scholarly journals AVIAN INFLUENZA ANTIBODY RESPONSE IN PIGLETS THATAREGIVEN ESCHERICHIA COLI–AVIAN INFLUENZA VACCINES

2021 ◽  
Vol 10 (1) ◽  
pp. 61-70
Author(s):  
Audrey Febiannya Putri Bhaskara ◽  
I Gusti Ngurah Kade Mahardika ◽  
I Nyoman Suartha
Keyword(s):  
Escherichia Coli ◽  
Avian Influenza ◽  
Optical Density ◽  
Antibody Test ◽  
Influenza Vaccines ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Antibody ◽  
E Coli ◽  
Test Kit ◽  
Virus Influenza

Babi berperan penting dalam ekologi virus influenza, karena babi dapat berperan sebagai wahana untuk reasorsi virus influenza dari unggas dan mamalia. Vaksinasi dengan antigen influenza universal, yaitu nukleoprotein, dapat menurunkan peluang babi dalam memunculkan virus influenza baru. Penelitian ini bertujuan untuk mengentahui respons antibodi dari vaksinasi dengan nukleoprotein rekombinan - Escherichia coli. Sebanyak 12 anak babi landrace dari tiga induk yang berbeda dipilih secara acak. Enam ekor divaksinasi dengan vaksin nucleoprotein-E. coli pada umur tujuh hari dan diulang pada umur 21 hari. Enam ekor tidak divaksinasi. Serum diambil pada umur 35 hari. Nilai optical density (OD) antibodi terhadap nukleoprotein diuji dengan teknik Enzyme-Linked Immunosorbent Assay (ELISA) dengan menggunakan Kit ELISA komersial, Avian Influenza Virus Antibody Test Kit. Hasil penelitian menunjukkan bahwa nilai Optical Density rata-rata babi yang divaksinasi (0,367) secara statistika nyata lebih tinggi dibandingkan dengan yang tidak divaksinasi (0,054). Vaksin rekombinan nucleoprotein-E. coli yang dicobakan mampu meningkatan antibodi terhadap virus avian influenza pada anak babi.

scholarly journals Turntable Paper-Based Device to Detect Escherichia coli

Micromachines ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 194
Author(s):  
Yung-Chih Wang ◽  
Yao-Hung Tsai ◽  
Ching-Fen Shen ◽  
Ming-Yao He ◽  
Yi-Chen Fu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Infectious Diseases ◽  
Point Of Care ◽  
Multiple Integral ◽  
Enzyme Linked Immunosorbent Assay ◽  
Considerable Time ◽  
E Coli ◽  
Colony Forming Unit ◽  
Experimental Implementation ◽  
Colony Forming Units

Escherichia coli has been known to cause a variety of infectious diseases. The conventional enzyme-linked immunosorbent assay (ELISA) is a well-known method widely used to diagnose a variety of infectious diseases. This method is expensive and requires considerable time and effort to conduct and complete multiple integral steps. We previously proposed the use of paper-based ELISA to rapidly detect the presence of E. coli. This approach has demonstrated utility for point-of-care (POC) urinary tract infection diagnoses. Paper-based ELISA, while advantageous, still requires the execution of several procedural steps. Here, we discuss the design and experimental implementation of a turntable paper-based device to simplify the paper-based ELISA protocols for the detection of E. coli. In this process, antibodies or reagents are preloaded onto zones of a paper-based device and allowed to dry before use. We successfully used this device to detect E. coli with a detection limit of 105 colony-forming units (colony-forming unit [CFU])/mL.

