Molecular characterization and clonal diversity of methicillin-susceptible Staphylococcus aureus in milk of cows with mastitis in Brazil

Mastitis is an important disease for the dairy industry worldwide, causing economic losses and reducing milk quality and production. Staphylococcus aureus is a worldwide agent of this intramammary infection, which also causes foodborne diseases. The objective of this study was to determine the frequency of methicillin-susceptible Staphylococcus aureus (MSSA) isolates in milk of mastitis cows in Brazil and to analyze the genetic lineages and the content of antimicrobial resistance genes and virulence factors among these isolates. Fifty-six MSSA isolates were recovered from 1,484 milk samples (positive for the California mastitis test) of 518 cows from 11 different farms in Brazil (representing 51% of total Staph. aureus obtained), and they were further characterized. Methicillin-susceptible Staphylococcus aureus were isolated from 3.7% of California mastitis test-positive tested milk samples and from 6.2% of tested mastitic cows. Methicillin-susceptible Staphylococcus aureus isolates were characterized by spa typing, agr typing, and multilocus sequence typing, and resistance and virulence traits were investigated by PCR. Seven spa types were identified among MSSA (% of isolates): t127 (44.6), t605 (37.5), t002, t1784, t2066 (1.8), and 2 new ones: t10856 (10.7) and t10852 (1.8). Five distinct sequence types (ST) were detected (% of isolates): ST1 (46.4), ST126 (37.5), ST133 (10.7), ST5 (3.6), and a novel ST registered as ST2493 (1.8). Resistances were detected for streptomycin, chloramphenicol, and tetracycline. One strain contained the chloramphenicol resistance gene (fexA; included within transposon Tn558) and 3 strains contained the tetracycline resistance gene [tet(K)]. Methicillin-susceptible Staphylococcus aureus strains were susceptible to most of the antibiotics studied and lacked the virulence genes of Panton-Valentine leukocidin (lukF/S-PV), toxic shock syndrome toxin 1 (tst), exfoliative toxin A (eta), and exfoliative toxin B (etb), as well as the genes of the immune evasion cluster. Methicillin-susceptible Staphylococcus aureus isolates were detected in a relatively low proportion of cows with mastitis (6.2%) and recovered isolates presented high diversity of genetic lineages, with CC1 and CC126 the predominant clonal complexes, and CC133 also being detected. Larger epidemiological studies with molecular characterization of isolates are required to deepen the knowledge on the circulating genetic lineages among the cow population with mastitis. © 2013 American Dairy Science Association.

Influence of the resistance training on heart rate variability, functional capacity and muscle strength in the chronic obstructive pulmonary disease

Background. The chronic obstructive pulmonary disease (COPD) is associated with the strength and resistance decreasing in addition to the dysfunction on autonomic nervous system (ANS). The aerobic training isolated or in association with the resistance training showed evidence of beneficial effects on an autonomic modulation of COPD; however, there are no studies addressing the effect of isolated resistance training.Aims. This study aims at investigating the influence of resistance training on an autonomic modulation through heart rate variability (HRV), functional capacity and muscle strength in individuals with COPD.Design. Clinical series study.Setting. Outpatients.Population. The study involved 13 individuals with COPD.Methods. The experimental protocol was composed by an initial and final evaluation that consisted in autonomic evaluations (HRV), cardiopulmonary functional capacity evaluation (6-minute walk test) and strength evaluation (dynamometry) in addition by the resistance training performed by 24 sessions lasted 60 minutes each one and on a frequency of three times a week. The intensity was determined initially with 60% of one maximum repetition and was progressively increased in each five sessions until 80%.Results. The HRV temporal and spectral indexes analysis demonstrates improvement of autonomic modulation, with significant statistical increases to sympathetic and parasympathetic components of ANS representing by SDNN, LF and HF. In addition, it was observed significant statistical increases to shoulder abduction and. knee flexion strength and functional capacity.Conclusion. The exclusive resistance training performed was able to positively influence the autonomic modulation; in addition it promoted benefits on cardiorespiratory functional capacity and strength benefits in individuals with COPD.Clinical Rehabilitation Impact. This study could contribute to clinical and professionals researchers that act with COPD, even though the resistance component of pulmonary rehabilitation presents consensual benefits on several healthy indicators parameters. There is no evidence about the effects on HRV before. Moreover, this study showed, on clinical practice, the HRV uses as an ANS activity on sinus node evaluation and highlights further importance on scientific context.

