scholarly journals Identification and Characterization of Escherichia coli from Bronchial Plug of Broiler Chicken with Respiratory Distress

2021 ◽  
Vol 10 (10) ◽  
pp. 221-228
Author(s):  
M. H. Chhatrola D. T. Fefar ◽  
A. R. Bhadaniya B. J. Trangadia ◽  
V. A. Kalaria S. N. Ghodasara ◽  
B. B. Javia J. M. Chauhan
Keyword(s):  
Respiratory Distress ◽  
Broiler Chicken ◽  
Pcr Assay ◽  
E Coli ◽  
Methane Sulphonate ◽  
Highly Sensitive ◽  
High Prevalence ◽  
Identification And Characterization ◽  
Pooled Sample

A Twenty-five bronchial plug samples from dead broiler chicken with signs of respiratory distress collected for identification of avian pathogenic E. coli (APEC). Samples comprised of one pooled sample each from 25 poultry farms located in and around Junagadh city. Isolation, culture and colony characteristics used for primary detection followed by PCR assay targeting four virulence genes (iss, papC, tsh and vat)for confirmatory diagnosis. All bronchial plugs were found to be positive for E. coli infection by isolation. Out of 25 flock sampled, the highest number of E. coli isolates were found positive for the presence of iss gene (22) followed by tsh gene (15), vat gene (13) and papC gene (8)by PCR. On antibiogram, overall E. coli was highly sensitive to colistin (Methane sulphonate) and meropenem(100%) followed by levofloxacin (76%), ceftriaxone (64%), gentamicin (60%), amikacin (60%), co-trimoxazole (40%), amoxycilin-clavunic acid (8%) and imepenem (8%). These results indicated the high prevalence of the E. coli with variable antibiotic resistance in broiler chicken.

Development of a Multiplex PCR for Rapid Detection of Verocytotoxin-Producing Escherichia coli O26 in Raw Milk and Ground Beef

2011 ◽  
Vol 74 (1) ◽  
pp. 13-17 ◽  
Author(s):  
V. LORUSSO ◽  
A. DAMBROSIO ◽  
N. C. QUAGLIA ◽  
A. PARISI ◽  
G. LASALANDRA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Rapid Detection ◽  
Ground Beef ◽  
Raw Milk ◽  
The United States ◽  
Pcr Assay ◽  
E Coli ◽  
Identification And Characterization

Verocytotoxin-producing Escherichia coli (VTEC) O26 is an emergent pathotype that has caused an increasing number of sporadic cases and outbreaks of gastroenteritis, hemorrhagic colitis, and hemolytic uremic syndrome in the United States and Europe. Many cases are associated with the consumption of milk and undercooked or fermented meats. The stx2 strains of VTEC O26 seem to be more likely to cause human infections than isolates expressing only stx1. The isolation and identification of VTEC O26 from foods is labor intensive and time-consuming. We developed a multiplex PCR (M-PCR) assay for the identification and characterization of E. coli O26 VTEC and its detection in raw milk and ground beef. The method is based on the amplification of the wzx, stx1, and stx2 genes for the simultaneous detection of the O26 antigen and verocytotoxin types 1 and 2. This M-PCR assay had a sensitivity of 108 CFU/ml when applied to a bacterial suspension and of 106 CFU/ml or g when applied to both inoculated milk and minced beef samples. This M-PCR assay also was highly specific, and results were consistently negative for negative controls (nonpathogenic E. coli strains, uninoculated milk and beef samples, and samples inoculated with the nontarget microorganisms). This method could be used for the rapid detection of E. coli O26 VTEC from foods and for the rapid identification and characterization of clinical and environmental isolates.

scholarly journals A New Ultrasensitive Bioluminescence-Based Method for Assaying Monoacylglycerol Lipase

2021 ◽  
Vol 22 (11) ◽  
pp. 6148
Author(s):  
Matteo Miceli ◽  
Silvana Casati ◽  
Pietro Allevi ◽  
Silvia Berra ◽  
Roberta Ottria ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Kinetic Constants ◽  
New Method ◽  
Monoacylglycerol Lipase ◽  
Enzymatic Cleavage ◽  
Highly Sensitive ◽  
Bioluminescence Assay ◽  
Identification And Characterization ◽  
In Cells

