Molecular analysis of tetracycline resistant gene in gram-negative bacteria isolated from dairy farms

2020 ◽  
Keyword(s):  
Klebsiella Pneumonia ◽  
Gram Negative Bacteria ◽  
Aseptic Technique ◽  
Bacterial Dna ◽  
Gram Negative ◽  
Resistant Gene ◽  
Gram Staining ◽  
Oxidase Test ◽  
Thermal Cycler ◽  
Klebsiella Spp

The present study was conducted from April 2019 to June 2019 in order to detect the tetracycline A resistant Gene in gram negative bacteria. A total of 40 buffalo's milk samples were collected randomly by aseptic technique, brought to laboratory. They were inoculated on Blood and MacConkey agars and then incubated at 37°C for 24 hours whereby growth of colonies were further confirmed with catalase test, Coagulase test, Oxidase test, Gram staining and API 20 E kit. Bacterial DNA was isolated using the boiling method. The Tet A gene (210 bp) was amplified in thermal cycler and run on 1.8-gram agarose gel with 50 kb ladder. The most predominant bacterial colonies observed were of Escherichia coli (10 (33.3%) followed by Klebsiella pneumonia 5 (16.7%), Klebsiella spp. 5 (16.7 %), Pseudomonas spp. 10 (33.3%) and prevalence of tetracycline A gene was 8 (26.7%).

scholarly journals Klebsiella pneumoniae Isolated from Subclinical Mastitis Milk of Etawah Crossbreed Goat

2019 ◽  
Vol 36 (2) ◽  
pp. 238
Author(s):  
Clara Ajeng Artdita ◽  
Fajar Budi Lestari ◽  
Achmad Fauzi ◽  
Erian Pemila Ayu Tanzila
Keyword(s):  
Klebsiella Pneumoniae ◽  
Dairy Farm ◽  
Subclinical Mastitis ◽  
Positive Control ◽  
Gram Negative Bacteria ◽  
Biochemical Tests ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Staining ◽  
Klebsiella Spp

Klebsiella spp. is a common bacteria causing mastitis. Klebsiella pneumoniae and Klebsiella varicola is the most infected Klebsiella spp in the dairy farm. This study used 16 milk samples from 8 lactation goats in Sahabat Ternak farm, Sleman, Yogyakarta. Samples were tested by california mastitis test (CMT) reagent to determine the mastitis status, inoculated on blood agar (BA) then on Mac Conkey Agar (MCA) as a selective media for Gram negative bacteria. The colonies from BA and MCA were stained by Gram staining to determine Gram-positive or Gram-negative bacteria and its cell morphology. The colonies from MCA were identified by biochemical tests such as sugar fermentation tests (glucose, lactose, maltose, and saccharose), arginine, ornithine decarboxylase, indole, citrate, urea hydrolysis (Christensen’s method), lysin and malonate. The result of mastitis test showed 5 goats were positive result and 3 samples were negative. Bacterial staining showed 2 samples were Gram-negative, rods and the others were Gram-positive, coccus. The samples with rods shapes were continued by biochemical tests. The characterization result of biochemical test indicated that the rods shapes bacteria were Klebsiella pneumoniae. These bacteria identification conducted in Laboratorium Preklinis Program Studi Kesehatan Hewan, Sekolah Vokasi Universitas Gadjah Mada and the result confirmation using positive control of  Klebsiella pneumoniae conducted in Balai Laboratorium Kesehatan Yogyakarta. These results showed that milk from Etawah crossbreed goat was infected by Klebsiella pneumoniae.

