scholarly journals Isolation and Identification of Bacteria Responsible for the Spoilage of Fluted Pumpkin (Telfaria occidentalis) and Bitter Leaf (Vernonia amygdalina) in Sokoto Metropolis

Author(s):  
A.S. Muhammad ◽  
S.A. Ani ◽  
A.M. Nasiru

The research was conducted in 2016 at the Microbiology Laboratory in the Department of Microbiology in Usmanu Danfodiyo University Sokoto. The main objective was to isolate and identify those bacteria responsible for the spoilage of bitter leaf (Vernonia amygdalina) and fluted pumpkin (Telfairia occidentalis). This approach presented extracting these bacteria form the vegetables and then culture then on agar plates, colonies were formed and subcultures on different plates, pure cultures were obtained and cultured in slant bottles containing nutrient agar for identification through biochemical test bacterial utilization of substrates which include Triple sugar ion, Methyl red and Vogues prosturer test, starch hydrolysis, Indole, citrate, catalase, urease test and recorded. Gram stain was carried out to identify the gram reaction and viewed under microscope for cell morphology. Bacteria species of the stock culture were identified using Atlas Book of Bacteria using the biochemical test results and gram stain reactions, the bacteria isolated were identified as gram negative rod bacteria. Percentage distribution of occurrence showed that E. coli had had the highest frequency of occurrence with percentage distribution of 42.8% in bitter leaf, followed by Proteus mirabilis, and B. cereus both having 28.6% respectively. Proteus mirabilis had 50% in fluted pumpkin, Klebsiella 33.3%, E. coli 16.7%. From the result, it was concluded that E. coli and Proteus had the highest occurrence of bacterial contamination of spoilt bitter leaf and fluted pumpkin sold in Sokoto metropolis and recommended sensitization proper hygienic measure during post-harvest handling of these vegetables. KEYWORDS: Isolation, Identification, Bacteria, Vegetables, Food Spoilage, Hygiene

2016 ◽  
Vol 79 (10) ◽  
pp. 1789-1793 ◽  
Author(s):  
HIDEMI IZUMI ◽  
YUJI NAKATA ◽  
AYANO INOUE

ABSTRACT Chemical sanitizers may induce no injury (bacteria survive), sublethal injury (bacteria are injured), or lethal injury (bacteria die). The proportion of coliform bacteria that were injured sublethally by chlorine and fungicide mixed with agricultural water (pond water), which was used to dilute the pesticide solution, was evaluated using the thin agar layer (TAL) method. In pure cultures of Enterobacter cloacae, Escherichia coli, and E. coli O157:H7 (representing a human pathogen), the percentage of chlorine-injured cells was 69 to 77% for dilute electrolyzed water containing an available chlorine level of 2 ppm. When agricultural water was mixed with electrolyzed water, the percentage of injured coliforms in agricultural water was 75%. The isolation and identification of bacteria on TAL and selective media suggested that the chlorine stress caused injury to Enterobacter kobei. Of the four fungicide products tested, diluted to their recommended concentrations, Topsin-M, Sumilex, and Oxirane caused injury to coliform bacteria in pure cultures and in agricultural water following their mixture with each pesticide, whereas Streptomycin did not induce any injury to the bacteria. The percentage of injury was 45 to 97% for Topsin-M, 80 to 87% for Sumilex, and 50 to 97% for Oxirane. A comparison of the coliforms isolated from the pesticide solutions and then grown on either TAL or selective media indicated the possibility of fungicide-injured Rahnella aquatilis, Yersinia mollaretii, and E. coli. These results suggest the importance of selecting a suitable sanitizer and the necessity of adjusting the sanitizer concentration to a level that will kill the coliforms rather than cause sanitizer-induced cell injury that can result in the recovery of the coliforms.


2011 ◽  
Vol 74 (2) ◽  
pp. 221-227 ◽  
Author(s):  
LUISA Y. SOLÍS-SOTO ◽  
SANTOS GARCÍA ◽  
IRENE WESLEY ◽  
NORMA HEREDIA

