coliform-like colonies that grow on CCA medium so that is easier to perform colony count analysis. One hundred ml of fresh water samples were filtered using filter membrane paper and then planted in CCA medium. Two different colonies growing on the media (red (A) and pink-purple (B)) were subjected to oxidase test, gram staining and re-cultured in various growth media such as blood agar, nutrient agar, MacConkey, and B. coli agar. Both colony types were also identified with Vitek MS to determine the species. The results of the examination showed that the red colonies (code A) were coliform-like bacteria, while the pink-purple colonies (code B) were coliform bacteria. The characteristics of coliform-like bacteria are that they produce hemolysis on blood agar and do not ferment lactose. Identification with Vitek MS showed that the isolated coliform-like bacteria was Aeromonas hydrophilla. Therefore, it can be concluded that Aeromonas sp. are coliform-like bacteria that grow on CCA, so further verification is needed when counting membrane filter colonies using CCA media.

Antibiogram Analysis of Salmonella paratyphi A Isolated from Gall Bladder Patients in District Peshawar, Pakistan

Salmonella paratyphi A harbors gall bladder in the human body. It serves as a site of persistence for Salmonella paratyphi A. It is an enteric pathogen which has become resistant to many drugs. Therefore, the current study was designed for the identification and antibiogram analysis of S. paratyphi A, isolated from the gall bladder patients undergone cholecystectomy. It included 250 samples of bile, stone, and tissue of patients. The sampleswere cultured on blood, macConkey, and Salmonella Shigella media. Further identification was carried out by morphological oxidase test and Analytical Profile Index (API) strips, followed by antibiogram analysis of the isolates. In the current study, twenty-eight (11.2%) paratyphi A were isolated including 10 (10%) from male patients and 18 (12%) from female patients. Furthermore, 96 samples were found to be positive formiscellaneous growth including 53 with S. typhi (21.2%), 13 with Escherichia coli (5.2%), 09 with Klebsiella (3.6%), 07 with Providencia (2.8%), 05 with Pseudomonas (2%), 03 with Proteus (1.2%), and 06 with Staphylococcus aureus (2.4%). The distribution and susceptibility pattern of S. paratyphi A isolates was checked in different types of clinical specimens including bile, stones, tissue, bile/stones, bile/tissue, stones/tissue, and bile/stone/tissue. S. paratyphi A was distributed as follows: bile (11), stones (5), tissue (3), bile/ stones (4), stones/ tissue (1), bile/tissue (1), and bile/stones/tissue (3). The results of the antibiogram analysisfound that the isolates of Paratyphi A were resistant to sulfamethoxazole 23 (82.14%), cefixime 23 (82.14%),ceftriaxone (rocephin) 20 (71.42%), augmentin 19 (67.85%), and azithromycin 18 (64.28%). The increased susceptibility of these isolates was towards meronem 28 (100%), imipenem 28 (100%), cefoperazon + sulbactam (sulzone) 25 (89.28%), and amikacin 23 (82.14%). The current study signifies the use of the Antibiogram Analysis of Salmonella paratyphi A most susceptible and effective antibiotic options for gall bladder diseases complicated by S. paratyphi A, which showed resistance to ceftriaxone (rocephin), cefixime, sulfamethoxazole, azithromycin, and augmentin, while sensitivity to meropenem, imipenem, cefoperazone + sulbactam (sulzone), and amikacin. It makes the latter a better choice for treatment against the gall stone disease complicated with S. paratyphi A infection.

Efficacy of zinc oxide nanoparticles and Bifidobacterium bifidum Extraction on anaerobic bacteria isolated from patients with diarrhea