An Improved Screening Method for the Detection and Isolation of Escherichia coli 0157:H7 From Meat, Incorporating the 3M Petrifilm™ Test Kit - HEC - for Hemorrhagic Escherichia coli 0157:H7

1990 ◽  
Vol 53 (11) ◽  
pp. 936-940 ◽  
Author(s):  
ANITA J. G. OKREND ◽  
BONNIE E. ROSE ◽  
RICHARD MATNER
Keyword(s):  
Escherichia Coli ◽  
Screening Program ◽  
Enrichment Culture ◽  
Screening Method ◽  
Meat Sample ◽  
E Coli ◽  
Enrichment Cultures ◽  
Poultry Products ◽  
Test Kit ◽  
Cooked Meat

A screening method was devised incorporating a commercially available reactive disc blot ELISA for Escherichia coli 0157 antigen, into a cultural screening program for the isolation of E. coli 0157:H7 from meat and poultry products. The method includes the inoculation of a raw or cooked meat sample into an enrichment broth, incubation with shaking at 37°C for 6 to 8 h, followed by inoculation of 3M Petrifilm™ E. coli Count plates with dilutions of the enrichment culture. The Petrifilm plates were incubated at 42°C for 18 h and tested for the presence of the 0157 antigen. The enrichment cultures were reincubated static at 35°C after the initial shaken incubation. Isolation was attempted from the positive Petrifilm plates by both a direct picking and streaking method and by the 3M Prompt™ isolation method. Isolation also was attempted from the 24-h enrichment cultures by spread plating serial dilutions on 150 × 15 mm MacConkey sorbitol agar (MSA) and MSA with 5-bromo-4-chloro-3-indoxyl-β-D-glucuronic acid cyclohexylammonium salt (BCIG). This fast and efficient screening procedure identifies negative and presumptive positive samples in 26–28 h. Isolation and confirmation of the presumptive positive isolates require an additional 3 to 4 d.

scholarly journals A PCR-Based Method of Detection and Differentiation of K88+ Adhesive Escherichia Coli

1996 ◽  
Vol 8 (4) ◽  
pp. 460-463 ◽  
Author(s):  
Mark A. Franklin ◽  
David H. Francis ◽  
Diane Baker ◽  
Alan G. Mathew
Keyword(s):  
Escherichia Coli ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigenic Variant ◽  
Variable Regions ◽  
E Coli ◽  
Clinical Assay ◽  
Pcr Product ◽  
Structural Subunit ◽  
Pcr Products ◽  
Polymerase Chain

The objective of this study was to develop a polymerase chain reaction (PCR)-based method to detect and differentiate among Escherichia coli strains containing genes for the expression of 3 antigenic variants of the fimbrial adhesin K88 (K88ab, K88ac, and K88ad). Five primers were designed that allowed detection of K88+ E. coli, regardless of antigenic variant, and the separate detection of the ab, ac, and ad variants. Primers AM005 and AM006 are 21 base pair (bp) oligomers that correspond to a region of the K88 operon that is common to all 3 antigenic variants. Primers MF007, MF008, and MF009 are 24-bp oligomers that matched variable regions specific to ab, ac, and ad, respectively. Using primers AM005 and AM006, a PCR product was obtained that corresponds to a 764-bp region within the large structural subunit of the K88 operon common to all 3 antigenic variants. Primer AM005 used with MF007, MF008, or MF009 produced PCR products approximately 500-bp in length from within the large structural subunit of the K88 operon of the 3 respective antigenic variants. Fragments were identified by rates of migration on a 1% agarose gel relative to each other as well as to BstEII-digested lambda fragments. This PCR-based method was comparable to the enzyme-linked immunosorbent assay and western blot test in the ability to differentiate between the antigenic variants. K88+ E. coli were differentiated from among laboratory strains and detected in ileal samples taken from cannulated pigs challenged with a known K88+ variant. K88+ E. coli were also detected from fecal swabs taken from newly weaned pigs, thus confirming that this PCR-based test could provide a convenient clinical assay for the detection of K88+ E. coli. Detection and differentiation of K88+ E. coli using general and specific primers was successful. PCR methods of detection should permit identification of K88+ antigenic variants regardless of the level of expression of the antigen.

scholarly journals The combination of Elephantopus scaber and Sauropus androgynus promotes erythroid lineages and modulates follicle-stimulating hormone and luteinizing hormone levels in pregnant mice infected with Escherichia coli