The microaerophilic nature of Campylobacter and its requirement of ~5% O2 for growth have complicated its recovery from foods. The addition to the enrichment media of oxygen quenchers such as charcoal or blood could interfere with PCR for its detection. In this study, a two-step simple aerobic method for Campylobacter detection is proposed. A modification of the Tran blood-free enrichment broth (BFEB), in which charcoal was excluded from the medium (M-BFEB), was compared with the original formulation and other enrichment broths. Campylobacter jejuni and Campylobacter coli were screened by PCR directly from the enrichment media. Various levels of pure cultures of C. jejuni and C. coli combined with Escherichia coli were inoculated into Preston, Bolton, BFEB, and the modified BFEB (M-BFEB). In addition, Campylobacter was inoculated onto retail purchased chicken skin and recovery was quantified. Rates of recovery after 24 to 48 h of enrichment at 42°C under aerobic incubation for BFEB and M-BFEB and microaerobic incubations for Preston and Bolton broths were determined. Overall, our results indicated that the most sensitive medium was Bolton's, followed by either BFEB or M-BFEB; the least sensitive was Preston's. M-BFEB was directly coupled to a PCR assay to detect Campylobacter, avoiding intermediate plating. Campylobacter was detected in the presence of up to108 E. coli cells per ml. M-BFEB facilitated detection of both C. jejuni and C. coli artificially inoculated onto chicken skin samples. M-BFEB coupled to PCR is a rapid and attractive alternative for isolation and identification of C. coli and C. jejuni from poultry.


2019 ◽  
Vol 5 (1) ◽  
pp. 38-44
Author(s):  
Erni Yohani Mahtuti ◽  
Farahdita Devi Masyitoh

Laboratory waste produced unique characteristics, contrast to waste produced by industrial activities. Material waste that comes from the laboratory has greater variety of waste types; although the amount of material discarded is not many. The research objective was to obtain bacterial isolates that were able to survive in laboratory waste as potential waste-degrading bacteria. Research method is observartional laboratory with isolate reaction testing that was detected by the ability to degrade starch, cellulose, proteins and non-organic compounds. The sampling method was purposive sampling. The stages in this study were divided into two; first, the manufacture of pure cultures from the inoculants previously diluted, then microscopic observations. The second, identification and biochemical test according to Bergey's Manual of Bacteriology Determination. Bacteria were rejuvenated on medium nutrient so that the isolates were obtained twenty four hours old. Then an examination was carried out include Gram staining. Enzymatic test of amylase, protease, and cellulose, and biochemical test to identify microbes that degrade chemical compounds includes; oxidase test, motility, nitrate, lysine, ornithine, H2O, Glucose, Mannitol, xylose, ONPG, Indole, urease, V-P, citrate, TDA. The results of the study were found Pseudomonas stutzeri, Proteus mirabilis, and Pseudomonas aeruginosa. Isolates that have an amylolytic index are C1, C2 and O7 namely Pseudomonas stutzeri, Proteus mirabilis, and Pseudomonas aeruginosa. The resulting index was C1 = 0.45, C2 = 0.65 and O7 = 0.87. Keywords: Isolation, identification, laboratory liquid waste, waste degradation microbes ABSTRAK Limbah laboratorium menghasilkan karakteristik yang unik, kontras dengan limbah yang dihasilkan oleh kegiatan industri. Limbah bahan yang berasal dari laboratorium memiliki jenis sampah yang lebih banyak, meskipun jumlah bahan yang dibuang tidak banyak. Tujuan penelitian adalah untuk memperoleh isolat bakteri yang mampu bertahan hidup di dalam limbah laboratorium sebagai bakteri pengurai limbah potensial. Metode penelitian adalah observasional laboratorium dengan melakukan tes reaksi isolat untuk mengetahui kemampuan degradasi pati, selulosa, protein dan senyawa non-organik. Teknik pengambilan sampel adalah purposive sampling. Tahapan dalam penelitian ini dibagi menjadi dua yaitu pertama, pembuatan kultur murni dari inokulan yang sebelumnya diencerkan, kemudian pengamatan mikroskopis. Kedua, identifikasi dan uji biokimia sesuai dengan Manual Penentuan Bakteriologi Bergey. Bakteri diremajakan pada nutrisi sedang sehingga isolat diperoleh dalam dua puluh empat jam. Uji pemeriksaan adalah pewarnaan Gram. Selanjutnya uji enzimatik amilase, protease, dan selulosa, dan uji biokimia untuk mengidentifikasi mikroba yang mendegradasi senyawa kimia meliputi; uji oksidase, motilitas, nitrat, lisin, ornithine, H2O, Glukosa, Mannitol, xilosa, ONPG, Indole, urease, V-P, sitrat, TDA. Hasil penelitian ditemukan Pseudomonas stutzeri, Proteus mirabilis, dan Pseudomonas aeruginosa. Isolat yang memiliki indeks amilolitik adalah C1, C2 dan O7 yaitu Pseudomonas stutzeri, Proteus mirabilis, dan Pseudomonas aeruginosa. Indeks yang dihasilkan adalah C1 = 0,45, C2 = 0,65 dan O7 = 0,87. Kata kunci: Isolasi dan identifikasi mikroba, Limbah cair laboratorium, mikroba pendegradasi limbah


2012 ◽  
Vol 6 (2) ◽  
pp. 7-10
Author(s):  
Mohammad Murshed ◽  
Sabeena Shahnaz ◽  
Md. Abdul Malek

