Coagulase gene polymorphism and antimicrobial susceptibility of Staphylococcus aureus isolated from bovine subclinical mastitis milk in Sidi-Bel-Abbes, Algeria

2017 ◽  
Vol 7 (1) ◽  
pp. 21-27
Author(s):  
Ahmed Reda Belmamoun ◽  
Abdelkader Ammam ◽  
Imene Berrabah ◽  
Karima Bereksi Reguig
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Pcr Amplification ◽  
Screen Test ◽  
Raw Milk ◽  
Subclinical Mastitis ◽  
Latex Agglutination ◽  
Pcr Analysis ◽  
California Mastitis Test ◽  
Coa Gene

The study was conducted to identify and characterize Staphylococcus aureus in raw milk derived from subclinical mastitis in Sidi-Bel-Abbes Algeria. In this paper, we explore the possibility of detection of the coagulase gene (coa), which encodes the coagulase enzyme, by PCR analysis in antibiotic-resistant isolates, with the latex agglutination phenotype and free coagulase.Out of 336 samples of raw milk examined with California mastitis test (CMT) posi-tive; a total of 142 samples were bacteriologically positive with 56.34% Staphylococcus isolates, 21 (26.25%) isolates were confirmed as S.aureus. Nineteen (90.48%) isolates of S.aureus showed free coagulase on the tube agglutination test. Two atypical S.aureus strains (9.52%) were defective for the clumping factor and / or protein A , determined with the Staphytect plus test and the tube coagulase test. The isolates of S.aureus were resistant to penicillin and tetracycline with 76.19%. Two isolates (9.52%) of S.aureus re-sistant to meticillin (MRSA) were detected in this study, with a MIC of ≥4 μg / liter and a cefoxitin screen test with a MIC of ≥8 μg / liter, and 13 (61.9%) isolates were with a multiresistance phenotype. The 21 isolates were sub-jected to PCR amplification of the 3' end of the coa gene, 18 (85.71%) were revealed on a 1% agarose gel with a single band between 547 bp and 875 bp. The use of the PCR genotypic test to identify the profile of the coa gene can be used as an appropriate identification criterion for differentiating coagulases from S.aureus and for understanding their epidemiology.

Risk from Staphylococcus aureus in informally marketed raw cow milk

2019 ◽  
Vol 2 (1) ◽  
pp. 74 ◽  
Author(s):  
Desislava Bangieva ◽  
Valentin Rusev
Keyword(s):  
Staphylococcus Aureus ◽  
16S Rrna ◽  
Pcr Amplification ◽  
Raw Milk ◽  
Cow Milk ◽  
Pcr Analysis ◽  
Milk Samples ◽  
Staphylococcus Spp ◽  
And Staphylococcus Aureus

This study aimed to investigate the presence of Staphylococcus spp. and Staphylococcus aureus in raw cow milk samples taken from peddlers of five regions in Bulgaria. The results showed that all 44 samples tested were positive for Staphylococcus spp. All isolates were tested for coagulase production and subjected to PCR analysis. PCR amplification of 16S rRNA and nuc genes found the presence of Staphylococcus aureus in 23 (52.3%) of a total of 44 raw milk samples. The number of Staphylococcus spp. ranged from 3x102 to 1.08x106 cfu/ml, and that of Staphylococcus aureus from 1.5x102 to 3.19x105 cfu/ml. It is concluded that control over the hygiene of handling and processing raw milk is essential for its safety.

scholarly journals THE OCCURRENCE OF BACTERIAL AGENTS CAUSING MASTITIS IN DAIRY SHEEP AND THEIR RESISTANCE TO ANTIBIOTICS

2020 ◽  
Vol 3 (7) ◽  
Author(s):  
František Zigo ◽  
Katarína Veszelits Lakticová ◽  
Mária Vargová M
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
High Resistance ◽  
Subclinical Mastitis ◽  
Causative Organism ◽  
Coagulase Negative Staphylococci ◽  
Isolation And Identification ◽  
Milk Samples ◽  
California Mastitis Test ◽  
Acute Form