2021 ◽  
pp. 1398-1404
Author(s):  
Muhammad Sasmito Djati ◽  
Yuyun Ika Christina ◽  
Muhaimin Rifa'i
Keyword(s):  
Escherichia Coli ◽  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Relative Number ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hormone Levels ◽  
E Coli ◽  
Pregnant Mice ◽  
Elephantopus Scaber ◽  
Stimulating Hormone

Background and Aim: Escherichia coli infection produces an adverse effect on the erythrocyte lineage and hormone levels during pregnancy. This study aimed to evaluate the effects of Elephantopus scaber (ES) and Sauropus androgynus (SA) in combination on circulating follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels and erythropoiesis changes in E. coli-infected pregnant mice. Materials and Methods: Female Balb/c mice were mated with normal male mice and pregnancies were identified by the formation of vaginal plugs. Twenty-eight pregnant mice were divided randomly into seven groups: A control group (N), E. coli-infected pregnant mice (K+), and infected pregnant mice received the following five treatments: (1) Only ES; (2) ESSA1 (75:25); (3) ESSA2 (50:50); (4) ESSA3 (25:75); and (5) only SA, beginning from the 1st to the 16th day of pregnancy. Pregnant mice were infected with 107 CFU/mL of E. coli on day 4. Blood serum was collected on days 8, 12, and 16 of pregnancy and LH and FSH levels were measured by enzyme-linked immunosorbent assay. Bone marrow was isolated to determine the relative number of TER-119+VLA4+ and TER-119+CD34+ using flow cytometry. Results: The ESSA1 and SA groups exhibited a marked increase in LH levels. The combination of ES and SA administered at a 25:75 ratio (ESSA3) altered FSH levels and the relative number of TER-119+VLA4+ in infected pregnant mice. Combined with SA at an equal ratio (50:50), ESSA2 group exhibited a significant increase in the expression of TER119+CD34+ compared with the other treatment groups. Conclusion: ES and SA combined at a ratio of 25:75 exhibited optimal results in altering hormonal and erythropoiesis in infected pregnant mice.

scholarly journals Detection of avian reovirus antibodies in layer birds of small scale commercial farms in Dinajpur district of Bangladesh

2016 ◽  
Vol 1 (3) ◽  
pp. 612-621 ◽  
Author(s):  
Abdus Salam ◽  
Md Atiqul Haque ◽  
Md Mostafizer Rahman ◽  
Mir Rowshan Akter ◽  
Farzana Afroz
Keyword(s):  
Antibody Titer ◽  
Specific Antibody ◽  
Age Groups ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Scale ◽  
Avian Reovirus ◽  
Serum Samples ◽  
Aged Group ◽  
Test Kit

The present study was conducted on layer birds of different age groups to determine specific antibody titer level against avian reovirus (ARV) by indirect enzyme linked immunosorbent assay (iELISA) at Dinajpur district of Bangladesh. This study showed that ARV specific antibody positive cases were 84 out of 90 blood serum samples and the highest antibody titer was 26120 and lowest antibody titer was 288. The total 93.33% sera samples were showed positive result. The study showed that 100% sera sample were positive against ARV at 6 weeks of aged group and the highest, lowest and mean antibody titer were 13917, 4895 and 10269 respectively. On the other hand 88.88% sera sample were positive against ARV at 10 weeks of aged group and the highest, lowest and mean antibody titer were 9779, 288 and 5689.89 respectively. The sera sample collected from 14 weeks of aged group showed 88.88% positive and the highest, lowest and mean antibody titer were 11727, 871 and 5250 respectively. The sera sample collected from 18 weeks of aged group showed 88.88% positive against ARV and the highest, lowest and mean antibody titer were 24440, 1234 and 12648.89 respectively. The sera sample collected from 22 weeks of aged group were 100% positive against ARV and the highest, lowest and mean antibody titer were 26120, 1752 and 11373.89 respectively. The sera sample collected from 26 weeks of aged group showed 100% positive against ARV and the highest, lowest and mean antibody titer were 8566, 1630 and 4327.44 respectively. The sera sample collected from 30 weeks of aged group showed 100% positive against ARV and the highest, lowest and mean antibody titer were 13431, 1989 and 5890.56 respectively. The sera sample collected from 40 weeks of aged group showed 77.77% positive against ARV and the highest, lowest and mean antibody titer were 14618, 433 and 5103.22 respectively. The sera sample collected from 48 weeks of aged group showed 88.88% positive against ARV and the highest, lowest and mean antibody titer were 14553, 957 and 7436.5 respectively. In conclusion it is evident that avian reovirus-specific antibody was successfully detected through commercially available avian reovirus antibody test kit (ELISA kit) and the virus induced a significant antibody titer indicating the affecting virus was absolutely ARV.Asian J. Med. Biol. Res. December 2015, 1(3): 612-621