Dual stain kit for the microscopic gram classification of ocular infection bacteria

1993 ◽  
Vol 51 ◽  
pp. 248-249
Author(s):  
Jacob S. Hanker ◽  
Dale N. Holdren ◽  
Kenneth L. Cohen ◽  
Beverly L. Giammara
Keyword(s):  
Contact Lenses ◽  
Ocular Infection ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Gram Staining ◽  
Ocular Infections ◽  
Different Types ◽  
Culture Positive ◽  
Increased Susceptibility

Keratitis and conjunctivitis (infections of the cornea or conjunctiva) are ocular infections caused by various bacteria, fungi, viruses or parasites; bacteria, however, are usually prominent. Systemic conditions such as alcoholism, diabetes, debilitating disease, AIDS and immunosuppressive therapy can lead to increased susceptibility but trauma and contact lens use are very important factors. Gram-negative bacteria are most frequently cultured in these situations and Pseudomonas aeruginosa is most usually isolated from culture-positive ulcers of patients using contact lenses. Smears for staining can be obtained with a special swab or spatula and Gram staining frequently guides choice of a therapeutic rinse prior to the report of the culture results upon which specific antibiotic therapy is based. In some cases staining of the direct smear may be diagnostic in situations where the culture will not grow. In these cases different types of stains occasionally assist in guiding therapy.

Quality Improvement of the DNA extracted by boiling method in Gram negative bacteria

2017 ◽  
Vol 6 (04) ◽  
pp. 5347 ◽  
Author(s):  
Omar B. Ahmed* ◽  
Anas S. Dablool
Keyword(s):  
Dna Extraction ◽  
Control Method ◽  
Extraction Procedure ◽  
Cost Effective ◽  
High Yield ◽  
Gram Negative Bacteria ◽  
Bacterial Dna ◽  
Gram Negative ◽  
E Coli

Several methods of Deoxyribonucleic acid (DNA) extraction have been applied to extract bacterial DNA. The amount and the quality of the DNA obtained for each one of those methods are variable. The study aimed to evaluate bacterial DNA extraction using conventional boiling method followed by alcohol precipitation. DNA extraction from Gram negative bacilli was extracted and precipitated using boiling method with further precipitation by ethanol. The extraction procedure performed using the boiling method resulted in high DNA yields for both E. coli and K. pneumoniae bacteria in (199.7 and 285.7μg/ml, respectively) which was close to control method (229.3 and 440.3μg/ml). It was concluded that after alcohol precipitation boiling procedure was easy, cost-effective, and applicable for high-yield quality of DNA in Gram-negative bacteria.

scholarly journals Pyelonephritis in Pediatric Uropathic Patients: Differences from Community-Acquired Ones and Therapeutic Protocol Considerations. A 10-Year Single-Center Retrospective Study

Children ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 436
Author(s):  
Giovanni Parente ◽  
Tommaso Gargano ◽  
Stefania Pavia ◽  
Chiara Cordola ◽  
Marzia Vastano ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Retrospective Study ◽  
Success Rate ◽  
Pediatric Population ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Significant Difference ◽  
Klebsiella Spp

Pyelonephritis (PN) represents an important cause of morbidity in the pediatric population, especially in uropathic patients. The aim of the study is to demonstrate differences between PNs of uropathic patients and PNs acquired in community in terms of uropathogens involved and antibiotic sensitivity; moreover, to identify a proper empiric therapeutic strategy. A retrospective study was conducted on antibiograms on urine cultures from PNs in vesicoureteral reflux (VUR) patients admitted to pediatric surgery department and from PNs in not VUR patients admitted to Pediatric Emergency Unit between 2010 and 2020. We recorded 58 PNs in 33 patients affected by VUR and 112 PNs in the not VUR group. The mean age of not VUR patients at the PN episode was 1.3 ± 2.6 years (range: 20 days of life–3 years), and almost all the urine cultures, 111 (99.1%), isolated Gram-negative bacteria and rarely, 1 (0.9%), Gram-positive bacteria. The Gram-negative uropathogens isolated were Escherichia coli (97%), Proteus mirabilis (2%), and Klebsiella spp. (1%). The only Gram-positive bacteria isolated was an Enterococcus faecalis. As regards the antibiograms, 96% of not VUR PNs responded to beta-lactams, 99% to aminoglycosides, and 80% to sulfonamides. For the VUR group, mean age was 3.0 years ± 3.0 years (range: 9 days of life–11 years) and mean number of episodes per patient was 2.0 ± 1.0 (range: 1–5); 83% of PNs were by Gram-negatives bacteria vs. 17% by Gram-positive: the most important Gram-negative bacteria were Pseudomonas aeruginosa (44%), Escherichia coli (27%), and Klebsiella spp. (12%), while Enterococcus spp. determined 90% of Gram-positive UTIs. Regimen ampicillin/ceftazidime (success rate: 72.0%) was compared to ampicillin/amikacin (success rate of 83.0%): no statistically significant difference was found (p = 0.09). The pathogens of PNs in uropathic patients are different from those of community-acquired PNs, and clinicians should be aware of their peculiar antibiotic susceptibility. An empiric therapy based on the association ampicillin + ceftazidime is therefore suggested.