Introduction: The aim of this study was to monitor the occurrence of bacterial agents causing mastitis, forms of mastitis and antibiotic resistance in 300 ewes on a farm in Eastern Slovakia. Material and methods: During the milking season, were performed three complex investigations including clinical examination, California Mastitis Test and laboratory analysis of milk samples. The investigations and milk samples were taken in three phases; the beginning (April), the middle (June) and the end (September) of the milking season. Results: Of all the samples (806), 225 (28.0%) were positive for bacterial pathogens. The highest incidence of mastitis (33.3%) was recorded in September, while April (23.8%) and June (25.3%) had lower incidence. The samples from September showed the highest incidence of subclinical mastitis (20.1%), with 13.2% being an acute form of mastitis. Coagulase-negative staphylococci (CNS) were identified in 61.7% of the positive samples. Especially, S. chromogenes, S. epidermidis and S. schleiferi were most frequently isolated.  Staphylococcus aureus was the causative agent in 20.0% of the positive samples and caused acute or subclinical mastitis in the affected ewes. The tested bacteria showed very high resistance to novobiocin (59.5%) and penicillin (51.4%) and high resistance to amoxycillin (35.1%). We found that 80% of Staphylococcus aureus bacteria tested for antibiotic resistance were resistant to novobiocin and 70% were resistant to penicillin. Of all tested CNS, 56.5% were resistant to novobiocin, 39.1% to penicillin, and 34.7% to amoxicillin. Conclusion: Proper isolation and identification of the causative organism play a significant role in the prevention and control of the intramammary infection. In our study, a combinations of Streptomycin, Ciprofloxacin and Tetracycline were the most effective antibiotics for the control of mastitis.

scholarly journals CidR and CcpA Synergistically Regulate Staphylococcus aureus cidABC Expression

2019 ◽  
Vol 201 (23) ◽  
Author(s):  
Marat R. Sadykov ◽  
Ian H. Windham ◽  
Todd J. Widhelm ◽  
Vijaya Kumar Yajjala ◽  
Sean M. Watson ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Carbon Metabolism ◽  
Protein A ◽  
Transcriptional Regulators ◽  
Extracellular Dna ◽  
Regulatory Control ◽  
Pcr Analysis ◽  
Mobility Shift ◽  
Content Type ◽  
Biofilm Development

ABSTRACT The death and lysis of a subpopulation of Staphylococcus aureus cells during biofilm development benefit the whole bacterial population through the release of an important component of the biofilm matrix, extracellular DNA. Previously, we have demonstrated that these processes are affected by the gene products of the cidABC operon, the expression of which is controlled by the LysR-type transcriptional regulator, CidR. In this study, we characterized cis- and trans-acting elements essential for the induction of the cidABC operon. In addition to a CidR-binding site located within the cidABC promoter region, sequence analysis revealed the presence of a putative catabolite responsive element (cre box), suggestive of the involvement of the catabolite control protein A (CcpA) in the regulation of cidABC expression. This was confirmed using electrophoretic mobility shift assays and real-time reverse transcriptase PCR analysis demonstrating the direct positive control of cidABC transcription by the master regulator of carbon metabolism. Furthermore, the importance of CcpA and the identified cre site for the induction of the cidABC operon was demonstrated by examining the expression of PcidABC-lacZ reporter fusions in various mutant strains in which the genes involved in carbon metabolism and carbon catabolite repression were disrupted. Together the results of this study demonstrate the necessity of both transcriptional regulators, CidR and CcpA, for the induction of the cidABC operon and reveal the complexity of molecular interactions controlling its expression. IMPORTANCE This work focuses on the characterization of cis- and trans-acting elements essential for the induction of the cidABC operon in S. aureus. The results of this study are the first to demonstrate the synergistic control of cidABC expression by transcriptional regulators CidR and CcpA during carbohydrate metabolism. We established that the full induction of cidABC expression depends on the metabolic state of bacteria and requires both CidR and CcpA. Together, these findings delineate regulatory control of cidABC expression under different metabolic conditions and provide important new insights into our understanding of cell death mechanisms during biofilm development in S. aureus.

