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3.
Rev Assoc Med Bras (1992) ; 68(9): 1191-1198, 2022 Sep.
Article in English | MEDLINE | ID: mdl-36228250

ABSTRACT

OBJECTIVE: Bronchiectasis is a chronic respiratory disease characterized by inflammation, irreversible dilation of the bronchi, and recurrent pulmonary infections, with a high morbidity and mortality rate, but is less studied from the point of view of its prevalence and associated factors not directly related to respiratory prognosis. As it is a disease related to the exacerbation of the inflammatory process and oxidative stress, this study searched to investigate the micronucleus frequency in patients with and without bronchiectasis treated at a specialized pulmonology service in a hospital in the extreme south of Brazil. METHODS: Patients with a confirmed tomographic diagnosis of bronchiectasis were defined as cases. Mutagenicity was evaluated by the micronucleus test in patients' oral mucosa cells. Data collection was performed through a questionnaire containing socioeconomic, demographic, lifestyle, and health condition information. RESULTS: Of the 95 patients involved in this study, 21 (22.1%) were diagnosed with bronchiectasis aged between 12 and 89 years. There was no significant difference in the frequency of micronucleus between patients with and without bronchiectasis. There was a significant positive association between age and frequency of micronucleus among patients with bronchiectasis, but this association does not occur among patients without the disease. CONCLUSION: This is the first study to investigate data on the prevalence and clinical and epidemiological aspects of this chronic disease in Brazil, especially those related to the genotoxicity outcome.


Subject(s)
Bronchiectasis , Pulmonary Medicine , Adolescent , Adult , Aged , Aged, 80 and over , Brazil/epidemiology , Bronchiectasis/diagnosis , Bronchiectasis/epidemiology , Child , Hospitals , Humans , Middle Aged , Mutagens/therapeutic use , Young Adult
4.
Rev. Assoc. Med. Bras. (1992, Impr.) ; Rev. Assoc. Med. Bras. (1992, Impr.);68(9): 1191-1198, Sept. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1406646

ABSTRACT

SUMMARY OBJECTIVE: Bronchiectasis is a chronic respiratory disease characterized by inflammation, irreversible dilation of the bronchi, and recurrent pulmonary infections, with a high morbidity and mortality rate, but is less studied from the point of view of its prevalence and associated factors not directly related to respiratory prognosis. As it is a disease related to the exacerbation of the inflammatory process and oxidative stress, this study searched to investigate the micronucleus frequency in patients with and without bronchiectasis treated at a specialized pulmonology service in a hospital in the extreme south of Brazil. METHODS: Patients with a confirmed tomographic diagnosis of bronchiectasis were defined as cases. Mutagenicity was evaluated by the micronucleus test in patients' oral mucosa cells. Data collection was performed through a questionnaire containing socioeconomic, demographic, lifestyle, and health condition information. RESULTS: Of the 95 patients involved in this study, 21 (22.1%) were diagnosed with bronchiectasis aged between 12 and 89 years. There was no significant difference in the frequency of micronucleus between patients with and without bronchiectasis. There was a significant positive association between age and frequency of micronucleus among patients with bronchiectasis, but this association does not occur among patients without the disease. CONCLUSION: This is the first study to investigate data on the prevalence and clinical and epidemiological aspects of this chronic disease in Brazil, especially those related to the genotoxicity outcome.

5.
Antibiotics (Basel) ; 10(7)2021 Jul 20.
Article in English | MEDLINE | ID: mdl-34356800

ABSTRACT

Staphylococcus aureus and coagulase-negative staphylococci (CoNS) have become the main causative agents of medical device-related infections due to their biofilm-forming capability, which protects them from the host's immune system and from the action of antimicrobials. This study evaluated the ability of RNA III inhibiting peptide (RIP) to inhibit biofilm formation in 10 strains isolated from clinical materials, including one S. aureus strain, two S. epidermidis, two S. haemolyticus, two S. lugdunensis, and one isolate each of the following species: S. warneri, S. hominis, and S. saprophyticus. The isolates were selected from a total of 200 strains evaluated regarding phenotypic biofilm production and the presence and expression of the ica operon. The isolates were cultured in trypticase soy broth with 2% glucose in 96-well polystyrene plates containing catheter segments in the presence and absence of RIP. The catheter segments were observed by scanning electron microscopy. The results showed inhibition of biofilm formation in the presence of RIP in all CoNS isolates; however, RIP did not interfere with biofilm formation by S. aureus. RIP is a promising tool that might be used in the future for the prevention of biofilm-related infections caused by CoNS.

