scholarly journals Phenotypic and Genotypic Characterization ofStaphylococcus aureusStrains from Italian Dairy Products

2009 ◽  
Vol 2009 ◽  
pp. 1-7 ◽  
Author(s):  
Stefano Morandi ◽  
Milena Brasca ◽  
Cristian Andrighetto ◽  
Angiolella Lombardi ◽  
Roberta Lodi
Keyword(s):  
Dairy Products ◽  
Animal Species ◽  
Randomly Amplified Polymorphic Dna ◽  
Foodborne Illnesses ◽  
Phenotypic Properties ◽  
Rapd Pcr ◽  
Genes Encoding ◽  
Dna Pcr ◽  
Phenotypic And Genotypic Characterization ◽  
Milk And Dairy Products

Staphylococcus aureusis a known major cause of foodborne illnesses, and milk and dairy products are often contaminated by enterotoxigenic strains of this bacterium. In the present study, 122S. aureusisolates collected from different dairy products were characterised by phenotypic properties, by the distribution of genes encoding staphylococcal enterotoxins (sea,sec,sed,seg,seh,sei,sej, andsel) and by randomly amplified polymorphic DNA PCR (RAPD-PCR). Moreover, strain resistance to vancomycin and methicillin (oxacillin) was studied. The differences in the RAPD-PCR profiles obtained with the primers M13 and AP4 revealed the presence of a great genetic heterogeneity among the differentS. aureusstrains. Using the primer AP4 and M13, eight groups were distinguished by RAPD-PCR cluster analysis, although, except in few cases, it was not possible to correlate the isolates of different animal species (cow or ovine) with the presence ofsegenes. None of the isolates showed resistance to vancomycin or methicillin.

Use of PCR-Based Methods for Rapid Differentiation of Lactobacillus delbrueckii subsp.bulgaricus and L. delbrueckii subsp.lactis

1999 ◽  
Vol 65 (10) ◽  
pp. 4351-4356 ◽  
Author(s):  
Sandra Torriani ◽  
Giacomo Zapparoli ◽  
Franco Dellaglio
Keyword(s):  
Numerical Analysis ◽  
Dairy Products ◽  
Lactobacillus Delbrueckii ◽  
Randomly Amplified Polymorphic Dna ◽  
Specific Pcr ◽  
Single Colony ◽  
Rapd Pcr ◽  
Reliable Differentiation ◽  
Dna Pcr ◽  
Proline Iminopeptidase

ABSTRACT Two PCR-based methods, specific PCR and randomly amplified polymorphic DNA PCR (RAPD-PCR), were used for rapid and reliable differentiation of Lactobacillus delbrueckii subsp.bulgaricus and L. delbrueckii subsp.lactis. PCR with a single combination of primers which targeted the proline iminopeptidase (pepIP) gene ofL. delbrueckii subsp. bulgaricus allowed amplification of genomic fragments specific for the two subspecies when either DNA from a single colony or cells extracted from dairy products were used. A numerical analysis of the RAPD-PCR patterns obtained with primer M13 gave results that were consistent with the results of specific PCR for all strains except L. delbrueckii subsp.delbrueckii LMG 6412T, which clustered withL. delbrueckii subsp. lactis strains. In addition, RAPD-PCR performed with primer 1254 provided highly polymorphic profiles and thus was superior for distinguishing individual L. delbrueckii strains.

scholarly journals A Pseudomonas syringae Diversity Survey Reveals a Differentiated Phylotype of the Pathovar syringae Associated with the Mango Host and Mangotoxin Production

2013 ◽  
Vol 103 (11) ◽  
pp. 1115-1129 ◽  
Author(s):  
José A. Gutiérrez-Barranquero ◽  
Víctor J. Carrión ◽  
Jesús Murillo ◽  
Eva Arrebola ◽  
Dawn L. Arnold ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Catabolic Diversity ◽  
Rapd Pcr ◽  
Geographical Regions ◽  
Extensive Collection ◽  
Polymerase Chain ◽  
Apical Necrosis ◽  
Dna Pcr