A novel bioluminescent Monoacylglycerol lipase (MAGL) substrate 6-O-arachidonoylluciferin, a D-luciferin derivative, was synthesized, physico-chemically characterized, and used as highly sensitive substrate for MAGL in an assay developed for this purpose. We present here a new method based on the enzymatic cleavage of arachidonic acid with luciferin release using human Monoacylglycerol lipase (hMAGL) followed by its reaction with a chimeric luciferase, PLG2, to produce bioluminescence. Enzymatic cleavage of the new substrate by MAGL was demonstrated, and kinetic constants Km and Vmax were determined. 6-O-arachidonoylluciferin has proved to be a highly sensitive substrate for MAGL. The bioluminescence assay (LOD 90 pM, LOQ 300 pM) is much more sensitive and should suffer fewer biological interferences in cells lysate applications than typical fluorometric methods. The assay was validated for the identification and characterization of MAGL modulators using the well-known MAGL inhibitor JZL184. The use of PLG2 displaying distinct bioluminescence color and kinetics may offer a highly desirable opportunity to extend the range of applications to cell-based assays.

scholarly journals Isolation and characterization of escherichia coli in ready-to-eat foods vended in Islamic University, Kushtia

1970 ◽  
Vol 18 ◽  
pp. 99-103 ◽  
Author(s):  
S Biswas ◽  
MAK Parvez ◽  
M Shafiquzzaman ◽  
S Nahar ◽  
MN Rahman
Keyword(s):  
Escherichia Coli ◽  
Pattern Analysis ◽  
University Campus ◽  
Rflp Pattern ◽  
E Coli ◽  
Fish Meat ◽  
Isolation And Characterization ◽  
Food Borne ◽  
Identification And Characterization

Context: Escherichia coli is shed in the feces of warm blooded animals and humans and thus potential for public health. Detection and characterization of E. coli in the ready-to-eat (RTE) foods concerns due to their presence indicates fecal contamination of the food.   Objective: To identify, characterize and RFLP pattern analysis of E. coli isolated from RTE foods vended in Islamic University campus, Kushtia.   Materials and Methods: Fifty samples from four types of consumed foods in six student halls of residence, some temporary restaurants of Islamic University, Kushtia were assessed for bacterial contamination by standard methods. Identification and characterization of E. coli isolates were performed using IMViC tests. Genomic DNA was used to perform RFLP pattern analysis.   Results: Thirty seven out of 50 (74%) examined samples of RTE foods had E. coli contamination. The highest number of E. coli was isolated from vegetable oriented RTE foods (90.90%) and fish, meat and cereals samples were also significantly E. coli positive. RFLP profiling of two E. coli isolates were observed.   Conclusion: The results of this study provide evidence that some RTE foods had unsatisfactory levels of contamination with E. coli. Thus street vended RTE food could be important potential vehicles for food-borne diseases. Molecular characterization may be exploited to identify food borne pathogen among different species.  Keywords: Ready-to-eat foods; Escherichia coli; RFLP pattern DOI: http://dx.doi.org/10.3329/jbs.v18i0.8783 JBS 2010; 18(0): 99-103

scholarly journals Characterization of a universal screening approach for congenital CMV infection based on a highly-sensitive, quantitative, multiplex real-time PCR assay

PLoS ONE ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. e0227143
Author(s):  
Angela Nagel ◽  
Emmanouela Dimitrakopoulou ◽  
Norbert Teig ◽  
Peter Kern ◽  
Thomas Lücke ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Universal Screening ◽  
Cmv Infection ◽  
Pcr Assay ◽  
Screening Approach ◽  
Highly Sensitive ◽  
Congenital Cmv Infection ◽  
Congenital Cmv

Identification and characterization of four new tra cistrons on the E. coli K12 sex factor F

Plasmid ◽  
1978 ◽  
Vol 1 (3) ◽  
pp. 316-323 ◽  
Author(s):  
T. Miki ◽  
T. Horiuchi ◽  
N.S. Willetts
Keyword(s):  
E Coli ◽  
Identification And Characterization

scholarly journals Postgenomic Scan of Metallo-β-Lactamase Homologues in Rhizobacteria: Identification and Characterization of BJP-1, a Subclass B3 Ortholog from Bradyrhizobium japonicum

