Coagulase-positive staphylococci (CPS) cause staphylococcal food poisoning. Recently, these bacteria have received increasing attention due to their potential role in the dissemination of antibiotic resistance markers. The present study aimed to evaluate coagulase-positive staphylococci counts, species distribution, enterotoxin genes prevalence, and the antibiotic resistance profile of CPS isolated from in natura chicken meat. Fifteen frozen and 15 chilled industrialized, uncooked chicken parts or entire carcasses were used. Staphylococcal counts revealed that frozen chicken meat samples displayed the lowest CPS count compared with chilled chicken meat samples (p<0.01). Staphylococcus aureus (62%) was the most common species, followed by S. intermedius, S. delphini, and S. schleiferi subsp. coagulans (10% each) and S. hyicus (8%). The polymerase chain reaction identification of sea, seb, sec, sed, and see genes revealed that 70% of the isolates harbored at least one enterotoxin gene, with sea and sed being the most frequently encountered ones. Two of the 50 investigated strains harbored three different enterotoxin genes. A high frequency of isolates resistant to penicillin, teicoplanin, oxacillin, and clindamycin was observed, and 80% of CPS were found to be resistant to at least one of the 11 tested antimicrobials. Vancomycin-resistant S. aureus and S. intermedius showed minimum inhibitory concentrations of 512 and 64 μg/mL, respectively. These isolates might indicate the dissemination of vancomycin resistance in the community and imply food safety hazards.
The aim of this study was to evaluate the species distribution, antibiotic-resistance profile and presence of enterotoxin (SE) genes in staphylococci isolated from the Dilúvio stream in South Brazil. Eighty-eight staphylococci were identified, 93.18% were identified as coagulase-negative (CNS) and 6.82% coagulasepositive (CPS). Fourteen Staphylococcus species were detected and the most frequently were Staphylococcus cohnii (30.48%) and S. haemolyticus (21.95%). Resistance to erythromycin was verified in 37.50% of the strains, followed by 27.27% to penicillin, 12.50% to clindamycin, 6.81% to trimethoprim-sulfamethoxazole, 5.68% to chloramphenicol and 2.27% to norfloxacin. None of the investigated strains showed gentamicin and ciprofloxacin resistance. The strains were tested for the presence of sea, seb, sec, sed and see genes by PCR and only CNS strains (43.18%) showed positive results to one or more SE genes. The scientific importance of our results is due to the lack of data about these topics in polluted waters in Brazil. In conclusion, polluted waters from the Dilúvio stream may constitute a reservoir for disseminating antibiotic-resistance and enterotoxin into the community. In addition, the detection of staphylococci in the polluted waters of the Dilúvio stream indicated a situation of environmental contamination and poor sanitation conditions.
Penicillium brasilianum was previously isolated from tea and identified by molecular biology technique. A PlackettBurman design, followed by a complete second order design was used for the screening of most important factors and to maximize the pectin liase (PMGL) activity, respectively. The maximum PMGL activity by P. brasilianum achieved was 9.0 U/mL after 48 h of cultivation in a medium containing pectin (33.0 g/L), yeast extract (30.0 g/L) and potassium phosphate (2.0 g/L) at 30ºC, with a stirring rate of 180 rpm, initial pH 5.5 and 5x106 spores/mL inoculum size. The kinetic evaluation in terms of substrate consumption demonstrated that the maximum production of PMGL was at 72 h, and 40% of the total organic carbon, 25% of the nitrogen, 88% of the magnesium, 13% of the potassium and 66% of the iron were consumed. The pH remained almost stable during the whole period of production (5.33 to 4.9). The partial characterization of the crude PMGL enzyme extract showed optimal pH and temperature of 5.5 and 37°C, respectively.
This report demonstrates the usefulness of PCR for the genes spaS and sboA as a means of identifying Bacillus strains with a potential to produce subtilin and subtilosin A. One collection strain and five Bacillus spp. isolated from aquatic environments in the Amazon basin were screened by PCR using primers for sboA and spaS designed specifically for this study. The sequences of the PCR products showed elevated homology with previously described spaS and sboA genes. Antimicrobial peptides were isolated from culture supernatants and analyzed by mass spectrometry. For all samples, the mass spectra revealed clusters with peaks at m/z 3300–3500 Da, corresponding to subtilosin A, subtilin and isoforms of these peptides. These results suggest that the antimicrobial activity of these strains may be associated with the production of subtilosin A and/or subtilin. The PCR used here was efficient in identifying novel Bacillus strains with the essential genes for producing subtilosin A and subtilin.
The pink fairy armadillo Chlamyphorus truncatus is the smallest extant armadillo and one of the least-known fossorial mammals. The aim of this study was to establish if its middle ear is specially adapted to the subterranean environment, through comparison with more epigeic relatives of the groups Euphractinae (Chaetophractus villosus, Chaetophractus vellerosus, Zaedyus pichiy) and Dasypodinae (Dasypus hybridus). We examined the middle ears using micro-computed tomography and subsequent three-dimensional reconstructions. D. hybridus has a relatively small middle ear cavity, an incomplete bulla and 'ancestral' ossicular morphology. The other species, including Chlamyphorus, have fully ossified bullae and middle ear ossicles, with a morphology between 'transitional' and 'freely mobile', but in all armadillos the malleus retains a long anterior process. Unusual features of armadillo ears include the lack of a pedicellate lenticular apophysis and the presence, in some species, of an element of Paaw within the stapedius muscle. In common with many subterranean mammals, Chlamyphorus has a relatively flattened malleo-incudal articulation and appears to lack a functional tensor tympani muscle. Its middle ear cavity is not unusually enlarged, and its middle ear ossicles seem less robust than those of the other armadillos studied. In comparison with the euphractines, there is no reason to believe that the middle ear of this species is specially adapted to the subterranean environment; some aspects may even be indicative of degeneration. The screaming hairy armadillo, Chaetophractus vellerosus, has the most voluminous middle ear in both relative and absolute terms. Its hypertrophied middle ear cavity likely represents an adaptation to low-frequency hearing in arid rather than subterranean conditions.
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