Infectious acute diarrhea may be prevented with probiotics; because they make up the majority of the colonic flora in breastfed newborns and are likely to contribute to the lower incidence of diarrhea in this population, Bifidobacteria are particularly appealing as probiotics agents. The present study was designed to identify anaerobic bacteria, especially C. difficile the main reason for dysentery associated with antibiotics. Detect the ability of each ZnoNPs and B. bifidum to inhibit bacterial growth. During the period from March to October 2019, (100) children and adults who came to Salah al-deen hospital in Tikrit city participated in the study under the supervision of a physician. All samples were transported using a carry Blair if late one or two hours after collection and culturing. The collected models were also cultured on Xylose Lysine Deoxycholate agar, Salmonella Shigella agar, Eosin methylene blue agar, and MacConkey agar. For initiated diagnoses of the Enterobacteriaceae, Blood agar is used to detect beta-hemolytic isolates, recover enteric bacteria other than Enterobacteriaceae, and evaluate the results of oxidase tests. To diagnose bacterium kinds, biochemical reactions and motility tests were used. Impact of ZnoNPs, and B. bifidum antibiotic In vitro. The results of 100 dysentery feces samples were obtained into (60%) samples for males and (40%) for females. Eighty-two positively impacted anaerobically on growth media like Clostridium complicate agar and MacConkey agar (18%) other than bacteria. In contrast, negative samples revealed 10 (55.56 percent) samples for males and 8 (44.44 percent) samples for females. The same stool samples were taken and cultured on Clostridium difficile agar and MacConkey agar under anaerobic and ideal incubation conditions. 15% and 67% of isolates appeared on MacConkey agar of the total number of samples, while 18% showed negative growth. Finally, Zn NPs showed their ability to inhibit Clostridium complicated segregate lean on the condensation 5 mg/ml, and it caused the inhibitory effect on Clostridium to complicate by 10-22 of the diameter of inhibition. The Inhibition Zone Dimeter ranged from 8 to 25 mm for isolates when condensation was utilized at 2.5 mg/ml. According to the findings, the widths of the inhibitory zones for isolates of C. difficile containing B. bifidum supernatant mg/ml ranged from 9 to 24 mm. Keywords: Zinc oxide nanoparticles, Probiotic, Bifidobacterium bifidum, Clostridium difficile

&lt;strong&gt; Correlation between presence of bacillus species and OHI-s in recurrent aphthous stomatitis patients &lt;/strong&gt;

ABSTRACTIntroduction: The approximate prevalence of Recurrent Aphthous Stomatitis (RAS) was 45.42% at Gusti Hasan Aman Dental Hospital Banjarmasin. Bacillus sp. is a bacterium that can form spores that can be transmitted through air, dust, water and food. Poor oral conditions will increase the possibility of bacterial contamination, especially for ulcers found in RAS. This study aimed to analyzed correlation between the presence of Bacillus species and OHI-S in the oral ulceration of RAS patients. Methods: This was an analytical cross-sectional study conducted among Oral Medicine Department outpatients at Gusti Hasan Aman Dental Hospital, Banjarmasin, Indonesia. A total of 38 subjects were examined to assess Oral Hygiene Index Simplified and obtained bacteria samples using the swabs technique. Bacteria were then inoculated in blood agar base media and concurrently presented with gram staining. Bacteria were then identified using the catalase test, urease test, and oxidase test. Results: Bacillus species were predominantly identified in oral ulceration of RAS patients at Gusti Hasan Aman Dental Hospital Banjarmasin. Bacillus sp. (60,5%), Streptococcus sp. (34,2%), Neisseria sp. (2,6%), and Lactobacillus sp. (2,6%) were found in ulcerated RAS lesions. Spearman test resulted in p = 0.862 (p &gt; 0.05), where no significant difference was obtained between presence of bacillus species and OHI-s in recurrent aphthous stomatitis patients. Conclusion: Bacillus species were predominantly identified in oral ulceration of RAS patients a. However, the presence of Bacillus sp. was not associated with OHI-S in RAS patients.Keywords: bacillus sp.; oral hygiene index simplified; recurrent aphthous stomatitis

PROFILING OF ANTIBIOTIC RESISTANT BACTERIA ISOLATED FROM POULTRY LITTER OF COMMERCIAL FARMS IN KHULNA DISTRICT, BANGLADESH