6.
Microb Drug Resist ; 27(6): 823-834, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33232626

ABSTRACT

Bronchiectasis, which is an abnormal and irreversible dilation of one or several bronchial segments, causes significant morbidity and impaired quality of life to patients, mainly as the result of recurrent and chronic respiratory infections. Staphylococcus aureus is a microorganism known for its high infectious potential related to the production of molecules with great pathogenic power, such as enzymes, toxins, adhesins, and biofilm, which determine the degree of severity of systemic symptoms and can induce exacerbated immune response. This review highlighted the clinical significance of S. aureus colonization/infection in bronchiectasis patients, since little is known about it, despite its increasing frequency of isolation and potential serious morbidity.


Subject(s)
Bronchiectasis/complications , Staphylococcal Infections/etiology , Staphylococcus aureus/physiology , Adhesins, Escherichia coli/genetics , Adhesins, Escherichia coli/metabolism , Bacterial Toxins/metabolism , Biofilms/growth & development , Bronchiectasis/mortality , Exotoxins/metabolism , Humans , Leukocidins/metabolism , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/physiology , Microbiota/physiology , Prognosis , Staphylococcus aureus/genetics , Superantigens/immunology
7.
s.l; s.n; 2021. 1-13 p. ilus, tab.
Non-conventional in English | CONASS, Sec. Est. Saúde SP, SESSP-ILSLPROD, Sec. Est. Saúde SP, SESSP-ILSLACERVO, Sec. Est. Saúde SP | ID: biblio-1358675

ABSTRACT

Staphylococcus aureus and coagulase-negative staphylococci (CoNS) have become the main causative agents of medical device-related infections due to their biofilm-forming capability, which protects them from the host's immune system and from the action of antimicrobials. This study evaluated the ability of RNA III inhibiting peptide (RIP) to inhibit biofilm formation in 10 strains isolated from clinical materials, including one S. aureus strain, two S. epidermidis, two S. haemolyticus, two S. lugdunensis, and one isolate each of the following species: S. warneri, S. hominis, and S. saprophyticus. The isolates were selected from a total of 200 strains evaluated regarding phenotypic biofilm production and the presence and expression of the ica operon. The isolates were cultured in trypticase soy broth with 2% glucose in 96-well polystyrene plates containing catheter segments in the presence and absence of RIP. The catheter segments were observed by scanning electron microscopy. The results showed inhibition of biofilm formation in the presence of RIP in all CoNS isolates; however, RIP did not interfere with biofilm formation by S. aureus. RIP is a promising tool that might be used in the future for the prevention of biofilm-related infections caused by CoNS.


Subject(s)
Staphylococcus aureus/isolation & purification , Biofilms , Peptides , RNA , Microscopy, Electron, Scanning , Coagulase , Catheters
8.
Braz J Infect Dis ; 24(2): 160-169, 2020.
Article in English | MEDLINE | ID: mdl-32084346

ABSTRACT

The increasing rates of nosocomial infection associated with coagulase-negative staphylococci (CoNS) were the rationale for this study, aiming to categorize oxacillin-resistant CoNS species recovered from blood culture specimens of inpatients at the UNESP Hospital das Clínicas in Botucatu, Brazil, over a 20-year period, and determine their sensitivity to other antimicrobial agents. The mecA gene was detected in 222 (74%) CoNS samples, and the four types of staphylococcal chromosomal cassette mec (SCCmec) were characterized in 19.4%, 3.6%, 54.5%, and 14.4% of specimens, respectively, for types I, II, III, and IV. Minimal inhibitory concentration (MIC) values to inhibit 50% (MIC50) and 90% (MIC90) of specimens were, respectively, 2 and >256µL/mL for oxacillin, 1.5 and 2µL/mL for vancomycin, 0.25 and 0.5µL/mL for linezolid, 0.094 and 0.19µL/mL for daptomycin, 0.19 and 0.5µL/mL for quinupristin/dalfopristin, and 0.125 and 0.38µL/mL for tigecycline. Resistance to oxacillin and tigecycline and intermediate resistance to quinupristin/dalfopristin were observed. Eight (2.7%) of all 300 CoNS specimens studied showed reduced susceptibility to vancomycin. Results from this study show high resistance rates of CoNS to antimicrobial agents, reflecting the necessity of using these drugs judiciously and controlling nosocomial dissemination of these pathogens.