scholarly journals Utilization of Vector Autoregressive and Linear Transfer Models to Follow Up the Antibiotic Resistance Spiral in Gram-negative Bacteria From Cephalosporin Consumption to Colistin Resistance

2018 ◽  
Vol 69 (8) ◽  
pp. 1410-1421 ◽  
Author(s):  
Hajnalka Tóth ◽  
Adina Fésűs ◽  
Orsolya Kungler-Gorácz ◽  
Bence Balázs ◽  
László Majoros ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Carbapenem Resistance ◽  
Gram Negative Bacteria ◽  
Vicious Cycle ◽  
Var Models ◽  
Gram Negative ◽  
Vector Autoregressive ◽  
E Coli ◽  
Cephalosporin Resistance ◽  
Klebsiella Spp

Abstract Background Increasing antibiotic resistance may reciprocally affect consumption and lead to use of broader-spectrum alternatives; a vicious cycle that may gradually limit therapeutic options. Our aim in this study was to demonstrate this vicious cycle in gram-negative bacteria and show the utility of vector autoregressive (VAR) models for time-series analysis in explanatory and dependent roles simultaneously. Methods Monthly drug consumption data in defined daily doses per 100 bed-days and incidence densities of gram-negative bacteria (Escherichia coli, Klebsiella spp., Pseudomonas aeruginosa, and Acinetobacter baumannii) resistant to cephalosporins or to carbapenems were analyzed using VAR models. These were compared to linear transfer models used earlier. Results In case of all gram-negative bacteria, cephalosporin consumption led to increasing cephalosporin resistance, which provoked carbapenem use and consequent carbapenem resistance and finally increased colistin consumption, exemplifying the vicious cycle. Different species were involved in different ways. For example, cephalosporin-resistant Klebsiella spp. provoked carbapenem use less than E. coli, and the association between carbapenem resistance of P. aeruginosa and colistin use was weaker than that of A. baumannii. Colistin use led to decreased carbapenem use and decreased carbapenem resistance of P. aeruginosa but not of A. baumannii. Conclusions VAR models allow analysis of consumption and resistance series in a bidirectional manner. The reconstructed resistance spiral involved cephalosporin use augmenting cephalosporin resistance primarily in E. coli. This led to increased carbapenem use, provoking spread of carbapenem-resistant A. baumannii and consequent colistin use. Emergence of panresistance is fueled by such antibiotic-resistance spirals.

scholarly journals ISOLASI DAN PENENTUAN INDEKSHIDROLISIS BAKTERI AMILOLITIK DARI TANAH SEDIMENT MANGROVE DI WONOREJO, SURABAYA

2019 ◽  
Vol 10 (1) ◽  
pp. 38
Author(s):  
Aliyah Siti Sundari ◽  
Ni Nyoman Purwani ◽  
Anita Kurniati
Keyword(s):  
Soil Bacteria ◽  
Mangrove Ecosystem ◽  
Soil Characteristics ◽  
Mangrove Sediment ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Lugol’S Iodine ◽  
Gram Staining ◽  
Special Soil