Microbial resistance to antibiotics has become a global threat that interferes with the interaction among humans, the environment, and microbes. Our study sought to detect antibiotic-resistant bacteria in light of the growing interest in using antibiotics in the poultry sector. We collected 24 poultry liter samples from 04 different upazilas of Khulna district (Batiaghata, Dumuria, Paikgachha and Koyra). Initially, 09 bacterial isolates were selected and among them, 77.78% bacteria were found gram positive. Subsequently, to characterize these bacteria, a toal of 10 biochemical tests (methyl red test, MacConkey agar test, indole test, catalase test, triple sugar iron test, mannitol salt agar test, oxidase test, Voges-Proskauer test, citrate test and nitrate test) were carried out in this experiment. Moreover, our study also isolated and amplified bacterial DNA for Sanger sequencing and finally disclosed 09 different antibiotic resistant bacterial species, namely Priestia aryabhattai, Bacillus cereus, Priestia megaterium, Lysinibacillus macrolides, Rossellomorea aquimaris, Mammaliicoccus sciuri, Bacillus wiedmannii, Escherichia coli and Citrobacter freundii. Further, disk diffusion assays were performed following CLSI guidelines and it unveiled the susceptibility and resistant properties of isolates against commonly used 08 antibiotics (penicillin 10U, tetracycline 20µg, nitrofuran 300µg, clindamycin 2µg, azithromycin 15µg, quinolones 5µg, tetracycline 30µg and penicillin 30µg) in poultry farms. All of the bacterial isolates were found resistant to at least one of the antibiotics except C. freundii (isolate 9). Most of the isolates (66.67%) were resistant to nitrofuran, whereas all of them were susceptible to penicillin. Lastly, this study also made an effort to understand the evolutionary relationships of the identified species through a phylogenetic tree. Thus, the findings of this study will help farmers and common people to better understand the risk of developing antibiotic-resistant bacteria as a result of excessive antibiotic usage.

Multi-Drug Resistant Campylobacter jejuni of Poultry Origin in Ado-Ekiti, Nigeria

The occurrence of Campylobacter jejeni in the large intestine of domestic fowls in Ado-Ekiti was assessed using standard microbiological procedures. One hundred faecal swabs were inoculated into modified cefoperazone charcoal deoxycholate agar (mCCDA). Twenty-seven isolates of Campylobacter jejeni were recovered from the birds. Biochemical identification of the isolates was carried out using oxidase and catalase tests. Antibiotic susceptibility test was carried out using standard disc diffusion method as specified by the Clinical Laboratory Standard Institute (CLSI), to the following antibiotics; amoxicillin, cefoperazone, ceftazidime, aztreonam, ceftriaxone, pefloxacin, ciprofloxacin, levofloxacin, enrofloxacin and norfloxacin. The pattern of resistance was as follows; Amoxicillin (66.7%), cefoperazone (48.1%), ceftazidime (66.7%), aztreonam (40.7%), ceftriaxone (74.15), pefloxacin (51.9%), ciprofloxacin (33.3%), levofloxacin (40.7%), enrofloxacin (22.25) and norfloxacin (59.3%). Twenty-three different multiple resistance pattern were observed among the isolates. The high level resistance observed in this study poses significant health risk to the general public, a synergistic collaboration is therefore suggested between public health policy-makers and researchers to curb this ugly trend.

Confirmation of Burkholderia gladioli as the Causal Agent of Bacterial Scab on Gladiolus (Gladiolus spp.) in Ohio.