Subject(s)
Anti-Bacterial Agents/pharmacology , Coagulase/metabolism , Drug Resistance, Bacterial/genetics , Staphylococcal Infections/microbiology , Staphylococcus/drug effects , Bacterial Proteins/genetics , Drug Resistance, Bacterial/drug effects , Genes, Bacterial/genetics , Hospitals, Teaching , Humans , Microbial Sensitivity Tests , Penicillin-Binding Proteins/genetics , Polymerase Chain Reaction , Staphylococcus/chemistry , Staphylococcus/genetics
9.
Front Microbiol ; 10: 40, 2019.
Article in English | MEDLINE | ID: mdl-30745897

ABSTRACT

Bacterial biofilms play an important role in urinary tract infections (UTIs), being responsible for persistent infections that lead to recurrences and relapses. Staphylococcus saprophyticus is one of the main etiological agents of UTIs, however, little is known about biofilm production in this species and especially about its response to the antimicrobial agents used to treat UTIs when a biofilm is present. For this reason, the aim of this work was to evaluate the response of S. saprophyticus biofilms to five antimicrobial agents. Staphylococcus saprophyticus was evaluated for antimicrobial susceptibility in its planktonic form by means of minimum inhibitory concentration (MIC) and in biofilms by means of minimum inhibitory concentration in biofilm (MICB) against the following antimicrobial agents by the microdilution technique: vancomycin, oxacillin, trimethoprim/sulfamethoxazole, ciprofloxacin, and norfloxacin. Of the 169 S. saprophyticus studied, 119 produced a biofilm as demonstrated by the polystyrene plate adherence method. Biofilm cells of S. saprophyticus exhibited a considerable increase in MICB when compared to the planktonic forms, with an increase of more than 32 times in the MICB of some drugs. Some isolates switched from the category of susceptible in the planktonic condition to resistant in the biofilm state. Statistical analysis of the results showed a significant increase in MICB (p < 0.0001) for all five drugs tested in the biofilm state compared to the planktonic form. Regarding determination of the minimum bactericidal concentration in biofilm (MBCB), there were isolates for which the minimum bactericidal concentration of all drugs was equal to or higher than the highest concentration tested.

10.
Trop Med Int Health ; 24(3): 339-347, 2019 03.
Article in English | MEDLINE | ID: mdl-30549385

ABSTRACT

OBJECTIVE: To evaluate the molecular epidemiology and to georeference Staphylococcus aureus isolated from wounds and nares of patients seen at Basic Health Units (BHUs) of a Brazilian city. METHODS: Observational, cross-sectional study conducted from 2010 to 2013. A total of 119 S. aureus strains isolated from the wounds and nares of 88 patients were studied. The isolates were characterised by identifying virulence genes encoding enterotoxins A-E, haemolysins α, ß and δ, exfoliatins A, B and D, biofilm production, Panton-Valentine Leukocidin and toxic shock syndrome toxin 1, and by pulsed-field gel electrophoresis (PFGE), multilocus sequence and spa typing. RESULTS: Eighteen methicillin-resistant Staphylococcus aureus (MRSA) (6 SCCmec type II and 12 SCCmec type IV) and 101 (85%) MSSA were identified. PFGE typing resulted in the formation of eight clusters, with STs 1, 5, 8, 30, 188, 1176 and 1635 and spa type t002 being the predominant types among MSSA. The 18 MRSA belonged to STs 5, 8 and 1176 and spa types t002 and t062. CONCLUSION: The results demonstrate widespread dissemination of MSSA and MRSA clones carrying haemolysin, biofilm and toxin genes. Kernel density estimation revealed the highest density of S. aureus in the 4, 5 and 8 BHUs.


OBJECTIF: Evaluer l'épidémiologie moléculaire et géoréférencer le Staphylococcus aureus isolé de plaies et de narines de patients vus dans les unités sanitaires de base (BHU) d'une ville brésilienne. MÉTHODES: Etude observationnelle transversale réalisée de 2010 à 2013. Au total, 119 souches de S. aureus isolées de plaies et de narines de 88 patients ont été étudiées. Les isolats ont été caractérisés par l'identification de gènes de virulence codant pour les entérotoxines AE, les hémolysines α, ß et δ, les exfoliatines A, B et D, la production de biofilm, la leucocidine de Panton-Valentine et la toxine 1 du syndrome de choc toxique, et par typage par électrophorèse sur gel en champ pulsé (PFGE), séquence multilocus et spa. RÉSULTATS: Dix-huit SARM (6 de type II SCCmec et 12 de type IV SCCmec) et 101 (85%) SASM ont été identifiés. Le typage PFGE a résulté à l'obtention de huit grappes, dont STs 1, 5, 8, 30, 188, 1176 et 1635 et le type spa t002 étant les types prédominants parmi les SASM. Les 18 SARM appartenaient aux STs 5, 8 et 1176 et aux types de spa t002 et t062. CONCLUSION: Les résultats démontrent une dissémination étendue des clones de SASM et de SARM portant les gènes de l'hémolysine, de biofilm et de toxine. L'estimation de la densité par noyau a révélé la densité la plus élevée de S. aureus dans les 4, 5 et 8 BHU.