Characterization of Borderline Oxacillin-Resistant Staphylococcus aureus Isolated from Food of Animal Origin

2010 ◽  
Vol 73 (7) ◽  
pp. 1325-1327 ◽  
Author(s):  
JAROSŁAW BYSTROŃ ◽  
MAGDALENA PODKOWIK ◽  
KAMILA KORZEKWA ◽  
ELŻBIETA LIS ◽  
JERZY MOLENDA ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protein A ◽  
Meca Gene ◽  
Raw Milk ◽  
Animal Origin ◽  
Molecular Characteristics ◽  
Type I ◽  
Staphylococcal Protein A ◽  
Food Of Animal Origin ◽  
Resistant Strains

In this study, the molecular characteristics of food-derived oxacillin-resistant Staphylococcus aureus were determined. Eight borderline oxacillin-resistant strains with MICs of 2 to 4 μg/ml were identified from 132 S. aureus isolates of food origin. One of the two isolates with a MIC of 4 μg/ml was methicillin-resistant determinant (mecA) gene positive, and the other six with MICs of 2 μg/ml were mecA negative. The mecA-positive isolate was classified as sequence type (ST)228, staphylococcal protein A (spa) type t041, and carried the staphylococcal cassette chromosome mec type I element. Two borderline oxacillin-resistant strains were classified as spa t008 and ST8, and the remaining five as spa t164 and ST20. The mecA-positive strain and four borderline oxacillin-resistant strains were found enterotoxigenic. The enterotoxin genes detected in these strains included selp, egc1, and sed-sej-selr. The borderline-resistant S. aureus isolates from a manually handled product, i.e., minced pork, were shown genetically related to strains associated with human infections. This suggests that humans can be considered as a source of contamination of this food with oxacillin-resistant S. aureus strains. The genotypes of the investigated milk borderline-resistant isolates were shown to occur not only in cows, but also in humans. Since manual handling is reduced in raw milk production, a human origin of S. aureus seems unlikely. Because knowledge of the genotypes of animal staphylococci is limited, more research is needed to address the question of the origin of antibiotic-resistant S. aureus strains in food.

Staphylococcus aureus Isolates from Irish Domestic Refrigerators Possess Novel Enterotoxin and Enterotoxin-like Genes and Are Clonal in Nature

2006 ◽  
Vol 69 (3) ◽  
pp. 508-515 ◽  
Author(s):  
DAVIDA S. SMYTH ◽  
JEAN KENNEDY ◽  
JANE TWOHIG ◽  
HELEN MIAJLOVIĆ ◽  
DECLAN BOLTON ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Broiler Chickens ◽  
Random Amplified Polymorphic Dna ◽  
Protein A ◽  
Food Poisoning ◽  
Staphylococcal Enterotoxin ◽  
Penicillin G ◽  
Animal Origin ◽  
Pcr Analysis ◽  
Potential Reservoir

A previous study carried out by the National Food Centre in Dublin on bacterial contamination of Irish domestic refrigeration systems revealed that 41% were contaminated with Staphylococcus aureus. One hundred fifty-seven S. aureus isolates were screened by multiplex PCR analysis for the presence of 15 staphylococcal enterotoxin and enterotoxin-like genes (seasee, seg-sei, selj-selo, and selq) and the toxic shock toxin superantigen tst gene. Of the refrigerator isolates, 64.3% possessed more than one staphylococcal enterotoxin or staphylococcal enterotoxin–like gene. All bar one of the 101 staphylococcal enterotoxin or staphylococcal enterotoxin–like gene-positive strains possessed the egc locus bearing the seg, sei, selm, seln, and selo genes. Twelve random amplified polymorphic DNA (RAPD) types accounted for 119 (75.8%) of the strains, two of these types accounting for 25 (RAPD type 1, 15.9%) and 52 (RAPD type 5, 33.1%), respectively. All of the RAPD type 5 isolates possessed the egc gene cluster only. The RAPD type 5 amplicon profile was identical to that of S. aureus isolates associated with osteomyelitis in broiler chickens in Northern Ireland that also possessed the egc locus only. However, the RAPD type 5 domestic refrigerator and chicken isolates differed in penicillin G sensitivity, production of Protein A and staphylokinase, and crystal violet agar growth type. These findings highlight that the average Irish household refrigerator harbors potential enterotoxin-producing S. aureus that may or may not be of animal origin and, accordingly, is a potential reservoir for staphylococcal food poisoning.

scholarly journals Preliminary study of homeopathic treatment of subclinical mastitis evaluated through somatic cells count (SCC) and California mastitis test (CMT)

2021 ◽  
Vol 7 (24) ◽  
pp. 147-151
Author(s):  
Zilda Cristiani Gazim ◽  
Orlando Seiko Takemura ◽  
Luiz Cláudio Monteiro da Silva ◽  
Carlos Massambani ◽  
Roberson Eduardo Moraes ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Somatic Cells ◽  
Subclinical Mastitis ◽  
Clinical Aspects ◽  
Infection Level ◽  
Homeopathic Treatment ◽  
Holstein Breed ◽  
California Mastitis Test ◽  
End Of Treatment ◽  
Preliminary Study