Isolation and identification of post operative hospital acquired infection was carried out from July 2008 to December 2008 in Holy Family Red Crescent Medical College Hospital (private hospital). The major pathogen of wound infection was E. coli. A total; of 120 samples were collected from the surrounding environment of post operative room like floor, bed sheets, instruments, dressing materials, catheter, nasogastric and endotracheal tube. E. coli (40%) was the predominant organism followed by S. aureus (24%). DNA fingerprinting analysis using pulsed field gel electreopheresis of XbaI restriction digested genomic DNA showed that clonal relatedness between the two clinical nd environmental isolates were 100%.DOI: http://dx.doi.org/10.3329/bjmm.v6i2.19369 Bangladesh J Med Microbiol 2012; 06(02): 7-10


1993 ◽  
Vol 27 (3-4) ◽  
pp. 267-270 ◽  
Author(s):  
M. T. Augoustinos ◽  
N. A. Grabow ◽  
B. Genthe ◽  
R. Kfir

A fluorogenic β-glucuronidase assay comprising membrane filtration followed by selective enumeration on m-FC agar at 44.5°C and further confirmation using tlie 4-metliylumbelliferyl-β-D-glucuronide (MUG) containing medium was evaluated for the detection of Escherichia coli in water. A total of 200 typical blue and non-typical blue colonies were isolated from sea and fresh water samples using initial selective enumeration on m-FC agar. Pure cultures of the selected colonies were further tested using the MUG assay and identified using the API 20E method. Of the colonies tested which were shown to be positive using the MUG assay 99.4% were Escherichia coli. The results of this study indicate the combination of the m-FC method followed by the MUG assay to be highly efficient for the selection and confirmation of E. coli from a wide range of environmental waters.


1998 ◽  
Vol 64 (12) ◽  
pp. 4862-4869 ◽  
Author(s):  
Jörg F. Rippmann ◽  
Michaela Klein ◽  
Christian Hoischen ◽  
Bodo Brocks ◽  
Wolfgang J. Rettig ◽  
...  

ABSTRACT Recently it has been demonstrated that L-form cells ofProteus mirabilis (L VI), which lack a periplasmic compartment, can be efficiently used in the production and secretion of heterologous proteins. In search of novel expression systems for recombinant antibodies, we compared levels of single-chain variable-fragment (scFv) production in Escherichia coliJM109 and P. mirabilis L VI, which express four distinct scFvs of potential clinical interest that show differences in levels of expression and in their tendencies to form aggregates upon periplasmic expression. Production of all analyzed scFvs in E. coli was limited by the severe toxic effect of the heterologous product as indicated by inhibition of culture growth and the formation of insoluble aggregates in the periplasmic space, limiting the yield of active product. In contrast, the L-form cells exhibited nearly unlimited growth under the tested production conditions for all scFvs examined. Moreover, expression experiments with P. mirabilis L VI led to scFv concentrations in the range of 40 to 200 mg per liter of culture medium (corresponding to volume yields 33- to 160-fold higher than those with E. coli JM109), depending on the expressed antibody. In a translocation inhibition experiment the secretion of the scFv constructs was shown to be an active transport coupled to the signal cleavage. We suppose that this direct release of the newly synthesized product into a large volume of the growth medium favors folding into the native active structure. The limited aggregation of scFv observed in the P. mirabilis L VI supernatant (occurring in a first-order-kinetics manner) was found to be due to intrinsic features of the scFv and not related to the expression process of the host cells. The P. mirabilis L VI supernatant was found to be advantageous for scFv purification. A two-step chromatography procedure led to homogeneous scFv with high antigen binding activity as revealed from binding experiments with eukaryotic cells.


2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S823-S823
Author(s):  
Kendra Foster ◽  
Linnea A Polgreen ◽  
Brett Faine ◽  
Philip M Polgreen

