scholarly journals PCR-based detection of cow’s milk in goat and sheep cheeses marketed in the Czech Republic

2011 ◽  
Vol 24 (No. 3) ◽  
pp. 127-132 ◽  
Author(s):  
E. Mašková ◽  
I. Paulíčková
Keyword(s):  
Mitochondrial Gene ◽  
Retail Trade ◽  
Animal Origin ◽  
The Czech Republic ◽  
Goat Cheese ◽  
Gene Coding ◽  
Reverse Primer ◽  
Pcr Method ◽  
Italian Origin ◽  
Species Specific

A method based on the polymerase chain reaction (PCR) principle was validated for detecting cow’s milk in goat and sheep cheeses. DNA was isolated from the cheeses using the isolation kit Invisorb Spin Food I by Invitek Co., designed for the samples of animal origin. The PCR method applied utilizes the sequence of the mitochondrial gene coding cytochrome b which is specific for mammals. It uses the common forward primer and the reverse primer species-specific. After electrophoresis, cow DNA was characterised by the fragment of the size of 274 bp, goat DNA by the fragment of 157 bp, and sheep DNA by the fragment of 331 bp. The detection limit of the PCR method described (1%) was determined with model samples made from pure goat cheese with a defined addition of cheese made from cow’s milk. The method validated was applied in the analysis of 17 goat cheeses and 7 sheep cheeses obtained from retail trade. Products of Czech, Slovak, French, Dutch, and Italian origin were examined. The presence of undeclared cow’s milk was detected in three kinds of goat cheese and in one of sheep cheese.  

Improvements to a PCR-Based Serogrouping Scheme for Salmonella enterica from Dairy Farm Samples

2015 ◽  
Vol 78 (6) ◽  
pp. 1182-1185 ◽  
Author(s):  
JEFFREY S. KARNS ◽  
BRADD J. HALEY ◽  
JO ANN S. VAN KESSEL
Keyword(s):  
Salmonella Enterica ◽  
Dairy Farm ◽  
Disease Outbreaks ◽  
In Silico Analysis ◽  
Pcr Assay ◽  
Molecular Serotyping ◽  
Reverse Primer ◽  
Pcr Method ◽  
Silico Analysis

Molecular serotyping through the use of PCR is a simple and useful technique for characterizing isolates of Salmonella enterica subsp. enterica belonging to serogroups B, C1, C2, D1, and E1, which are the majority of the isolates associated with human disease outbreaks. However, many of the Salmonella strains currently isolated from dairy farms in the northeastern United States are serovar Cerro, a group K strain not detected by this assay. Primers from a well-known PCR assay for the identification of Salmonella were added to a commonly used serotyping assay so that strains, such as Salmonella Cerro, that do not produce bands in the original assay can be confirmed as belonging to S. enterica subsp. enterica. The modified assay frequently misidentified the serogroup of Salmonella Mbandaka isolates because of failure to amplify the wzxC1 amplicon. Therefore, the reverse primer for the wzxC1 target was modified based on in silico analysis to provide consistent classification of Salmonella Mbandaka as belonging to serogroup C1. These two modifications to the serogrouping PCR method enhance the utility of the method for characterizing Salmonella isolates.

Species delimitation in the leaf beetle genus Macroplea (Coleoptera, Chrysomelidae) based on mitochondrial DNA, and phylogeographic considerations

2006 ◽  
Vol 37 (4) ◽  
pp. 467-479 ◽  
Author(s):  
Gregor Kölsch ◽  
Bo Vest Pedersen ◽  
Olof Biström
Keyword(s):  
Phylogenetic Analysis ◽  
Mitochondrial Dna ◽  
Species Delimitation ◽  
Mitochondrial Gene ◽  
Leaf Beetle ◽  
Genetic Distances ◽  
Amino Acid Substitutions ◽  
Species Status ◽  
Leaf Beetles ◽  
Gene Coding