The present paper presents the results of homeopathic treatment of 25 Holstein breed cows aged 3 to 8 years old diagnosed with subclinical mastitis through California Mastitis Test (CMT). Animals were divided into 3 groups according with infection level. A homeopathic complex was developed on the grounds of clinical aspects, including Phosphorus 30x, Phytolacca 30x, Silicea 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Pulsatilla 30x, Calendula 30x and biotherapic of Staphylococcus aureus 200x. The remedy was added to salt and was administered to cattle 100g/cow/day for 75 days. CMT were carried out every 2 weeks to control incidence and severity of mastitis; somatic cells count (SCC) was performed at the beginning and the end of treatment. CMT showed significant improvement in regression of infection level all throughout the study; final SCC showed decrease in 82% of animals, signaling thus efficacy of the homeopathic treatment. Keywords:  Subclinical mastitis; Dairy cows; Homeopathy; California Mastitis Test, Somatic Cells Count.   Estudo preliminar do tratamento homeopático da mastite subclínica avaliado por contagem de células somáticas (SCC) e California Mastitis Test (CMT) Resumo Este artigo apresenta os resultados do tratamento homeopático de 25 vacas raça Holstein entre 3 e 8 anos de idade diagnosticadas com mastite subclínica através do California Mastitis Test (CMT). Os animais foram divididos em 3 grupos de acordo com o nível da infecção. Foi preparado um complexo homeopático com base nos achados clínicos, composto de: Phosphorus 30X, Phytolacca 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Silicea 30x, Pulsatilla 30x, Calendula 30x s 30X, Phytolacca 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Silicea 30x, Pulsatilla 30x, Calendula 30x e bioterápico de Staphylococcus aureus 200x. O complexo foi acrescentado no sal e administrado na dose de 100g/vaca/dia. O CMT foi realizado a cada 2 semanas a fim de monitorar a incidéncia e gravidade da mastitie; a contagem de células somáticas (SCC) foi realizada ao início e no final do tratamento. Os valores do CMT mostraram melhora significativa no sentido de regressão do nível da infecção ao longo do estudo; o valor final da SCC diminuiu em 82% dos animais, apontando para a eficácia do tratamento homeopático. Palavras-chave:  Mastite subclínica; Vacas leiteiras; Homeopatia; CMT, contagem de células somáticas.   Estudio preliminar del tratamiento homeopático de la mastitis sublínica evaluado mediante recuento de células somáticas (SCC) y California Mastitis Test (CMT) Resumen Este artículo presenta los resultados del tratamiento homeopático de 25 vacas raza Holstein entre 3 e 8 años de edad diagnosticadas com mastitis sucblínica mediante California Mastitis Test (CMT). Los animales fueron divididos en 3 grupos según el nível de la infección. Fue compuesto un complejo homeopático con base en la presentación clínica, incluyendo Phosphorus 30X, Phytolacca 30x, Sulphur 30x, Belladona 30x, Bryonia alba 30x, Silicea 30x, Pulsatilla 30x, Calendula 30x y bioterápico de Staphylococcus aureus 200x. El complejo fue agregado en sal y administrado a los animales en la dosis de 100g/vaca/día. CMT fue realizado cada 2 semanas para monitorizar la incidencia y gravedad de la mastitis; recuento de células somáticas (SCC) fue realizado al inicio y al final del tratamiento. CMT mostró mejoría significativa en el sentido de regresión del nivel de la infección a lo largo del estudio; el valor final del SCC disminuyó en 82% de los animales, apuntando para la eficacia del tratamiento homeopático. Palabras-clave:  Mastitis subclínica; Vacas lecheras; Homeopatía; CMT, recuento de células somáticas.   Correspondence author:  Zilda Cristiani Gazim, [email protected] How to cite this article:  Barzon CD, Medeiros F, Moraes RE, Silva LCM, Massambani C, Takemura OS, Gazim ZC. Preliminary study of homeopathic treatment of subclinical mastitis evaluated through somatic cells count and California mastitis test. Int J High Dilution Res [online]. 2008 [cited YYYY Mmm DD]; 7(24): 147-151. Available from: http://journal.giri-society.org/index.php/ijhdr/article/view/292/364.  