Subject(s)
Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/epidemiology , Wounds and Injuries/microbiology , Aged , Brazil/epidemiology , Cross-Sectional Studies , Female , Humans , Male , Middle Aged , Molecular Epidemiology , Staphylococcal Infections/microbiology
11.
Braz J Infect Dis ; 22(2): 99-105, 2018.
Article in English | MEDLINE | ID: mdl-29548778

ABSTRACT

INTRODUCTION: Staphylococcus spp. - both S. aureus, including methicillin-resistant strains (MRSA) and coagulase negative staphylococci (CoNS) - are relevant agents of healthcare-associated infections. Therefore, the rapid recognition of MRSA and methicillin-resistant CoNS from blood stream infections is critically important for patient management. It is worth noting that inappropriate empiric therapy has been associated with higher in-hospital mortality. MATERIAL AND METHODS: In this study we evaluated a multiplex polymerase chain reaction (multiplex PCR) standardized to detect Staphylococcus spp., S. aureus, and mecA gene-encoded oxacillin resistance directly from blood culture bottles. A total of 371 blood cultures with Gram-positive microorganisms confirmed by Gram-stain were analyzed. Results from multiplex PCR were compared to phenotypic characterization of isolates. RESULTS: Staphylococcus aureus was detected in 85 (23.0%) blood cultures and CoNS in 286 (77.0%). There was 100% agreement between phenotypic and multiplex PCR identification. Forty-three (50.6%) of the 85 S. aureus carried the mecA gene and among the 286 CoNS, 225 (78.7%) were positive for the mecA gene. CONCLUSIONS: The multiplex PCR assay developed here was found to be sensitive, specific, rapid, and showed good agreement with the phenotypic results besides being less expensive. This PCR method could be used in clinical laboratories for rapid identification and initiation of specific and effective treatment, reducing patient mortality and morbidity. Furthermore, this method may reduce misuse of antimicrobial classes that are more expensive and toxic, thus contributing to the selection of antibiotic-resistant Staphylococcus spp.


Subject(s)
Bacteremia/diagnosis , Bacterial Proteins/genetics , Blood/microbiology , Methicillin-Resistant Staphylococcus aureus/genetics , Multiplex Polymerase Chain Reaction , Penicillin-Binding Proteins/genetics , Staphylococcal Infections/diagnosis , Anti-Bacterial Agents/pharmacology , Bacteremia/microbiology , Bacterial Proteins/isolation & purification , Blood Culture , DNA, Bacterial/genetics , Humans , Oxacillin/pharmacology , Penicillin-Binding Proteins/isolation & purification , Staphylococcus aureus/drug effects , Staphylococcus aureus/isolation & purification
12.
Braz. j. infect. dis ; Braz. j. infect. dis;22(2): 99-105, Mar.-Apr. 2018. tab, graf
Article in English | LILACS | ID: biblio-951629

ABSTRACT

ABSTRACT Introduction: Staphylococcus spp. - both S. aureus, including methicillin-resistant strains (MRSA) and coagulase negative staphylococci (CoNS) - are relevant agents of healthcare-associated infections. Therefore, the rapid recognition of MRSA and methicillin-resistant CoNS from blood stream infections is critically important for patient management. It is worth noting that inappropriate empiric therapy has been associated with higher in-hospital mortality. Material and methods: In this study we evaluated a multiplex polymerase chain reaction (multiplex PCR) standardized to detect Staphylococcus spp., S. aureus, and mecA gene-encoded oxacillin resistance directly from blood culture bottles. A total of 371 blood cultures with Gram-positive microorganisms confirmed by Gram-stain were analyzed. Results from multiplex PCR were compared to phenotypic characterization of isolates. Results: Staphylococcus aureus was detected in 85 (23.0%) blood cultures and CoNS in 286 (77.0%). There was 100% agreement between phenotypic and multiplex PCR identification. Forty-three (50.6%) of the 85 S. aureus carried the mecA gene and among the 286 CoNS, 225 (78.7%) were positive for the mecA gene. Conclusions: The multiplex PCR assay developed here was found to be sensitive, specific, rapid, and showed good agreement with the phenotypic results besides being less expensive. This PCR method could be used in clinical laboratories for rapid identification and initiation of specific and effective treatment, reducing patient mortality and morbidity. Furthermore, this method may reduce misuse of antimicrobial classes that are more expensive and toxic, thus contributing to the selection of antibiotic-resistant Staphylococcus spp.