AbstractThe genus Macroplea Samouelle, 1819 is a group of highly specialized aquatic leaf beetles occurring in the Palaearctic. Since the members of this genus are morphologically very similar, we addressed the question of species identification and delimitation by analysing the second half of the mitochondrial gene coding for the cytochrome oxidase I (COI) subunit. Species limits are inferred from the multimodal frequency distribution of genetic distances between specimens: low genetic distances within a species are clearly set apart from distances between species. The species status of the hitherto controversial species M. japana (Jacoby, 1885) is confirmed. The pattern of nucleotide and amino acid substitutions is discussed in the light of functional domains of the COI molecule. Although the data are preliminary, the results provide new data on the distribution of the species. Together with the phylogenetic analysis they allow for a discussion of the phylogeography of the genus.

scholarly journals Design of a species-specific PCR method for the detection of the heat-resistant fungi Talaromyces macrosporus and Talaromyces trachyspermus

2017 ◽  
Vol 66 (1) ◽  
pp. 86-92 ◽  
Author(s):  
S. Yamashita ◽  
H. Nakagawa ◽  
T. Sakaguchi ◽  
T-H. Arima ◽  
Y. Kikoku
Keyword(s):  
Heat Resistant ◽  
Specific Pcr ◽  
Pcr Method ◽  
Species Specific

scholarly journals Growth of bifidobacteria in mammalian milk

2013 ◽  
Vol 58 (No. 3) ◽  
pp. 99-105 ◽  
Author(s):  
Š. Ročková ◽  
V. Rada ◽  
J. Havlík ◽  
R. Švejstil ◽  
E. Vlková ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Human Milk ◽  
Bacterial Species ◽  
Intestinal Bacteria ◽  
Bifidobacterium Bifidum ◽  
Microbial Colonization ◽  
Animal Origin ◽  
Good Growth ◽  
Animal Milk ◽  
Species Specific

Microbial colonization of the mammalian intestine begins at birth, when from a sterile state a newborn infant is exposed to an external environment rich in various bacterial species. An important group of intestinal bacteria comprises bifidobacteria. Bifidobacteria represent major intestinal microbiota during the breast-feeding period. Animal milk contains all crucial nutrients for babies’ intestinal microflora. The aim of our work was to test the influence of different mammalian milk on the growth of bifidobacteria. The growth of seven strains of bifidobacteria in human milk, the colostrum of swine, cow’s milk, sheep’s milk, and rabbit’s milk was tested. Good growth accompanied by the production of lactic acid was observed not only in human milk, but also in the other kinds of milk in all three strains of Bifidobacterium bifidum of different origin. Human milk selectively supported the production of lactic acid of human bifidobacterial isolates, especially the Bifidobacterium bifidum species. The promotion of bifidobacteria by milk is species-specific. Human milk contains a key factor for the growth of specific species or strains of human-origin bifidobacteria compared to other kinds of milk. In contrast, some components (maybe lysozyme) of human milk inhibited the growth of Bifidobacterium animalis. Animal-origin strains of bifidobacteria were not able to significantly grow even in milk of animal origin, with the exception of B. animalis subsp. lactis 1,2, which slightly grew in sheep’s milk.

scholarly journals Species level identification of thermotolerant campylobacters 

2012 ◽  
Vol 50 (No. 12) ◽  
pp. 543-547 ◽  
Author(s):  
I. Kolackova ◽  
R. Karpiskova
Keyword(s):  
Species Identification ◽  
Mixed Cultures ◽  
Species Determination ◽  
The Czech Republic ◽  
Identification Process ◽  
Pcr Method ◽  
Phenotypic Methods ◽  
Level Identification ◽  
Campylobacter Spp