Ohio is one of the top five floriculture producers in the United States, grossing over $200 million annually (NASS 2019). Within the international floriculture trade, gladiolus cut flowers represent the fifth highest grossing crop (Ahmed et al. 2002). In September 2021, the Ornamental Crops Pathology Lab at the Ohio State University received a gladiolus (Gladiolus spp.) sample of an unknown cultivar from a home garden in Franklin Co., OH where several plants had failed to grow from planted corms or were stunted and displaying symptoms of disease. Bleached, water-soaked spots with necrotic margins along the flowering stems, stunted flowers with partial necrosis, and necrotic bracts were observed on the submitted sample. Bacterial isolations were performed by surface disinfesting small sections of bract tissue from the border of a lesion by soaking in 10% bleach for 30 sec and rinsing twice in sterile water, macerating the tissue in sterile water, and streaking the suspension on nutrient agar (NA) plates. Plates were incubated at 28°C for 48 hours and the resulting colonies were purified by re-streaking a single colony on NA twice. Bacterial colony morphology on NA presented as cream-colored and shiny with an irregular form and undulate margin. Five in vitro tests were performed using one representative isolate to identify the bacterium to the genus level: (1) confirmed levan production, (2) confirmed pectinolytic activity, (3) confirmed ability to grow at 40°C, (4) inability to grow under anaerobic conditions, and (5) a negative oxidase test (Schaad et al. 2000). All test results identified the genus as Burkholderia. To identify to species level, gyrase subunit B (gyrB) and RNA polymerase subunit D (rpoD) markers were PCR amplified and sequenced using primers UP1-E/AprU, and 70F2/70R2, respectively (Maeda et al. 2006). NCBI GenBank BLASTn comparison showed that the gyrB sequence shared 99.33% identity to the type strain of B. gladioli (CP009323.1), while the rpoD sequence showed 99.53% identity (CP009322.1). Sequences were deposited in GenBank under accession numbers ON597852 (gyrB) and ON597853 (rpoD). To confirm pathogenicity, each of two Gladiolus communis 'Mini Elvira' potted plants were inoculated with two bacterial and two control treatments (3 leaves/treatment/plant) as follows: leaf infiltration with 1 mL of either (i) a distilled water-Tween 20 (0.03% v/v) bacterial suspension (106 cfu/mL) or (ii) a sterile water-Tween 20 suspension using a needle-less syringe; foliar spray with either (iii) the bacterial suspension or (iv) water-Tween suspension until run-off. Following inoculation, plants were covered for 24 hours with a plastic bag to increase humidity and favor infection and maintained in a greenhouse at an average temperature of 23°C. After 3 days, water-soaked, necrotic lesions were observed on the inoculated plants regardless of inoculation method, while control leaves remained asymptomatic. To fulfill Koch's postulates, bacteria were re-isolated from the lesions 7 days post-inoculation and confirmed to be identical to the original isolate based on rpoD gene sequencing. Bacterial scab of gladiolus was reported in Ohio in the late 1900s as caused by Pseudomonas gladioli (syn. P. marginata; Ellett, 1989). To the best of our knowledge, this report represents the first molecular identification of the causal agent as Burkholderia gladioli. In Ohio, the pathogen has also been observed causing slippery skin on onion but not officially reported in the peer-reviewed literature. Additionally, B. gladioli has been reported in other parts of the United States on orchid, corn, and rice (Keith et al. 2005; Lu et al. 2007; Nandakumar et al. 2009). Given the significant role of gladiolus within Ohio's floricultural trade, as well as the ability of this pathogen to infect other regional crops, monitoring of bacterial scab is important for floriculture and field crop growers alike.

Comparison of point-of-care glucometers and laboratory based glucose oxidase test in determining blood glucose levels

Alterations in blood glucose levels are common and an important determinant of a patient's admission outcomes, point-of-care glucometers, which are affected by a variety of factors, are increasingly used in clinical care. In this study we compared blood glucose levels determined by two commonly used glucometers (One Touch® and Accu-check®) with those of a standard laboratory method and determined the effect of haematocrit on glucose readings. Blood glucose levels were measured with One Touch® and Accu-Check® glucometers and the glucose oxidase method at the same time in 295 children aged 0 to 15 years over a 6-month period. Bland-Altman and correlation analysis were used to explore biases among the three methods. For all statistical tests, a p-value of less than 0.05 was considered statistically significant. Most were males (51.2%) and the median (range) age was 1 year (1 day, 12 years). There was a significant correlation between each of the glucometer methods and laboratory blood sugar, and the correlation between the two glucometers was strong and significant. This correlation remained statistically significant even after controlling for haematocrit values. There was an acceptable level of bias (3.9 mg/dL) between the One Touch® and Accu-check® glucometers, but each had a remarkably large bias compared with the glucose oxidase method. The use of a tested glucometer in clinical settings can aid in rapid decision-making, but there is a need to periodically cross-check with the glucose oxidase method in the laboratory to optimise treatment outcomes for children with dysglycaemia.

Niabella agricola sp. nov., isolated from paddy soil.