Subject(s)
Humans , Bacterial Proteins/genetics , Blood/microbiology , Bacteremia/diagnosis , Penicillin-Binding Proteins/genetics , Methicillin-Resistant Staphylococcus aureus/genetics , Multiplex Polymerase Chain Reaction , Oxacillin/pharmacology , Staphylococcal Infections/diagnosis , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/drug effects , Bacterial Proteins/isolation & purification , DNA, Bacterial/genetics , Bacteremia/microbiology , Penicillin-Binding Proteins/isolation & purification , Blood Culture , Anti-Bacterial Agents/pharmacology
13.
Braz. j. microbiol ; Braz. j. microbiol;48(1): 159-166, Jan.-Mar. 2017. tab, graf
Article in English | LILACS | ID: biblio-839333

ABSTRACT

Abstract Staphylococcus aureus and Staphylococcus saprophyticus are the most common and most important staphylococcal species associated with urinary tract infections. The objective of the present study was to compare and to evaluate the accuracy of four phenotypic methods for the detection of beta-lactamase production in Staphylococcus spp. Seventy-three strains produced a halo with a diameter ≤28 mm (penicillin resistant) and all of them were positive for the blaZ gene. Among the 28 susceptible strain (halo ≥29 mm), 23 carried the blaZ gene and five did not. The zone edge test was the most sensitive (90.3%), followed by MIC determination (85.5%), but the specificity of the former was low (40.0%). The nitrocefin test was the least sensitive (28.9%). However, the nitrocefin test together with the disk diffusion method showed the highest specificity (100%). The present results demonstrated that the zone edge test was the most sensitive phenotypic test for detection of beta-lactamase, although it is still not an ideal test to detect this type of resistance since its specificity was low. However, the inhibition halo diameter of the penicillin disk can be used together with the zone edge test since the same disk is employed in the two tests. Combined analysis of the two tests shows a sensitivity of 90.3% and specificity of 100%, proving better sensitivity, especially for S. saprophyticus. This is a low-cost test of easy application and interpretation that can be used in small and medium-sized laboratories where susceptibility testing is usually performed by the disk diffusion method.


Subject(s)
beta-Lactamases/genetics , beta-Lactamases/metabolism , Microbial Sensitivity Tests , beta-Lactam Resistance , Staphylococcal Infections/microbiology , Urinary Tract Infections/microbiology , Penicillin Resistance , Sensitivity and Specificity , Disk Diffusion Antimicrobial Tests , Staphylococcus saprophyticus/drug effects , Staphylococcus saprophyticus/genetics , Staphylococcus saprophyticus/metabolism , Genotype
14.
Braz. J. Microbiol. ; 48(1): 159-166, jan.-mar. 2017. graf, ilus, tab
Article in English | VETINDEX | ID: vti-22688

ABSTRACT

Staphylococcus aureus and Staphylococcus saprophyticus are the most common and most important staphylococcal species associated with urinary tract infections. The objective of the present study was to compare and to evaluate the accuracy of four phenotypic methods for the detection of beta-lactamase production in Staphylococcus spp. Seventy-three strains produced a halo with a diameter ≤ 28 mm (penicillin resistant) and all of them were positive for the blaZ gene. Among the 28 susceptible strain (halo ≥ 29 mm), 23 carried the blaZ gene and five did not. The zone edge test was the most sensitive (90.3%), followed by MIC determination (85.5%), but the specificity of the former was low (40.0%). The nitrocefin test was the least sensitive (28.9%). However, the nitrocefin test together with the disk diffusion method showed the highest specificity (100%). The present results demonstrated that the zone edge test was the most sensitive phenotypic test for detection of beta-lactamase, although it is still not an ideal test to detect this type of resistance since its specificity was low. However, the inhibition halo diameter of the penicillin disk can be used together with the zone edge test since the same disk is employed in the two tests. Combined analysis of the two tests shows a sensitivity of 90.3% and specificity of 100%, proving better sensitivity, especially for S. saprophyticus. This is a low-cost test of easy application and interpretation that can be used in small and medium-sized laboratories where susceptibility testing is usually performed by the disk diffusion method.(AU)