Comparison of Rapid Enzyme-Linked Immunosorbent Assay and Immunomagnetic Separation Methods for Detection of Escherichia coli O157 in Fecal, Hide, Carcass, and Ground Beef Samples

2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Immunomagnetic Separation ◽  
Separation Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Escherichia Coli O157 ◽  
Perfect Agreement ◽  
Fecal Samples ◽  
E Coli ◽  
Beef Products

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.

Use of a Shiga Toxin (Stx)-Enzyme-Linked Immunosorbent Assay and Immunoblot for Detection and Isolation of Stx-Producing Escherichia coli from Naturally Contaminated Beef

2000 ◽  
Vol 63 (9) ◽  
pp. 1167-1172 ◽  
Author(s):  
HEBA NASHED ATALLA ◽  
ROGER JOHNSON ◽  
SCOTT MCEWEN ◽  
R. W. USBORNE ◽  
C. L. GYLES
Keyword(s):  
Escherichia Coli ◽  
Vero Cell ◽  
Immunosorbent Assay ◽  
Shiga Toxin ◽  
Total Coliform ◽  
Complete Agreement ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spearman Correlation ◽  
Southern Ontario ◽  
E Coli

The purpose of this study was to evaluate an enzyme-linked immunosorbent assay (ELISA) and an immunoblot procedure for detection and isolation of Shiga toxin-producing Escherichia coli (STEC) from beef, and to correlate the presence of STEC in beef with E. coli and total coliform counts. A total of 120 samples of boneless beef supplied to a meat processor in southern Ontario were tested for the presence of STEC, E. coli, and total coliforms. Following enrichment in modified tryptic soy broth, samples were screened for Shiga toxin (Stx) by a Stx-ELISA and a Vero cell assay (VCA). Samples that were positive in the Stx-ELISA were subjected to the Stx-immunoblot for STEC isolation. Overall, 33.3% of samples were positive in the VCA, and 34.2% were positive in the Stx-ELISA. There was almost complete agreement between the Stx-ELISA and the VCA results (kappa = 0.98). The sensitivity and specificity of the Stx-ELISA with respect to the VCA were 100% and 98.75%, respectively. STEC were isolated by the Stx-immunoblot from 87.8% of the samples that were positive in the Stx-ELISA. The STEC isolates belonged to 19 serotypes, with serotype O113:H21 accounting for 10 of 41 isolates. No STEC of serotype O157:H7 were isolated. There was a significant correlation between E. coli counts and total coliform counts (Spearman correlation coefficient = 0.68, P < 0.01). The E. coli count was positively correlated with detection of STEC by both the Stx-ELISA and the VCA (P < 0.01).

Evaluation of Three Commercially Available Enzyme-Linked Immunosorbent Assay Kits for Detection of Shiga Toxin

2009 ◽  
Vol 72 (4) ◽  
pp. 741-747 ◽  
Author(s):  
JOHN WILLFORD ◽  
KENNETH MILLS ◽  
LAWRENCE D. GOODRIDGE
Keyword(s):  
Escherichia Coli ◽  
Immunosorbent Assay ◽  
Shiga Toxin ◽  
Screening Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microplate Assay ◽  
E Coli ◽  
Elisa Kit ◽  
Pure Cultures ◽  
Order Of Magnitude