Pseudomonas syringae pv. syringae, the causal agent of bacterial apical necrosis (BAN) in mango crops, has been isolated in different mango-producing areas worldwide. An extensive collection of 87 P. syringae pv. syringae strains isolated from mango trees affected by BAN from different countries, but mainly from Southern Spain, were initially examined by repetitive sequence-based polymerase chain reaction (rep-PCR) to analyze the genetic diversity with an epidemiological aim. rep-PCR was powerful in assessing intrapathovar distribution and also allowing clustering of the P. syringae pv. syringae strains isolated from mango, depending on the isolation area. A clear pattern of clustering was observed for all the P. syringae pv. syringae strains isolated from mango distinct from strains from other hosts, including strains for the same geographical regions as the mango isolates. For this reason, a representative group of 51 P. syringae pv. syringae strains isolated from mango and other hosts, as well as some P. syringae strains from other pathovars, were further characterized to determine their possible genetic, phenotypic, and phylogenetic relationships. Similar to the rep-PCR results, the randomly amplified polymorphic DNA PCR (RAPD-PCR) and catabolic diversity analysis using the Biolog GN2 profile grouped 90% of the mango isolates together in a unique cluster. Interestingly, the majority of P. syringae pv. syringae strains isolated from mango produced mangotoxin. The analysis of the phylogenetic distribution using the multilocus sequence typing analysis strongly supports the existence of a differentiated phylotype of the pathovar syringae mainly associated with the mango host and characterized by the mangotoxin production.

scholarly journals Seasonal Dynamics and Metagenomic Characterization of Estuarine Viriobenthos Assemblages by Randomly Amplified Polymorphic DNA PCR

2009 ◽  
Vol 75 (8) ◽  
pp. 2259-2265 ◽  
Author(s):  
Rebekah R. Helton ◽  
K. Eric Wommack
Keyword(s):  
Chesapeake Bay ◽  
Oligonucleotide Primer ◽  
Viral Diversity ◽  
Randomly Amplified Polymorphic Dna ◽  
Aquatic Sediments ◽  
Banding Patterns ◽  
Rapd Pcr ◽  
Viral Sequences ◽  
Dna Pcr

ABSTRACT Direct enumeration and genetic analyses indicate that aquatic sediments harbor abundant and diverse viral communities. Thus far, synecological analysis of estuarine sediment viral diversity over an annual cycle has not been reported. This oversight is due in large part to a lack of molecular genetic approaches for assessing viral diversity within a large collection of environmental samples. Here, randomly amplified polymorphic DNA PCR (RAPD-PCR) was used to examine viral genotypic diversity within Chesapeake Bay sediments. Using a single 10-mer oligonucleotide primer for all samples, RAPD-PCR analysis of sediment viral assemblages yielded unique banding patterns across spatial and temporal scales, with the occurrence of specific bands varying among the sample set. Cluster analysis of RAPD-PCR amplicon banding patterns indicated that sediment viral assemblages changed with season and to a lesser extent with geographic location. Sequence analysis of RAPD-PCR amplicons revealed that 76% of sediment viral sequences were not homologous to any sequence in the GenBank nonredundant protein database. Of the GenBank sequence homologs, the majority belonged to viruses within the Podoviridae (24%) and Myoviridae (22%) viral families, which agrees with the previously observed frequencies of these morphological families in Chesapeake Bay sediments. Furthermore, the majority of the sediment viral sequences homologous to GenBank nonredundant protein sequences were phages or prophages (57%). Hence, RAPD-PCR proved to be a reliable and useful approach for characterization of viral assemblages and the genetic diversity of viruses within aquatic sediments.

Randomly amplified polymorphic DNA (RAPD) PCR for the identification of yeasts isolated from dairy products

2000 ◽  
Vol 30 (1) ◽  
pp. 5-9 ◽  
Author(s):  
C. Andrighetto ◽  
E. Psomas ◽  
N. Tzanetakis ◽  
G. Suzzi ◽  
A. Lombardi
Keyword(s):  
Dairy Products ◽  
Randomly Amplified Polymorphic Dna ◽  
Rapd Pcr

Differentiation of faecal Escherichia coli from human and animal sources by random amplified polymorphic DNA-PCR (RAPD-PCR)

2004 ◽  
Vol 50 (1) ◽  
pp. 193-198 ◽  
Author(s):  
D. Venieri ◽  
A. Vantarakis ◽  
G. Komninou ◽  
M. Papapetropoulou
Keyword(s):  
Escherichia Coli ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Hierarchical Cluster ◽  
Group Method ◽  
Randomly Amplified Polymorphic Dna ◽  
Arbitrary Primers ◽  
E Coli ◽  
Rapd Pcr ◽  
Dna Pcr

In this study the assessment of randomly amplified polymorphic DNA (RAPD) analysis was established as a molecular epidemiological tool. RAPD analysis was performed to differentiate faecal Escherichia coli isolates from human and animal sources. E. coli strains (128) were isolated from human and animal faeces (from cattle and sheep). Genomic DNA was extracted and randomly amplified polymorphic DNA-PCR (RAPD-PCR) fingerprinting was performed. Seven arbitrary primers were tested with a view to discriminating between E. coli isolates from humans and E. coli isolates from animals. RAPD profiles were analysed with hierarchical cluster analysis using an unweighted pair group method. RAPD profiles obtained with three of the tested primers (1247, 1290 and 1254) established a distinct differentiation between E. coli isolates from humans and E. coli from animals. Low levels of misclassification and high levels of specificity make RAPD a sensitive, efficient and reliable means of distinguishing closely related strains.