Subject(s)
Staphylococcus , Urinary Tract/pathology , beta-Lactamases/genetics , Phenotype
15.
Braz J Microbiol ; 48(1): 159-166, 2017.
Article in English | MEDLINE | ID: mdl-27889420

ABSTRACT

Staphylococcus aureus and Staphylococcus saprophyticus are the most common and most important staphylococcal species associated with urinary tract infections. The objective of the present study was to compare and to evaluate the accuracy of four phenotypic methods for the detection of beta-lactamase production in Staphylococcus spp. Seventy-three strains produced a halo with a diameter ≤28mm (penicillin resistant) and all of them were positive for the blaZ gene. Among the 28 susceptible strain (halo ≥29mm), 23 carried the blaZ gene and five did not. The zone edge test was the most sensitive (90.3%), followed by MIC determination (85.5%), but the specificity of the former was low (40.0%). The nitrocefin test was the least sensitive (28.9%). However, the nitrocefin test together with the disk diffusion method showed the highest specificity (100%). The present results demonstrated that the zone edge test was the most sensitive phenotypic test for detection of beta-lactamase, although it is still not an ideal test to detect this type of resistance since its specificity was low. However, the inhibition halo diameter of the penicillin disk can be used together with the zone edge test since the same disk is employed in the two tests. Combined analysis of the two tests shows a sensitivity of 90.3% and specificity of 100%, proving better sensitivity, especially for S. saprophyticus. This is a low-cost test of easy application and interpretation that can be used in small and medium-sized laboratories where susceptibility testing is usually performed by the disk diffusion method.


Subject(s)
Microbial Sensitivity Tests , beta-Lactam Resistance , beta-Lactamases/genetics , beta-Lactamases/metabolism , Disk Diffusion Antimicrobial Tests , Genotype , Penicillin Resistance , Sensitivity and Specificity , Staphylococcal Infections/microbiology , Staphylococcus saprophyticus/drug effects , Staphylococcus saprophyticus/genetics , Staphylococcus saprophyticus/metabolism , Urinary Tract Infections/microbiology
16.
Braz J Microbiol ; 46(2): 535-43, 2015 Jun.
Article in English | MEDLINE | ID: mdl-26273271

ABSTRACT

The objective of this study was to characterize the clonal profile, virulence factors and antimicrobial resistance, particularly oxacillin resistance, of Staphylococcus aureus isolated from sheep milk. Milk samples were collected from all teats for the California Mastitis Test (CMT), somatic cell count, identification of S. aureus, investigation in these strains of genes encoding toxins (sea, seb, sec, sed, tst), biofilm (icaA, icaC, icaD, bap), leukocidin (luk-PV) oxacillin resistance by mecA gene detection and susceptibility testing (12 antibiotics). Messenger RNA expression was evaluated by RT-PCR in isolates carrying toxin and biofilm genes. Biofilm formation was also evaluated phenotypically by adherence to polystyrene plates. The clonal profile of S. aureus was investigated by pulsed-field gel electrophoresis. A total of 473 milk samples were collected from 242 animals on three farms and 20 S. aureus strains were isolated and none carried the mecA gene. The two sec gene-positive isolates and the isolates carrying the tst and luk-PV genes were positive by RT-PCR. Staphylococcus aureus isolated from the three flocks studied showed high susceptibility to the drugs tested and none was biofilm producer, indicating that biofilm formation was not a virulence factor causing infection by these strains. The typing of 17 S. aureus isolates revealed the presence of a common clone on the three farms studied, and the presence and expression of the sec and tst genes in one strain of this clone suggest the possible acquisition of virulence genes by this clone, a fact that is important for animal health and food hygiene.


