scholarly journals Microbiota of Sardinian Goat’s Milk and Preliminary Characterization of Prevalent LAB Species for Starter or Adjunct Cultures Development

2019 ◽  
Vol 2019 ◽  
pp. 1-7 ◽  
Author(s):  
Maria Barbara Pisano ◽  
Maura Deplano ◽  
Maria Elisabetta Fadda ◽  
Sofia Cosentino
Keyword(s):  
Starter Cultures ◽  
Technological Properties ◽  
E Coli ◽  
Adjunct Cultures ◽  
Goat’S Milk ◽  
Good Potential ◽  
Goat's Milk ◽  
Coagulase Positive Staphylococci

This work was performed to study the microbiota of raw goat’s milk (67 samples) collected in different areas of Sardinia, in order to select autochthonous lactic acid bacteria (LAB) strains for use in goat cheese manufacturing. Total mesophilic bacteria ranged between 105 and 107 cfu/mL; mean counts of Enterobacteriaceae did not exceed 4 log cfu/mL whereas those of E. coli and coagulase-positive staphylococci were lower than 1.5 and 2 log ufc /ml, respectively. Neither Salmonella spp. nor Listeria monocytogenes were recovered. The numbers of total LAB were in the range from 104 to 107 cfu/mL and mean yeasts counts varied between 103 and 105 cfu/mL. The most frequently isolated LAB species were Lactococcus lactis subsp. lactis and Lactobacillus paracasei. The presence of Enterococcus faecium was also noteworthy. The in vitro study of some functional characteristics related to technological properties of the strains belonging to these species allowed to point out some strains possessing good potential for use as adjunct or starter cultures in the production of cheese.

Microbiological Characterization of Randomly Selected Portuguese Raw Milk Cheeses with Reference to Food Safety

2007 ◽  
Vol 70 (7) ◽  
pp. 1710-1716 ◽  
Author(s):  
GONÇALO ALMEIDA ◽  
ALEXANDRE FIGUEIREDO ◽  
MARTA RÔLA ◽  
RUI MANUEL BARROS ◽  
PAUL GIBBS ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Raw Milk ◽  
Processing Conditions ◽  
Indicator Organisms ◽  
Soft Or ◽  
E Coli ◽  
Goat’S Milk ◽  
Goat's Milk ◽  
Made In

Seventy raw milk cheeses made in different regions of Portugal, both hard and soft varieties, made with cow's, ewe's, or goat's milk or combinations of these, were sampled within their quoted shelf lives for microbiological safety. On the basis of the presence or numbers of Escherichia coli, E. coli O157, Staphylococcus aureus, Salmonella, and Listeria monocytogenes, cheeses were categorized as satisfactory, acceptable, unsatisfactory, or unacceptable and potentially hazardous. Twenty-two of the 70 cheeses were classified as satisfactory or acceptable. Thirty-seven of the cheeses were considered unsatisfactory because of the presence of E. coli, S. aureus, or both, while 11 of the cheeses were graded as unacceptable and potentially hazardous because of the presence of excessive numbers of S. aureus, E. coli, or L. monocytogenes and the presence of Salmonella in three of these. All cheeses graded as unacceptable and potentially hazardous were soft or semisoft cheeses made with ewe's and goat's milk, with the exception of two hard cheeses made with cow's milk. E. coli O157 was not detected in any of the cheeses. According to the present results, it seems that the presence or counts of pathogenic or indicator organisms in raw milk cheeses cannot be related to the processing conditions, milk type, or region of production.

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

2019 ◽  
Author(s):  
Priya Prakash ◽  
Travis Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Amyloid Beta ◽  
Western Blots ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
Set Up ◽  
Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

scholarly journals Functional identification of ygiP as a positive regulator of the ttdA-ttdB-ygjE operon

Microbiology ◽  
2006 ◽  
Vol 152 (7) ◽  
pp. 2129-2135 ◽  
Author(s):  
Taku Oshima ◽  
Francis Biville
Keyword(s):  
Functional Characterization ◽  
Anaerobic Growth ◽  
Functional Identification ◽  
New Members ◽  
E Coli ◽  
Inner Membrane Protein ◽  
Tartrate Dehydratase

Functional characterization of unknown genes is currently a major task in biology. The search for gene function involves a combination of various in silico, in vitro and in vivo approaches. Available knowledge from the study of more than 21 LysR-type regulators in Escherichia coli has facilitated the classification of new members of the family. From sequence similarities and its location on the E. coli chromosome, it is suggested that ygiP encodes a lysR regulator controlling the expression of a neighbouring operon; this operon encodes the two subunits of tartrate dehydratase (TtdA, TtdB) and YgiE, an integral inner-membrane protein possibly involved in tartrate uptake. Expression of tartrate dehydratase, which converts tartrate to oxaloacetate, is required for anaerobic growth on glycerol as carbon source in the presence of tartrate. Here, it has been demonstrated that disruption of ygiP, ttdA or ygjE abolishes tartrate-dependent anaerobic growth on glycerol. It has also been shown that tartrate-dependent induction of the ttdA-ttdB-ygjE operon requires a functional YgiP.