Three commercially available Shiga toxin (Stx) enzyme-linked immunosorbent assay (ELISA) kits were evaluated for their ability to detect Stx in pure cultures of Stx-producing Escherichia coli (specificity). The detection limits (sensitivity) of each ELISA kit were also evaluated. Seventy-eight Stx-producing E. coli (STEC) isolates that produced Stx1, Stx2, or Stx1 and Stx2 variants were examined in this study. The specificities of the tests were comparable, and the sensitivities of two of the tests (Premier EHEC and rBiopharm Ridascreen Verotoxin Enzyme Immunoassay) were within the same order of magnitude. The ProSpecT Shiga Toxin E. coli Microplate Assay was approximately 10-fold less sensitive. The inability of all three tests to detect the Stx2d and Stx2e variants indicated that some STEC strains may not be detected by Stx ELISA. The ability of the Premier EHEC ELISA to detect toxin in artificially inoculated bovine fecal samples (following enrichment) indicated that this kit may be used to screen cattle for the presence of Stx as an indicator of the presence of STEC. In particular, such a screening method could be useful during the summer, when the number of STEC-positive animals and the number of STEC that they shed increase.

scholarly journals THE EFFECTS OF SULFATHIAZOLE AND OF PROPYL CARBAMATE ON THE RATE OF OXYGEN CONSUMPTION AND GROWTH IN ESCHERICHIA COLI

1947 ◽  
Vol 30 (3) ◽  
pp. 263-278 ◽  
Author(s):  
Kenneth C. Fisher ◽  
Florence H. Armstrong
Keyword(s):  
Escherichia Coli ◽  
Oxygen Consumption ◽  
Cell Count ◽  
Optical Density ◽  
Viable Cell ◽  
E Coli ◽  
Rate Of Growth ◽  
A Value ◽  
Rate Of Oxygen Consumption ◽  
Rates Of Growth

1. The rates of growth and of oxygen consumption by cells of E. coli have been measured under identical conditions, and the effects of sulfathiazole (ST) and of n-propyl carbamate (PC) on these two processes have been compared. 2. The rate of growth was measured by (a) the increase in the viable cell count, (b) the increase in the optical density of the culture, (c) the increase in the rate of oxygen consumption, and (d) the decrease in the ammonia of the medium. The results as indicated by these several measures were identical under the conditions of these experiments. 3. Concentrations of ST or of PC which are just sufficient to stop growth completely, lower the rate of oxygen consumption per unit of bacterial protoplasm to a value approximately 50 per cent of that seen in the absence of the inhibitor. 4. It is shown that the rate of oxygen consumption in cells from old cultures is less affected by ST than is the rate of oxygen consumption by cells from young cultures. It is probable that the rate of oxygen consumption by "old" cells is lower than that of "young" cells. 5. The effects of ST and PC on both the rate of oxygen consumption and the rate of growth are very similar, indicating in a general way, that the mechanism of the actions of these two inhibitors is similar. Furthermore, since both of them produce appreciable inhibition of the rate of oxygen consumption while they are inhibiting growth, the possibility that the effect on oxygen consumption is the immediate cause of the effect on growth must be entertained.

REGULATION OF ACETOHYDROXY ACID SYNTHETASE IN STREPTOMYCIN-DEPENDENT ESCHERICHIA COLI K-12

1966 ◽  
Vol 44 (5) ◽  
pp. 607-612 ◽  
Author(s):  
W. J. Polglase
Keyword(s):  
Escherichia Coli ◽  
Optical Density ◽  
Density Increase ◽  
Regulatory Effect ◽  
Michaelis Constant ◽  
E Coli ◽  
K 12 ◽  
Positive Effect

The formation of acetohydroxy acid (AHA) synthetase in streptomycin-dependent Escherichia coli K-12 as a function of antibiotic (dihydrostreptomycin) concentration can be expressed by a Michaelis constant (KM = 2 × 10−4M). Valine exerts end-product repression (partial) of AHA synthetase in competition with the positive regulatory effect of dihydrostreptomycin. In dependent E. coli K-12, growth (as measured by optical density increase) correlated closely with the formation of AHA synthetase. The observed positive effect of dihydrostreptomycin on AHA synthetase formation suggests that derepression of this enzyme may be obligatory in antibiotic-dependent cells.

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