Subject(s)
Milk/microbiology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/physiology , Animals , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , California , Electrophoresis, Gel, Pulsed-Field , Gene Expression Profiling , Microbial Sensitivity Tests , Molecular Typing , Reverse Transcriptase Polymerase Chain Reaction , Sheep , Staphylococcus aureus/classification , Staphylococcus aureus/drug effects , Virulence Factors/genetics
17.
Braz. j. microbiol ; Braz. j. microbiol;46(2): 535-543, Apr-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-749725

ABSTRACT

The objective of this study was to characterize the clonal profile, virulence factors and antimicrobial resistance, particularly oxacillin resistance, of Staphylococcus aureus isolated from sheep milk. Milk samples were collected from all teats for the California Mastitis Test (CMT), somatic cell count, identification of S. aureus, investigation in these strains of genes encoding toxins (sea, seb, sec, sed, tst), biofilm (icaA, icaC, icaD, bap), leukocidin (luk-PV) oxacillin resistance by mecA gene detection and susceptibility testing (12 antibiotics). Messenger RNA expression was evaluated by RT-PCR in isolates carrying toxin and biofilm genes. Biofilm formation was also evaluated phenotypically by adherence to polystyrene plates. The clonal profile of S. aureus was investigated by pulsed-field gel electrophoresis. A total of 473 milk samples were collected from 242 animals on three farms and 20 S. aureus strains were isolated and none carried the mecA gene. The two sec gene-positive isolates and the isolates carrying the tst and luk-PV genes were positive by RT-PCR. Staphylococcus aureus isolated from the three flocks studied showed high susceptibility to the drugs tested and none was biofilm producer, indicating that biofilm formation was not a virulence factor causing infection by these strains. The typing of 17 S. aureus isolates revealed the presence of a common clone on the three farms studied, and the presence and expression of the sec and tst genes in one strain of this clone suggest the possible acquisition of virulence genes by this clone, a fact that is important for animal health and food hygiene.


Subject(s)
Animals , Milk/microbiology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/physiology , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , California , Electrophoresis, Gel, Pulsed-Field , Gene Expression Profiling , Microbial Sensitivity Tests , Molecular Typing , Reverse Transcriptase Polymerase Chain Reaction , Sheep , Staphylococcus aureus/classification , Staphylococcus aureus/drug effects , Virulence Factors/genetics
18.
Braz. J. Microbiol. ; 46(2): 535-543, Apr.-Jun. 2015. tab, graf, ilus
Article in English | VETINDEX | ID: vti-481380

ABSTRACT

The objective of this study was to characterize the clonal profile, virulence factors and antimicrobial resistance, particularly oxacillin resistance, of Staphylococcus aureus isolated from sheep milk. Milk samples were collected from all teats for the California Mastitis Test (CMT), somatic cell count, identification of S. aureus, investigation in these strains of genes encoding toxins (sea, seb, sec, sed, tst), biofilm (icaA, icaC, icaD, bap), leukocidin (luk-PV) oxacillin resistance by mecA gene detection and susceptibility testing (12 antibiotics). Messenger RNA expression was evaluated by RT-PCR in isolates carrying toxin and biofilm genes. Biofilm formation was also evaluated phenotypically by adherence to polystyrene plates. The clonal profile of S. aureus was investigated by pulsed-field gel electrophoresis. A total of 473 milk samples were collected from 242 animals on three farms and 20 S. aureus strains were isolated and none carried the mecA gene. The two sec gene-positive isolates and the isolates carrying the tst and luk-PV genes were positive by RT-PCR. Staphylococcus aureus isolated from the three flocks studied showed high susceptibility to the drugs tested and none was biofilm producer, indicating that biofilm formation was not a virulence factor causing infection by these strains. The typing of 17 S. aureus isolates revealed the presence of a common clone on the three farms studied, and the presence and expression of the sec and tst genes in one strain of this clone suggest the possible acquisition of virulence genes by this clone, a fact that is important for animal health and food hygiene.(AU)


Subject(s)
Animals , Milk/microbiology , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/physiology , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , California , Electrophoresis, Gel, Pulsed-Field , Gene Expression Profiling , Microbial Sensitivity Tests , Molecular Typing , Reverse Transcriptase Polymerase Chain Reaction , Sheep , Staphylococcus aureus/classification , Staphylococcus aureus , Virulence Factors/genetics
19.
Ci. Anim. bras. ; 16(1): 116-124, Jan-Mar. 2015. tab
Article in Portuguese | VETINDEX | ID: vti-9653