scholarly journals ЕТІОЛОГІЧНІ ЧИННИКИ МАСТИТІВ КОРІВ УКРАЇНСЬКОЇ ЧОРНО–РЯБОЇ МОЛОЧНОЇ ПОРОДИ

2016 ◽  
Vol 18 (3(70)) ◽  
pp. 191-196
Author(s):  
V. Panevnyk ◽  
T. Suprovych
Keyword(s):  
Staphylococcus Aureus ◽  
Somatic Cells ◽  
Raw Milk ◽  
Subclinical Mastitis ◽  
Index Number ◽  
Clinical Form ◽  
Leading Role ◽  
Skin Contamination ◽  
Major Pathogens ◽  
And Staphylococcus Aureus

The article shows data on the microbial landscape and quantity of somatic cells milk in different forms of mastitis in cows. Index number of somatic cells (SCC) in the raw milk of cows in the country is only used to establish the quality milk. They are key safety indicators that are directly related to udder cow disease, especially subclinical mastitis. Research has established that the number of SCC in healthy animals ranges from 84000 cells/ml to 436000 cells/ml. Over the course of subclinical turned from 508000 cells/ml to 756000 cells/ml. Animals with clinical form of mastitis were from 876000 cells/ml to 69260000 cells/ml. The 42 cows of the first lactation average SCC was 143000 cells/ml, and 47 of fifth lactation cows – 213000 cells/ml.The microflora in the breast can get in different ways: galactogenous – through teat channel hematogenous ahd lymphogenous ways. The leading role galactogenous way in which the pathogens penetrate from the environment through teat channel. This contributes to the udder skin contamination by microorganisms. Activators of subclinical mastitis were Staphylococcus aureus 31.8% and Streptococcus agalactiae 40.9%. In the clinical course of mastitis major pathogens were Escherichia coli – 34.8% and Staphylococcus aureus – 41.3%. Selected cultures of microorganisms were sensitive to cephalexin, gentamicin, oxacillin, rifampicin, enrofloksacin. 

MOLECULAR TYPING OF METHICILLIN-SUSCEPTIBLE STAPHYLOCOCCUS AUREUS (MSSA), ISOLATED FROM MONKEYS, BASED ON COAGULASE GENE POLYMORPHISM

2019 ◽  
pp. 57-62
Author(s):  
V. A. Kalashnikova
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Molecular Genetic ◽  
Bovine Mastitis ◽  
Variable Region ◽  
Staphylococcal Infection ◽  
Pcr Analysis ◽  
Polymorphism Analysis ◽  
Coa Gene

Staphylococcus aureus is a very dangerous microorganism that causes more than 100 nosological forms of disease in humans and animals, including pneumonia, skin and soft tissue infections, food toxicoinfections, wound abscess, etc. Numerous studies on genotyping Staphylococcus aureus, isolated from humans, food and bovine mastitis have been carried out. The lack of information on the genotyping of these pathogens detected in monkeys living in captivity served as a stimulus to conduct a similar research, since staphylococcal infections in the primates are widespread. The present study is devoted to the study of the polymorphism of a variable region of the coagulase gene and to the typing of Staphylococcus aureus isolates from monkeys of different species kept at Adler monkey farm. 115 Staphylococcus aureus isolates were studied using phenotypic and molecular genetic methods. Genotyping was performed using PCR, real-time PCR and PCR with subsequent restriction fragment length polymorphism analysis (PCR-RFLP). A real-time PCR analysis allowed to classify all Staphylococcus aureus as methicillin-susceptible staphylococci (MSSA). After amplification of a variable region of the coagulase gene, 4 types of amplicons of 600, 700, 800, and 900 bp were generated. This data demonstrates structural differences of this gene in the studied isolates. The coagulase gene of 900 bp prevailed. The use of the Cfo1 endonuclease allowed to identify 23 different restriction profiles of the coa gene, but only three of them predominated. Staphylococcus aureus bacteria with seven types of coagulase gene were found only in the lungs of monkeys that died of pneumonia. The results obtained suggest that these isolates have tropism for lung tissue. Among Staphylococcus aureus isolated from pneumonia cases, isolates with three types of the coa gene prevailed. Staphylococcus aureus of eleven types of coagulase gene can be attributed to the invasive isolates, since they were detected in the tissues of various organs. Staphylococcal infection in monkeys kept at the monkey farm is caused by genotypically heterogeneous population of Staphylococcus aureus.

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