Mangrove sediment is a habitat for various bacteria, one of them is amylolytic bacteria which has the potential to produce amylase enzyme. Amylase enzyme has many benefits in industry, textiles and medical. The mangrove ecosystem area has special soil characteristics, which have the opportunity to have microorganism diversity, one of which is the mangrove ecosystem in the Wonorejo region, Surabaya. This study aims to obtain potential amylolytic bacteria from potential amylolytic isolates derived from mangrove sediment. Soil bacteria were isolated in Starch media for 2% agar and tested with Lugol’s Iodine reagents to measure their amylolytic index. Character isolates observed included colony morphology, Gram staining, and motility. Of the 27 isolates found there were 3 isolates with the highest index values in their activity, namely isolates A.7, A.27 and A.64. Characterization results showed that isolates A.7 and A.64 were Gram negative bacteria, and isolates A.27 were Gram positive bacteria. And the motility results for the three isolates were negative, with the results of a positive catalase test.

Select Gram-negative Aerobic Bacteria

2012 ◽  
pp. 90-101
Author(s):  
David R. McNamara ◽  
Franklin R. Cockerill
Keyword(s):  
Cell Wall ◽  
Sepsis Syndrome ◽  
Vital Role ◽  
Klebsiella Pneumonia ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Neisseria Meningitides ◽  
A Cell ◽  
Normal Human ◽  
Specific Species

Gram-negative bacteria may be rod-shaped (bacilli), spherical (cocci), oval, helical, or filamentous. Cytoplasmic membrane is surrounded by a cell wall consisting of a peptidoglycan layer and an outer cell membrane. Gram-negative bacteria are widely distributed in the natural environment. They are commensals with many animals and play a vital role in normal human physiology as intestinal commensals. Gram-negative bacteria are the cause of various human illnesses. The gram-negative bacterial cell wall contains various lipopolysaccharide endotoxins. Endotoxins trigger intense inflammation and the sepsis syndrome during infection. Specific species of gram-negative bacteria such as Neisseria meningitides, Moraxella catarrhalis, Acinetobacter, Vibrio, Klebsiella pneumonia, Salmonella, Pseudomonas aeruginosa, and Haemophilus influenza are reviewed.

Susceptibility of Gram-negative bacteria to the synergistic bactericidal action of serum and polymyxin B nonapeptide

1986 ◽  
Vol 32 (1) ◽  
pp. 66-69 ◽  
Author(s):  
Petri Viljanen ◽  
Helena Käyhty ◽  
Martti Vaara ◽  
Timo Vaara
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Enterobacter Cloacae ◽  
Polymyxin B ◽  
Gram Negative Bacteria ◽  
Bactericidal Action ◽  
Gram Negative ◽  
Degree Of Sensitization ◽  
Normal Human ◽  
Klebsiella Spp

Polymyxin B nonapeptide was able to sensitize Escherichia coli strains and strains of Salmonella typhimurium, Klebsiella spp., Enterobacter cloacae, Pseudomonas aeruginosa, and Haemophilus influenzae to the bactericidal action of fresh normal human serum. The degree of sensitization varied significantly within the strains. Strains of Proteus mirabilis, Neisseria gonorrhoeae, and N. meningitidis remained resistant.