2006 ◽  
Vol 50 (6) ◽  
pp. 1973-1981 ◽  
Author(s):  
Magdalena Stoczko ◽  
Jean-Marie Frère ◽  
Gian Maria Rossolini ◽  
Jean-Denis Docquier
Keyword(s):  
Bradyrhizobium Japonicum ◽  
Catalytic Efficiency ◽  
Open Reading Frames ◽  
Human Pathogens ◽  
Microbial Genomes ◽  
E Coli ◽  
Genes Encoding ◽  
And Function ◽  
Identification And Characterization

ABSTRACT The diffusion of metallo-β-lactamases (MBLs) among clinically important human pathogens represents a therapeutic issue of increasing importance. However, the origin of these resistance determinants is largely unknown, although an important number of proteins belonging to the MBL superfamily have been identified in microbial genomes. In this work, we analyzed the distribution and function of genes encoding MBL-like proteins in the class Rhizobiales. Among 12 released complete genomes of members of the class Rhizobiales, a total of 57 open reading frames (ORFs) were found to have the MBL conserved motif and identity scores with MBLs ranging from 8 to 40%. On the basis of the best identity scores with known MBLs, four ORFs were cloned into Escherichia coli for heterologous expression. Among their products, one (blr6230) encoded by the Bradyrhizobium japonicum USDA110 genome, named BJP-1, hydrolyzed β-lactams when expressed in E. coli. BJP-1 enzyme is most closely related to the CAU-1 enzyme from Caulobacter vibrioides (40% amino acid sequence identity), a member of subclass B3 MBLs. A kinetic analysis revealed that BJP-1 efficiently hydrolyzed most β-lactam substrates, except aztreonam, ticarcillin, and temocillin, with the highest catalytic efficiency measured with meropenem. Compared to other MBLs, BJP-1 was less sensitive to inactivation by chelating agents.

scholarly journals Identification and characterization of novel porcine astroviruses (PAstVs) with high prevalence and frequent co-infection of individual pigs with multiple PAstV types

2013 ◽  
Vol 94 (3) ◽  
pp. 570-582 ◽  
Author(s):  
Chao-Ting Xiao ◽  
Luis G. Giménez-Lirola ◽  
Priscilla F. Gerber ◽  
Yong-Hou Jiang ◽  
Patrick G. Halbur ◽  
...  
Keyword(s):  
Zoonotic Potential ◽  
The Novel ◽  
Extraintestinal Manifestations ◽  
Faecal Samples ◽  
High Prevalence ◽  
The Usa ◽  
History Of ◽  
Identification And Characterization ◽  
Sequence Identities

Many astrovirus (AstV) species are associated with enteric disease, although extraintestinal manifestations in mammalian and avian hosts have also been described. In this study, the prevalence rates of porcine AstV types 1–5 (PAstV1–PAstV5) were investigated using faecal samples from 509 pigs of which 488 (95.9 %) came from farms with a history of diarrhoea. All of the five known PAstV types were found to circulate in pigs in the USA, and co-infection of a single pig with two or more PAstV types was frequently observed. A high overall prevalence of 64.0 % (326/509) of PAstV RNA-positive samples was detected, with 97.2 % (317/326) of the PAstV RNA-positive pigs infected with PAstV4. Further genomic sequencing and characterization of the selected isolates revealed low sequence identities (49.2–89.0 %) with known PAstV strains, indicating novel types or genotypes of PAstV2, PAstV4 and PAstV5. Some new features of the genomes of the PAstVs were also discovered. The first complete genome of a PAstV3 isolate was obtained and showed identities of 50.5–55.3 % with mink AstV and the novel human AstVs compared with 38.4–42.7 % with other PAstV types. Phylogenetic analysis revealed that PAstV1, PAstV2 and PAstV3 were more closely related to AstVs from humans and other animals than to each other, indicating past cross-species transmission and the zoonotic potential of these PAstVs.

scholarly journals Identification and Characterization of an Archaeon-Specific Riboflavin Kinase

2008 ◽  
Vol 190 (7) ◽  
pp. 2615-2618 ◽  
Author(s):  
Zahra Mashhadi ◽  
Hong Zhang ◽  
Huimin Xu ◽  
Robert H. White
Keyword(s):  
Escherichia Coli ◽  
Metal Ion ◽  
Biosynthetic Pathway ◽  
Recombinant Expression ◽  
Cell Extract ◽  
Flavin Mononucleotide ◽  
E Coli ◽  
Gene Recombinant ◽  
Identification And Characterization