ABSTRACT

The aim of this study was to evaluate the occurrence of ovine subclinical mastitis in two lactations, and the disease evolution in two consecutive lactations. Milk samples originated from a herd with 160 Santa Inês breed sheep. In the first lactation, milk samples were collected from ewes at 14 days post-partum and during the period near weaning. In the second lactation, milk samples were collected from the same animals at 14 days postpartum, at 52 days postpartum and at weaning. Screening of subclinical mastitis was carried out preliminarily by means of California Mastitis Test (CMT). Milk samples for microbiological culture were obtained from CMT-positive and CMT-negative mammary halves. Coagulase-negative Staphylococci were the most prevalent microorganisms isolated in two lactations. Ewes that remained with subclinical mastitis in lactation 1 were predominant. Spontaneous recoveries were significantly lower in the period between 52 days of lactation and weaning of lambs. No difference was observed regarding the progression of subclinical mastitis cases compared the two lactations (P>0.05).(AU)


O objetivo desse estudo foi avaliar a ocorrência da mastite subclínica ovina em duas lactações e a evolução da doença em duas lactações consecutivas. As amostras de leite foram originadas de um rebanho com 160 ovelhas da raça Santa Inês. Na primeira lactação, colheram-se amostras em ovelhas aos 14 dias pós-parto e no período próximo ao desmame. Na segunda lactação, as amostras de leite foram colhidas dos mesmos animais aos 14 dias pós-parto, 52 dias pós-parto e ao desmame. A triagem dos casos de mastite subclínica foi feita preliminarmente por meio do California Mastitis Test (CMT). As amostras de leite para a investigação microbiológica foram obtidas de metades mamárias positivas e negativas ao CMT. Os estafilococos coagulase-negativos foram os micro-organismos de maior ocorrência nas duas lactações. Houve a predominância de ovelhas que permaneceram com mastite subclínica durante a Lactação 1. As curas espontâneas foram  significativamente inferiores no período entre 52 dias de lactação e o desmame dos cordeiros. Nenhuma diferença foi observada quanto à evolução dos casos de mastite subclínica infecciosa quando comparadas duas lactações (P>0,05).(AU)


Subject(s)
Animals , Sheep Diseases , Sheep/anatomy & histology , Mastitis/veterinary , Lactation , Milk/physiology , Microbiological Techniques/methods
20.
Ciênc. anim. bras. (Impr.) ; 16(1): 116-124, Jan-Mar. 2015. tab
Article in Portuguese | VETINDEX | ID: biblio-1473373

ABSTRACT

The aim of this study was to evaluate the occurrence of ovine subclinical mastitis in two lactations, and the disease evolution in two consecutive lactations. Milk samples originated from a herd with 160 Santa Inês breed sheep. In the first lactation, milk samples were collected from ewes at 14 days post-partum and during the period near weaning. In the second lactation, milk samples were collected from the same animals at 14 days postpartum, at 52 days postpartum and at weaning. Screening of subclinical mastitis was carried out preliminarily by means of California Mastitis Test (CMT). Milk samples for microbiological culture were obtained from CMT-positive and CMT-negative mammary halves. Coagulase-negative Staphylococci were the most prevalent microorganisms isolated in two lactations. Ewes that remained with subclinical mastitis in lactation 1 were predominant. Spontaneous recoveries were significantly lower in the period between 52 days of lactation and weaning of lambs. No difference was observed regarding the progression of subclinical mastitis cases compared the two lactations (P>0.05).


O objetivo desse estudo foi avaliar a ocorrência da mastite subclínica ovina em duas lactações e a evolução da doença em duas lactações consecutivas. As amostras de leite foram originadas de um rebanho com 160 ovelhas da raça Santa Inês. Na primeira lactação, colheram-se amostras em ovelhas aos 14 dias pós-parto e no período próximo ao desmame. Na segunda lactação, as amostras de leite foram colhidas dos mesmos animais aos 14 dias pós-parto, 52 dias pós-parto e ao desmame. A triagem dos casos de mastite subclínica foi feita preliminarmente por meio do California Mastitis Test (CMT). As amostras de leite para a investigação microbiológica foram obtidas de metades mamárias positivas e negativas ao CMT. Os estafilococos coagulase-negativos foram os micro-organismos de maior ocorrência nas duas lactações. Houve a predominância de ovelhas que permaneceram com mastite subclínica durante a Lactação 1. As curas espontâneas foram  significativamente inferiores no período entre 52 dias de lactação e o desmame dos cordeiros. Nenhuma diferença foi observada quanto à evolução dos casos de mastite subclínica infecciosa quando comparadas duas lactações (P>0,05).


Subject(s)
Animals , Sheep Diseases , Lactation , Mastitis/veterinary , Sheep/anatomy & histology , Milk/physiology , Microbiological Techniques/methods
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