Species distribution and antimicrobial susceptibility profile of gram-negative bacteria in hospitalized cancer patients

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e20730-e20730
Author(s):  
H. M. Ashour ◽  
A. El-Sharif
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Cancer Patients ◽  
Solid Tumor ◽  
Klebsiella Pneumonia ◽  
Gram Negative Bacteria ◽  
Isolation And Identification ◽  
Tumor Patients ◽  
Acinetobacter Species ◽  
Gram Negative

e20730 Background: Nosocomial infections pose significant threats to hospitalized patients, especially the immunocompromised ones, such as cancer patients. Methods: This study examined the microbial spectrum of gram-negative bacteria in various infection sites in patients with leukemia and solid tumors. The antimicrobial resistance patterns of the isolated bacteria were studied. Results: The most frequently isolated gram-negative bacteria were Klebsiella pneumonia (31.2%) followed by Escherichia coli (22.2%). We report the first-time isolation and identification of a number of less-frequent gram negative bacteria (Chromobacterium violacum, Burkholderia cepacia, Kluyvera ascorbata, Stenotrophomonas maltophilia, Yersinia pseudotuberculosis, and Salmonella arizona). Most of the gram-negative isolates from RTI, GITI, UTI, and BSI were obtained from leukemic patients. All gram-negative isolates from SI were obtained from solid-tumor patients. In both leukemic and solid-tumor patients, gram-negative bacteria causing UTI were mainly Escherichia coli and Klebsiella pneumoniae, while gram-negative bacteria causing RTI were mainly Klebsiella pneumoniae. Escherichia coli was the main gram-negative pathogen causing BSI in solid-tumor patients and GITI in leukemic patients. Isolates of Escherichia coli, Klebsiella, Enterobacter, Pseudomonas, and Acinetobacter species were resistant to most antibiotics tested. There was significant imipenem-resistance in Acinetobacter (40.9%), Pseudomonas (40%), and Enterobacter (22.2%) species, and noticeable imipinem-resistance in Klebsiella (13.9%) and Escherichia coli (8%). Conclusions: This is the first study to report the evolution of imipenem-resistant gram-negative strains in Egypt. Mortality rates were higher in cancer patients with nosocomial Pseudomonas infections than any other bacterial infections. Policies restricting antibiotic consumption should be implemented to avoid the evolution of newer generations of antibiotic resistant-pathogens. No significant financial relationships to disclose.

scholarly journals A Physical-Chemical Study of the Interference of Ceftriaxone Antibiotic with Copper Chloride Salt

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Elsayed M. AbouElleef ◽  
Mowafak M. Mahrouka ◽  
Sherine E. Salem
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Copper Chloride ◽  
Chemical Study ◽  
Antimicrobial Activities ◽  
Klebsiella Pneumonia ◽  
Chloride Salt ◽  
Gram Negative Bacteria ◽  
Zone Of Inhibition ◽  
Conductivity Method ◽  
Gram Negative

The nano-CuCl2.2H2O salt was prepared by the ball milling method. The association parameters for bulk and nano-CuCl2 salts in H2O are estimated at different temperatures using the conductivity method by applying the Fuoss–Shedlovsky equation and it was higher for nano-CuCl2 than bulk CuCl2 salt. The interaction between the cation (Cu2+) and ligand (ceftriaxone) in H2O was determined also by the conductometric method. Two stoichiometric complexes 1/2 and 1/1 (M/L) are estimated and follow the order Kf (1/1) > Kf (1 : 2) and ∆Gf (1/1) > ∆Gf (1/2) for (M : L) (in negative values) indicate the favorable of formation of (1/1) complex compared to the (1 : 2) complex. The Gibbs free energies change was increased in negative signs with increasing the temperature. The antimicrobial activities of CFT, bulk Cu-CFT complex, and nano-Cu-CFT complex were studied on LB agar by the disc diffusion technique against clinical isolates of gram-negative bacteria (Klebsiella pneumonia and Pseudomonas aeruginosa) and Fungi (Candida albicans). It was observed that (CFT) has a higher zone of inhibition and antibacterial activity than that of bulk and nano-Cu-CFT complexes in Klebsiella pneumonia and Pseudomonas aeruginosa (gram-negative bacteria). The nano-Cu-CFT complex showed a higher clear zone of inhibition and antifungal activity against candida than bulk Cu-CFT complex while the absence of the inhibition zone in CFT, so nano-Cu-CFT complex, can be used as an antifungal drug.

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