Anti-Trichophyton Activity of Protocatechuates and Their Synergism with Fluconazole

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Glycolipid Sensing and Innate Immunity in Paracoccidioidomycosis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Densities and antimicrobial resistance of Escherichia coli isolated from marine waters and beach sands

Bacterial resistance is a rising problem all over the world. Many studies have showed that beach sands can contain higher concentration of microorganisms and represent a risk to public health. This paper aims to evaluate the densities and resistance to antimicrobials of Escherichia coli strains, isolated from seawater and samples. The hypothesis is that microorganisms show higher densities in contaminated beach sands and more antimicrobial resistance than the water column. Density, distribution, and antimicrobial resistance of bacteria E. coli were evaluate in seawater and sands from two recreational beaches with different levels of pollution. At the beach with higher degree of pollution (Gonzaguinha), water samples presented the highest densities of E. coli; however, higher frequency of resistant strains was observe in wet sand (71.9 %). Resistance to a larger number of antimicrobial groups was observe in water (betalactamics, aminoglycosides, macrolides, rifampicins, and tetracyclines) and sand (betagalactamics and aminoglycosids). In water samples, highest frequencies of resistance were obtain against ampicilin (22.5 %), streptomycin (15.0 %), and rifampicin (15.0 %), while in sand, the highest frequencies were observe in relation to ampicilin (36.25 %) and streptomycin (23.52 %). At the less polluted beach, Ilha Porchat, highest densities of E. coli and higher frequency of resistance were obtain in wet and dry sand (53.7 and 53.8 %, respectively) compared to water (50 %). Antimicrobial resistance in strains isolated from water and sand only occurred against betalactamics (ampicilin and amoxicilin plus clavulanic acid). The frequency and variability of bacterial resistance to antimicrobials in marine recreational waters and sands were related to the degree of fecal contamination in this environment. These results show that water and sands from beaches with a high index of fecal contamination of human origin may be potential sources of contamination by pathogens and contribute to the dissemination of bacterial resistance.

Susceptibility of helminth species from horses against different chemical compounds in Brazil

By means of parasitological necropsies, the present study aimed to evaluate, in six experiments, the degree of susceptibility or resistance of different helminth species which naturally infect horses to ivermectin 0.2mg/kg, abamectin 0.2mg/kg, moxidectin 0.4mg/kg, trichlorfon 35mg/kg, ivermectin 0.2mg/kg+praziquantel 2.5mg/kg, abamectin 0.2mg/kg+praziquantel 2.5mg/kg and ivermectin 0.2mg/kg+6.6 mg/kg pyrantel. At experimental day zero, the horses were allocated to treatment groups based on average counts of strongylid eggs per gram of feces (EPG) obtained on days -3, -2 and -1. Oxyuris sp. infections were confirmed as positive or negative. All the animals in the six experiments were naturally infected by this helminth species. Each group (control or treated) consisted of six animals. All the assessed Habronema muscae populations analyzed were susceptible to ivermectin, abamectin and moxidectin. Of the six Trichostrongylus axei populations, four were susceptible to ivermectin, abamectin, moxidectin, trichlorfon and ivermectin+praziquantel, and two were resistant to abamectin+praziquantel and ivermectin+pyrantel. Both Strongyloides westeri populations analyzed were susceptible to ivermectin, abamectin, moxidectin and abamectin+praziquantel. For O. equi, resistance was found in four different populations treated with ivermectin, abamectin, moxidectin, trichlorfon and ivermectin+praziquantel. Only combinations of abamectin+praziquantel and ivermectin+pyrantel were effective against this parasite species. All the large strongyles diagnosed in the present study (Strongyus edentatus, Strongyus vulgaris and Triodontophorus serratus) were susceptible to all the chemicals tested, with the exception of trichlorfon. Of the Cyathostominae populations, one was diagnosed as resistant to ivermectin and another to trichlorfon. The remaining populations from this nematode group were considered to be sensitive to ivermectin, abamectin, moxidectin, ivermectin+praziquantel, abamectin+praziquantel and ivermectin+pyrantel. New studies should be performed in different regions to evaluate the efficacy of trichlorfon in others field populations of helminthes.

Isolation and molecular identification of Fusobacterium nucleatum from Nigerian patients with oro-facial infections.

Fusobacterium nucleatum is one of the most common anaerobic bacteria present in the oral cavity and is often isolated from infections involving other body sites. To characterise F. nucleatum strains from patients attending a teaching hospital in Nigeria in order to provide information on the methods for accurate identification of anaerobes in clinical specimen. Fusobacterium nucleatum specie from 50 patients presenting with oro-facial infections were studied by culture on Fusobacterium selective agar and fastidious anaerobe agar. The isolates were characterised based on colonial morphology, microscopy, lipase production, susceptibility to kanamycin and colistin and resistance to vancomycin. Biochemical tests were performed using a commercial test kit. The identity of the isolates was confirmed based on molecular characterization performed using polymerase chain reaction (PCR) analysis. Forty-eight (96%) F. nucleatum isolates were obtained from the 50 patients by culture and all the isolates were identified by colonial appearance and microscopy based on their unique spindle shape with tapered ends. Only 26 (54.2%) of the 48 isolates were identified by commercial API 20A test kit while PCR confirmed the identity of all the isolates. Anaerobes are involved in human infections and their study is quite cumbersome due to tedious nature and high cost of the techniques involved. Cultural method is reliable in the isolation and identification of F. nucleatum species. PCR is a rapid and simple method that can complement the phenotypic identification of anaerobes and would assist in their full identification.