Occurrence of Enterotoxic Staphylococcus aureus in Raw Milk from Yaks and Cattle in Mongolia

2007 ◽  
Vol 70 (7) ◽  
pp. 1726-1729 ◽  
Author(s):  
URANCHIMEG TSEGMED ◽  
GIOVANNI NORMANNO ◽  
MARIT PRINGLE ◽  
KAREL KROVACEK
Keyword(s):  
Staphylococcus Aureus ◽  
Dairy Products ◽  
Food Poisoning ◽  
Raw Milk ◽  
Latex Agglutination ◽  
Animal Origin ◽  
Foodborne Illnesses ◽  
First Report ◽  
Milk Samples ◽  
Milk And Dairy Products

Staphylococcal food poisoning is considered one of the leading foodborne illnesses in humans worldwide and is associated with contaminated foods of animal origin, such as milk and dairy products. In this study, we investigated the occurrence of staphylococci and the enterotoxigenic properties of Staphylococcus aureus isolated from raw milk from yaks (Bos mutus) and cattle in Mongolia. Staphylococci were isolated from 72 (74%) of the 97 raw milk samples. Of the samples containing staphylococci, 69% (50 of 72) were from yaks and 30.5% (22 of 72) were from cattle. S. aureus was detected in 10% of yak (7 of 72) and 21% of cattle (15 of 72) milk samples. Staphylococcal enterotoxin C was detected in 23% (5 of 22) of the S. aureus strains investigated, based on the reverse passive latex agglutination technique. Three of the five enterotoxigenic strains were from yaks and two were from cattle. None of the S. aureus strains tested produced staphylococcal enterotoxins A, B, or D. To our knowledge, this is the first report of the occurrence of staphylococci and enterotoxigenic S. aureus in milk from yaks and cattle in Mongolia.

scholarly journals Microbial agents in macroscopically healthy mammary gland tissues of small ruminants

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3994
Author(s):  
Liliana Spuria ◽  
Elena Biasibetti ◽  
Donal Bisanzio ◽  
Ilaria Biasato ◽  
Daniele De Meneghi ◽  
...  
Keyword(s):  
Dairy Products ◽  
Lower Risk ◽  
Animal Species ◽  
Multinomial Logistic Regression ◽  
Small Ruminants ◽  
Mammary Glands ◽  
Coagulase Negative Staphylococci ◽  
Microbial Agents ◽  
Microbial Isolates ◽  
Milk And Dairy Products

BackgroundHealth of mammary glands is fundamental for milk and dairy products hygiene and quality, with huge impacts on consumers welfare.MethodsThis study aims to investigate the microbial agents (bacteria, fungi and lentiviruses) isolated from 89 macroscopically healthy udders of regularly slaughtered small ruminants (41 sheep, 48 goats), also correlating their presence with the histological findings. Multinomial logistic regression was applied to evaluate the association between lesions and positivity for different microbial isolates, animal age and bacteria.ResultsTwenty-five samples were microbiologically negative; 138 different bacteria were isolated in 64 positive udders. Coagulase-negative staphylococci were the most prevalent bacteria isolated (46.42%), followed by environmental opportunists (34.76%), others (10.14%) and pathogens (8.68%). Most mammary glands showed coinfections (75%). Lentiviruses were detected in 39.3% of samples. Histologically, chronic non-suppurative mastitis was observed in 45/89 glands, followed by chronic mixed mastitis (12/89) and acute suppurative mastitis (4/89). Only 28 udders were normal. Histological lesions were significantly associated with the animal species and lentiviruses and coagulase-negative staphylococci infections. Goats had significantly higher risk to show chronic mixed mastitis compared to sheep. Goats showed a significantly lower risk (OR = 0.26; 95% CI [0.06–0.71]) of being infected by environmental opportunists compared to sheep, but higher risk (OR = 10.87; 95% CI [3.69–37.77]) of being infected with lentiviruses.DiscussionThe results of the present study suggest that macroscopically healthy glands of small ruminants could act as a reservoir of microbial agents for susceptible animals, representing a potential risk factor for the widespread of acute or chronic infection in the flock.

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