A polyphasic taxonomic approach was used to characterize a Gram-stain-negative, orange-coloured bacterium (designated strain CC-SYL272T) isolated from paddy soil. Cells were observed to be strictly aerobic, non-motile and non-spore-forming rods, exhibiting positive catalase and oxidase. Strain CC-SYL272T was found to grow optimally at 20-40 °C, pH 6.0-8.0 and NaCl 0-2 % (w/v). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain CC-SYL272T belongs to the genus Niabella, family Chitinophagaceae, and is most closely related to Niabella pedocola (97.8 %) followed by Niabella drilacis (97.2 %) and established a distinct taxonomic lineage associated with these species. The highest orthologous average nucleotide identity (OrthoANI) values were recorded for strain CC-SYL272T versus Niabella species (69.1-83.5 %, n=8). The mean digital DNA-DNA hybridization (dDDH) value obtained for strain CC-SYL272T against N. pedocola was 27.3 %. The polar lipid profile consisted of phosphatidylethanolamine and five unidentified lipids. The major polyamines were putrescine and sym-homospermidine. The dominating cellular fatty acids (>5 %) included iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3OH and C16 : 1 ω6c/C16 : 1 ω7c. The draft genome (6.25 Mb) of strain CC-SYL272T spanned three contigs having 47.1 mol% DNA G+C content, 5087 protein-encoding genes, 10 rRNA genes and 44 tRNA genes. The genome harboured genes involved in the depolymerization of both animal and plant polysaccharides. Based on its distinct phylogenetic, phenotypic and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence, OrthoANI, dDDH and the phylogenomic placement, strain CC-SYL272T is considered to represent a novel species of the genus Niabella, affiliated to the family Chitinophagaceae, for which the name Niabella agricola sp. nov. is proposed. The type strain is CC-SYL272T (=BCRC 81319T=JCM 34758T).

Pseudomonas oryzagri sp. nov., isolated from a rice field soil.

A Gram-stain-negative, aerobic, rod-shaped and non-motile novel bacterial strain, designated MAHUQ-58T, was isolated from soil sample of a rice field. The colonies were observed to be light pink-coloured, smooth, spherical and 0.6-1.0 mm in diameter when grown on nutrient agar (NA) medium for 2 days. Strain MAHUQ-58T was found to be able to grow at 15-40 °C, at pH 5.5-10.0 and with 0-1.0 % NaCl (w/v). Cell growth occurred on tryptone soya agar, Luria-Bertani agar, NA, MacConkey agar and Reasoner's 2A agar. The strain was found to be positive for both oxidase and catalase tests. The strain was positive for hydrolysis of Tween 20 and l-tyrosine. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Pseudomonas and to be closely related to Pseudomonas oryzae WM-3T (98.9 % similarity), Pseudomonas linyingensis LYBRD3-7T (97.7 %), Pseudomonas sagittaria JCM 18195 T (97.6 %) and Pseudomonas guangdongensis SgZ-6T (97.2 %). The novel strain MAHUQ-58T has a draft genome size of 4 536 129 bp (46 contigs), annotated with 4064 protein-coding genes, 60 tRNA genes and four rRNA genes. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain MAHUQ-58T and four closely related type strains were in the range of 85.5-89.5 % and 29.5-38.0 %, respectively. The genomic DNA G+C content was determined to be 67.0 mol%. The predominant isoprenoid quinone was ubiquinone 9. The major fatty acids were identified as C16:0, summed feature 3 (C16 : 1 ω6c and/or C16 : 1 ω7c) and summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c). On the basis of dDDH and ANI values, genotypic results, and chemotaxonomic and physiological data, strain MAHUQ-58T represents a novel species within the genus Pseudomonas, for which the name Pseudomonas oryzagri sp. nov. is proposed, with MAHUQ-58T (=KACC 22005T=CGMCC 1.18518T) as the type strain.