ABSTRACT The riboflavin kinase in Methanocaldococcus jannaschii has been identified as the product of the MJ0056 gene. Recombinant expression of the MJ0056 gene in Escherichia coli led to a large increase in the amount of flavin mononucleotide (FMN) in the E. coli cell extract. The unexpected features of the purified recombinant enzyme were its use of CTP as the phosphoryl donor and the absence of a requirement for added metal ion to catalyze the formation of FMN. Identification of this riboflavin kinase fills another gap in the archaeal flavin biosynthetic pathway. Some divalent metals were found to be potent inhibitors of the reaction. The enzyme represents a unique CTP-dependent family of kinases.

scholarly journals Identification and Characterization of a New Lipoprotein, NlpI, in Escherichia coli K-12

1999 ◽  
Vol 181 (14) ◽  
pp. 4318-4325 ◽  
Author(s):  
Masaru Ohara ◽  
Henry C. Wu ◽  
Krishnan Sankaran ◽  
Paul D. Rick
Keyword(s):  
Escherichia Coli ◽  
Direct Consequence ◽  
Insertion Mutant ◽  
Polynucleotide Phosphorylase ◽  
Wild Type ◽  
E Coli ◽  
K 12 ◽  
Identification And Characterization ◽  
Lines Of Evidence

ABSTRACT We report here the identification of a new lipoprotein, NlpI, inEscherichia coli K-12. The NlpI structural gene (nlpI) is located between the genes pnp(polynucleotide phosphorylase) and deaD (RNA helicase) at 71 min on the E. coli chromosome. The nlpI gene encodes a putative polypeptide of approximately 34 kDa, and multiple lines of evidence clearly demonstrate that NlpI is indeed a lipoprotein. An nlpI::cm mutation rendered growth of the cells osmotically sensitive, and incubation of the insertion mutant at an elevated temperature resulted in the formation of filaments. The altered phenotype of the mutant was a direct consequence of the mutation in nlpI, since it was complemented by the wild-type nlpI gene alone. Overexpression of the unaltered nlpI gene in wild-type cells resulted in the loss of the rod morphology and the formation of single prolate ellipsoids and pairs of prolate ellipsoids joined by partial constrictions. NlpI may be important for an as-yet-undefined step in the overall process of cell division.

scholarly journals ISOLATION AND CHARACTERIZATION OF ESCHERICHIA COLI FROM BUFFALO CALVES IN SOME SELECTED AREAS OF BANGLADESH

1970 ◽  
Vol 8 (1) ◽  
pp. 23-26 ◽  
Author(s):  
SK Paul ◽  
MSR Khan ◽  
MA Rashid ◽  
J Hassan ◽  
SMS Mahmud
Keyword(s):  
Escherichia Coli ◽  
Nalidixic Acid ◽  
Rectal Swab ◽  
Antibiotic Sensitivity ◽  
Buffalo Calves ◽  
E Coli ◽  
Isolation And Characterization ◽  
Sensitivity Pattern ◽  
Highly Sensitive

The research works was conducted with a view to isolate and identify the Escherichia coli (E. coli) organism from diarrhoeic cases of buffalo reared in selected areas of Bangladesh as well the prevalence and antibiotic sensitivity pattern of the isolated E. coli in the Department of Microbiology and Hygiene, Bangladesh Agricultural University (BAU), Mymensingh-2202 during the period from April 2008 to May 2009. A total of 50 rectal swab samples were collected from 4 different places namely Haluaghat and Boira of Mymensingh, Madupur of Tangail and Kazipur of Sirajgonj districts. The samples were aseptically carried to the laboratory of the Department of Microbiology and Hygiene and subjected to different cultural, morphological and biochemical examinations. Upon cultural, morphological and biochemical examinations 23 (45%) samples were found to be positive for E. coli. The highest prevalence was found in Haluaghat, Mymensingh (53.33%) and the lowest (40.00%) in Boira, Mymensingh and Kazipur, Sirajganj. Antibiogram study revealed that the isolated E. coli was highly sensitive to Enrofloxacin and Ciprofloxacin, moderately sensitive to Cefalexin and Amoxicillin, and resistant to Nalidixic acid and Erythromycin. DOI = 10.3329/bjvm.v8i1.7398 Bangl. J. Vet. Med. (2010). 8(1): 23-26

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