The aim of this study was to compare the phenotypic and genotypic based methods for species identification of thermotolerant campylobacters of human and food origin from the Czech Republic. Phenotypic methods are time-consuming and sometimes lead to intermediate results, therefore replacement by more specific and rapid methods are needed. Out of a total of 911 campylobacter strains tested, 800 human isolates were received from the clinical bacteriology laboratories from 5 regions and 111 foodstuff isolates (raw chicken and pork meat from retail market) originated from the routine examination in our laboratory. Based on the PCR method 85.1% of these strains were identified as C. jejuni, 12.5% as C. coli and 2.3% as mixed cultures of C. jejuni and C. coli. When species determination of campylobacters was based on conventional methods (hippurate hydrolysis test), 28.5% of the isolates were not identified correctly. The mixed cultures of campylobacters have not been detected without further subculturing of strains, which takes several days and enormously extends the identification process. The use of the PCR method showed to be a useful tool for species identification of Campylobacter spp.

scholarly journals Isolation and Characterization of Brachyspira spp. from Dogs in the Czech Republic

2010 ◽  
Vol 79 (3) ◽  
pp. 437-442
Author(s):  
Daniel Šperling ◽  
František Čada ◽  
Alois Čížek
Keyword(s):  
Czech Republic ◽  
Biochemical Activity ◽  
Animal Shelters ◽  
The Czech Republic ◽  
Specific Pcr ◽  
Isolation And Characterization ◽  
High Prevalence ◽  
Species Specific

The objectives of this study were to establish the prevalence of intestinal Spirochetes of the genusBrachyspirain Czech dogs and to determine the susceptibility of obtainedB. pilosicoliisolates to selected antibacterial substances. Spirochetes were diagnosed microscopically in 23 out of 1139 samples of dogs’ excrements, primarily intended for a parasitological testing. The cultivation of positive samples provided 10 brachyspira isolates, which were, on the basis of their biochemical activity and the results of the species-specific PCR, identified asB. pilosicoli(9 isolates) andB. hyodysenteriae(1 isolate). These dogs came from households. All the 7 tested isolatesB. pilosicoliwere sensitive to metronidazole and doxycycline, uniformly resistant to erythromycin, partly sensitive to cefazoline, lincomicine and ampicilline except for one isolate ofB. pilosicoli, which was resistant to ampicilline. The second part of study was focused on dogs with diarrhoea that came from animal shelters, where a high prevalence of 58% (10/17) ofB. pilosicoliwas found.

Improved multiplex PCR method for the rapid detection of β-lactamase genes in Escherichia coli of animal origin

2006 ◽  
Vol 56 (1) ◽  
pp. 103-106 ◽  
Author(s):  
Constança Pomba ◽  
Nuno Mendonça ◽  
Marta Costa ◽  
Deolinda Louro ◽  
Bruno Baptista ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Rapid Detection ◽  
Animal Origin ◽  
Pcr Method

Identifying spiders through DNA barcodes

2005 ◽  
Vol 83 (3) ◽  
pp. 481-491 ◽  
Author(s):  
Rowan D.H Barrett ◽  
Paul D.N Hebert
Keyword(s):  
Mating Systems ◽  
Mitochondrial Gene ◽  
Model Systems ◽  
Dna Barcodes ◽  
Cytochrome C Oxidase I ◽  
Accurate Identification ◽  
Spider Species ◽  
Taxonomic Impediment ◽  
Gene Coding ◽  
Identification Tool

With almost 40 000 species, the spiders provide important model systems for studies of sociality, mating systems, and sexual dimorphism. However, work on this group is regularly constrained by difficulties in species identification. DNA-based identification systems represent a promising approach to resolve this taxonomic impediment, but their efficacy has only been tested in a few groups. In this study, we demonstrate that sequence diversity in a standard segment of the mitochondrial gene coding for cytochrome c oxidase I (COI) is highly effective in discriminating spider species. A COI profile containing 168 spider species and 35 other arachnid species correctly assigned 100% of subsequently analyzed specimens to the appropriate species. In addition, we found no overlap between mean nucleotide divergences at the intra- and inter-specific levels. Our results establish the potential of COI as a rapid and accurate identification tool for biodiversity surveys of spiders.

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