COMPARATIVE ANALYSIS OF URINARY TRACT INFECTIONS AMONG UNIVERSITY STUDENTS, BOYS AND GIRLS

Urinary tract infection (UTI) represents a serious health problem. The severity of urinary tract infection depends both on the virulence of the bacteria and the susceptibility of the host. The aim of this study was to determine the prevalence of urinary tract infection and demographic differentials among students, and also to treat effectively the infection, prevent urosepsis and reduce the likelihood of renal damage and provide symptomatic relief to patients. The objectives were to determine the proportion of students who have UTI, identify the common causative organism and to determine difference in UTI occurrence among students based on demographic variables. Ten students within the period of study were investigated. Personal data were collected using a structured questionnaire. Early morning midstream clean urine samples were collected from all participants who were willing to participate for analysis of UTI. Urine samples were cultured using standard loop technique on blood agar, cystein lactose electrolyte deficient medium agar and macconkey agar. Gram staining and biochemical test. After subculture the isolates were both Gram negative and positive, catalase test positive, oxidase test negative, citrate test negative, coagulase test positive, indole test negative. Total colony counts in the samples from the girls 68 105, 99 105, 21 103, 0 1 , 34 5. Then from the boys were 14 2 21 104, 18 103, 14 102, 0 101. Sensitivity test were carried out using Kirby-bauer method (disck diffusion method). The organisms were tested with antibiotics disc. . The tests were carried out on Mueller-Hinton agar, and incubated for 37degree celcius for 24hrs. All the bacteria were screened against Nitrofurantoin ceftriaxone, ciprofloxacin, gentamicin, ofloxacin, augmentin, pefloxacin, clarithromycin, chloramphenicol, ampicillin,. three ( 3 ) out of all the antibiotics bacteria that was isolated were sensitive to Nitrofurantoin, ampicillin, ciprofloxacin, While 5 were resistant to the other five (5) antibiotics , ceftriaxone, gentamicin, ofloxacin, augumentin, pefloxacin, clarithromycin, chloramphenicol. Nitrofuration, ciprofloxacinand Ampicilin showed the highest zone of inhibition in the urine sample analysis .by calculating the mean of inhibition zone diameters in replicate plates. Conclusion the general results show that Screening for UTI is important in every students. Based on the results Nitrofurantoin and ciprofloxacin, and ampicillin are the drug of the choice due to high resistant rate to the commonly used antibiotic we highly recommend it since is the most sensitive antibiotic and should be used.

Occurrence and Antimicrobial Susceptibility Profile of Escherichia Coli Isolates from Vended Milk in University of Abuja Community, Federal Capital Territory, Nigeria.

Purpose: This study was conducted to establish occurrence and antimicrobial susceptibility profile of Escherichia coli (E. coli) isolates from cow milk (Nono) sold within University of Abuja campuses.&#x0D; Methodology: Fifty (50) samples of “nono” milk were randomly collected from vendors in mini and main campus of University of Abuja from August to October 2021. All samples were analyzed using pour and surface plating methods. Presumptive E. coli isolates on Eosin methylene blue agar were characterized using preliminary and standard biochemical identification methods such as microbial plate count, Gram staining, oxidase, indole, methyl red, voges-proskauer, citrate and catalase tests. Antimicrobial susceptibility testing was conducted on positive isolates using the disc diffusion method.&#x0D; Findings: Out of the 50 samples of nono milk cultured, thirty (30) yielded growth for Escherichia coli which appeared as greenish metallic sheen on EMB agar plate and Gram-negative short rods on microscopic examination, representing an overall prevalence of 60%. The overall viable count of all samples was extremely high, with 8.7x10^7cfu/ml recorded as the highest colony forming unit per milliliter (CFU/ml) and 1.5×10^7cfu/ml recorded the lowest colony forming unit per milliliter (CFU/ml). All isolates were indole positive, methyl red positive, oxidase negative, citrate negative, catalase positive and voges-proskauer negative. E. coli isolates in this study were susceptible to sparfloxacin (66.7%), gentamicin (66.7%), ciprofloxacin (60%), streptomycin (60%), and tarivid (53.3%) and resistant to Septrin (73.3%), Augumentin (60%), Pefloxacin (66.7%), chloramphenicol (60%) and amoxicillin (53.3%). In conclusion, this study showed that the degree of contamination of E. coli in vended milk in the study area is higher than the maximum permissible bacteria count recommended by Codex Alimentarius standard for fermented milk products which is indicative of potential hazard to consumers.&#x0D; Recommendation: Public enlightenment on hygienic handling of milk during milking and especially vending by milk vendors is hereby recommended. Use of cool thermos for milk hawking is also advocated.