Characterization of in vitro and in vivo murine models of human enteropathogenic E. coli (EPEC) infection

2003 ◽  
Vol 124 (4) ◽  
pp. A558
Author(s):  
Suzana D. Savkovic ◽  
Farol L. Tomson ◽  
Michelle Muza ◽  
Gail Hecht
Keyword(s):  
Murine Models ◽  
E Coli

scholarly journals Characterization of MazFSa, an Endoribonuclease from Staphylococcus aureus

2007 ◽  
Vol 189 (24) ◽  
pp. 8871-8879 ◽  
Author(s):  
Zhibiao Fu ◽  
Niles P. Donegan ◽  
Guido Memmi ◽  
Ambrose L. Cheung
Keyword(s):  
Staphylococcus Aureus ◽  
Environmental Stress Response ◽  
Binding Studies ◽  
Cell Growth Arrest ◽  
Consensus Sequences ◽  
E Coli ◽  
Endoribonuclease Activity

ABSTRACT The mazEF homologs of Staphylococcus aureus, designated mazEFsa , have been shown to cotranscribe with the sigB operon under stress conditions. In this study, we showed that MazEF Sa , as with their Escherichia coli counterparts, compose a toxin-antitoxin module wherein MazF Sa leads to rapid cell growth arrest and loss in viable CFU upon overexpression. MazF Sa is a novel sequence-specific endoribonuclease which cleaves mRNA to inhibit protein synthesis. Using ctpA mRNA as the model substrate both in vitro and in vivo, we demonstrated that MazF Sa cleaves single-strand RNA preferentially at the 5′ side of the first U or 3′ side of the second U residue within the consensus sequences VUUV′ (where V and V′ are A, C, or G and may or may not be identical). Binding studies confirmed that the antitoxin MazE Sa binds MazF Sa to form a complex to inhibit the endoribonuclease activity of MazF Sa . Contrary to the system in E. coli, exposure to selected antibiotics augmented mazEFsa transcription, akin to what one would anticipate from the environmental stress response of the sigB system. These data indicate that the mazEF system of S. aureus differs from the gram-negative counterparts with respect to mRNA cleavage specificity and antibiotic stresses.

Effect In Vitro of Antiparasitic Drugs on Microbial Inhibitor Test Responses for Screening Antibiotic Residues in Goat's Milk

2015 ◽  
Vol 78 (9) ◽  
pp. 1756-1759 ◽  
Author(s):  
T. ROMERO ◽  
M. C. BELTRÁN ◽  
W. REYBROECK ◽  
M. P. MOLINA
Keyword(s):  
Antibiotic Residues ◽  
Goat’S Milk ◽  
Drug Concentrations ◽  
Test Microorganism ◽  
Antiparasitic Drug ◽  
Goat's Milk ◽  
Antiparasitic Drugs ◽  
Almost All ◽  
Positive Results

Microbial inhibitor tests are widely used to screen antibiotic residues in milk; however, these tests are nonspecific and may be affected by various substances capable of inhibiting the growth of the test microorganism. The objective of this study was to determine the effect of antiparasitic drugs in goat's milk on the microbial inhibitor test response. Raw antibiotic-free milk from Murciano-Granadina goats was supplemented with eight concentrations of seven antiparasitic substances (albendazole, 10 to 170 mg/kg; closantel, 1 to 140 mg/kg; diclazuril, 8 to 45 mg/kg; febendazole, 10 to 140 mg/kg; levamisole, 40 to 440 mg/kg; diazinon, 8 to 45 mg/kg; and ivermectin, 40 to 200 mg/kg). Twelve replicates for each concentration were analyzed with three microbial inhibitor tests: BRT MRL, Delvotest SP-NT MSC, and Eclipse 100. The results were interpreted visually (negative or positive). Using a logistic regression model, the concentrations of the antiparasitic drugs producing 5% (IC5), 10% (IC10), and 50% (IC50) positive results were determined. In general, the Eclipse 100 test was less sensitive to the effect of antiparasitic substances; the inhibitory concentrations of almost all the drugs assayed were higher than those for other tests. Conversely, the BRT MRL test was most affected, with high levels of interference at lower antiparasitic drug concentrations. Closantel and diazinon interfered with all microbial tests at lower concentrations than did other drugs (IC5 = 1 to 26 and 12 to 20 mg/kg, respectively), and higher concentrations of levamisole and diclazuril (IC5 = 30 to 240 and 50 to 117 mg/kg, respectively) were required to produce 5% positive results. These findings indicate that microbial inhibitor tests can be affected by elevated concentrations of antiparasitic drugs in goat's milk.

Characterization of a reuterin-producing Lactobacillus coryniformis strain isolated from a goat's milk cheese

2005 ◽  
Vol 104 (3) ◽  
pp. 267-277 ◽  
Author(s):  
R. Martín ◽  
M. Olivares ◽  
M.L. Marín ◽  
J. Xaus ◽  
L. Fernández ◽  
...  
Keyword(s):  
Goat’S Milk ◽  
Goat's Milk ◽  
Lactobacillus Coryniformis

scholarly journals Characterization of the DNA- and Metal-Binding Properties of Vibrio anguillarum Fur Reveals Conservation of a Structural Zn2+ Ion

2000 ◽  
Vol 182 (21) ◽  
pp. 6264-6267 ◽  
Author(s):  
Ekaterina E. Zheleznova ◽  
Jorge H. Crosa ◽  
Richard G. Brennan
Keyword(s):  
Metal Binding ◽  
Iron Uptake ◽  
Vibrio Anguillarum ◽  
Zinc Ion ◽  
Ferric Uptake Regulator ◽  
Binding Properties ◽  
E Coli ◽  
Solution Studies

ABSTRACT The ferric uptake regulator, Fur, represses iron uptake and siderophore biosynthetic genes under iron-replete conditions. Here we report in vitro solution studies on Vibrio anguillarum Fur binding to the consensus 19-bp Escherichia coli iron box in the presence of several divalent metals. We found that V. anguillarum Fur binds the iron box in the presence of Mn2+, Co2+, Cd2+, and to a lesser extent Ni2+ but, unlike E. coli Fur, not in the presence of Zn2+. We also found that V. anguillarum Fur contains a structural zinc ion that is necessary yet alone is insufficient for DNA binding.

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