Abstract Background Urinary tract infections (UTIs) are one of the most common bacterial infections. There is a lack of large epidemiologic studies evaluating the etiologies of UTIs in the United States. This study aimed to determine the prevalence of different UTI-causing organisms and their antimicrobial susceptibility profiles among patients being treated in a hospital setting. Methods We used the Premier Healthcare Database. Patients with a primary diagnosis code of cystitis, pyelonephritis, or urinary tract infection and had a urine culture from 2009- 2018 were included in the study. Both inpatients and patients who were only treated in the emergency department (ED) were included. We calculated descriptive statistics for uropathogens and their susceptibilities. Multi-drug-resistant pathogens are defined as pathogens resistant to 3 or more antibiotics. Resistance patterns are also described for specific drug classes, like resistance to fluoroquinolones. We also evaluated antibiotic use in this patient population and how antibiotic use varied during the hospitalization. Results There were 640,285 individuals who met the inclusion criteria. Females make up 82% of the study population and 45% were age 65 or older. The most common uropathogen was Escherichia Coli (64.9%) followed by Klebsiella pneumoniae (8.3%), and Proteus mirabilis (5.7%). 22.2% of patients were infected with a multi-drug-resistant pathogen. We found that E. Coli was multi-drug resistant 23.8% of the time; Klebsiella pneumoniae was multi-drug resistant 7.4%; and Proteus mirabilis was multi-drug resistant 2.8%. The most common antibiotics prescribed were ceftriaxone, levofloxacin, and ciprofloxacin. Among patients that were prescribed ceftriaxone, 31.7% of them switched to a different antibiotic during their hospitalization. Patients that were prescribed levofloxacin and ciprofloxacin switched to a different antibiotic 42.8% and 41.5% of the time, respectively. Conclusion E. Coli showed significant multidrug resistance in this population of UTI patients that were hospitalized or treated within the ED, and antibiotic switching is common. Disclosures All Authors: No reported disclosures


PEDIATRICS ◽  
1969 ◽  
Vol 44 (4) ◽  
pp. 486-492
Author(s):  
Jerome O. Klein

Lung punctures were performed on 32 occasions in 28 infants with pneumonia to assist in specific bacteriologic diagnosis. The aspirates yielded pure cultures of Diplococcus pneumoniae and Staphylococcus aureus each in four patients and E. coli in two patients. The procedure is performed as for a thoracentesis and requires no special instruments. Three children had pneumothoraces and one had a small hemoptysis following the procedure, but only one child exhibited even minimal respiratory distress as a result of the tap. The literature on lung aspirates was reviewed with respect to the value and potential liability of the procedure. At present, diagnostic lung punctures should be considered in three groups of children with lower respiratory disease: (1) the critically ill child in whom a specific etiologic diagnosis is of major importance to guide antimicrobial therapy, (2) the child who has deteriorated while on therapy and in whom an etiologic agent is not available from the usual upper respiratory tract cultures, and (3) the child with pneumonia complicated by underlying disease or drugs limiting normal host defense mechanisms. In these three instances, the advantages of a specific etiologic diagnosis outweighs the small risk from the lung puncture.


1991 ◽  
Vol 54 (7) ◽  
pp. 496-501 ◽  
Author(s):  
ARTHUR HINTON ◽  
GEORGE E. SPATES ◽  
DONALD E. CORRIER ◽  
MICHAEL E. HUME ◽  
JOHN R. DELOACH ◽  
...  

A Veillonella species and Enterococcus durans were isolated from the cecal contents of adult broilers. Mixed cultures of Veillonella and E. durans inhibited the growth of Salmonella typhimurium and Escherichia coli 0157:H7 on media containing 2.5% lactose (w/v). The growth of S. typhimurium or E. coli 0157:H7 was not inhibited by mixed cultures containing Veillonella and E. durans on media containing only 0.25% lactose or by pure cultures of Veillonella or E. durans on media containing either 0.25% or 2.5% lactose. The mixed cultures of Veillonella and E. durans produced significantly (P<0.05) more acetic, propionic, and lactic acids in media containing 2.5% lactose than in media containing 0.25% lactose. The inhibition of the enteropathogens was related to the production of lactic acid from lactose by the E. durans and the production of acetic and propionic acids from lactic acid by the Veillonella.


2010 ◽  
Vol 10 (2) ◽  
pp. 209-215
Author(s):  
M. S. Mthembu ◽  
P. T. Biyela ◽  
T. G. Djarova ◽  
A. K. Basson

Fecal contamination of source waters and its associated intestinal pathogens continues to pose risks to public health although the extent and effect of microbial contamination of source waters gets very little attention in designing treatment plants in most developing countries. Coliform counts give an indication of the overall bacterial contamination of water and thus its safety for human consumption. However, their presence fails to provide information about the source of fecal contamination which is vital to managing fecal contamination problems in surface waters. This study explored the use of multiple antibiotic resistance (MAR) indexing as means of differentiating E. coli isolates from different sources. A total of 322 E. coli isolates were obtained from municipal wastewater and from fecal samples from domestic and wild animals. Conventional culture methods and standard chemical and biochemical tests were used for isolation and identification of E. coli. Isolates were assayed against 10 antibiotics using the micro-dilution technique. The results obtained generated antibiotic resistance profiles which were used to statistically group the isolates into different subsets. Correct source classification was obtained for 60% of human-derived and 95% non-human-derived E. coli respectively. These results indicate the validity of the usefulness of MAR indexing as a method of bacterial source tracking.


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