Subcritical crack growth and in vitro lifetime prediction of resin composites with different filler distributions

Objectives. Verify the influence of different filler distributions on the subcritical crack growth (SCG) susceptibility, Weibull parameters (m and sigma(0)) and longevity estimated by the strength-probability-time (SPT) diagram of experimental resin composites. Methods. Four composites were prepared, each one containing 59 vol% of glass powder with different filler sizes (d(50) = 0.5; 0.9; 1.2 and 1.9 mu m) and distributions. Granulometric analyses of glass powders were done by a laser diffraction particle size analyzer (Sald-7001, Shimadzu, USA). SCG parameters (n and sigma(f0)) were determined by dynamic fatigue (10(-2) to 10(2) MPa/s) using a biaxial flexural device (12 x 1.2 mm; n = 10). Twenty extra specimens of each composite were tested at 10(0) MPa/s to determine m and sigma(0). Specimens were stored in water at 37 degrees C for 24 h. Fracture surfaces were analyzed under SEM. Results. In general, the composites with broader filler distribution (C0.5 and C1.9) presented better results in terms of SCG susceptibility and longevity. C0.5 and C1.9 presented higher n values (respectively, 31.2 +/- 6.2(a) and 34.7 +/- 7.4(a)). C1.2 (166.42 +/- 0.01(a)) showed the highest and C0.5 (158.40 +/- 0.02(d)) the lowest sigma(f0) value (in MPa). Weibull parameters did not vary significantly (m: 6.6 to 10.6 and sigma(0): 170.6 to 176.4 MPa). Predicted reductions in failure stress (P-f = 5%) for a lifetime of 10 years were approximately 45% for C0.5 and C1.9 and 65% for C0.9 and C1.2. Crack propagation occurred through the polymeric matrix around the fillers and all the fracture surfaces showed brittle fracture features. Significance. Composites with broader granulometric distribution showed higher resistance to SCG and, consequently, higher longevity in vitro. (C) 2012 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Fracture susceptibility of endodontically treated teeth

Aim: To assess the influence of cervical preparation on fracture susceptibility of roots. Material and methods: During root canal instrumentation, the cervical portions were prepared with different taper instruments: I: no cervical preparation; II: #30/.08; III: #30/.10; IV: #70/.12. The specimens were sealed with the following filling materials (n = 8), A: unfilled; B: Endofill/gutta-percha; C: AH Plus/gutta-percha; D: Epiphany SE/Resilon. For the fracture resistance test, a universal testing machine was used at 1 mm per minute. Results: anova demonstrated difference (P < 0.05) between taper instruments with a higher value for group I (205.3 +/- 77.5 N) followed by II (185.2 +/- 70.8 N), III (164.8 +/- 48.9 N), and IV (156.7 +/- 41.4 N). There was no difference (P > 0.05) between filling materials A (189.1 +/- 66.3 N), B (186.3 +/- 61.0 N), C (159.7 +/- 69.9 N), and D (176.9 +/- 55.2 N). Conclusions: Greater cervical wear using a #70/.12 file increased the root fracture susceptibility, and the tested filling materials were not able to restore resistance.

Virulence Genes and Antimicrobial Resistance Profiles of Pasteurella multocida Strains Isolated from Rabbits in Brazil

Pasteurella multocida is responsible for a wide range of diseases in domestic animals. In rabbits, the agent is related to nasal discharge, pneumonia, otitis media, pyometra, orchitis, abscess, and septicemia. One hundred and forty rabbits with respiratory diseases from four rabbitries in Sao Paulo State, Brazil were evaluated for the detection of P. multocida in their nasal cavities. A total of twenty-nine animals were positive to P. multocida isolation, and 46 strains were selected and characterized by means of biochemical tests and PCR. P. multocida strains were tested for capsular type, virulence genes, and resistance profile. A total of 45.6% (21/46) of isolates belonged to capsular type A, and 54.34% (25/46) of the isolates were untypeable. None of the strains harboured toxA or pfhA genes. The frequency of the other twenty genes tested was variable, and the data generated was used to build a dendrogram, showing the relatedness of strains, which were clustered according to origin. Resistance revealed to be more common against sulfonamides and cotrimoxazole, followed by erythromycin, penicillin, and amoxicillin.

PRNP promoter polymorphisms are associated with BSE susceptibility in Swiss and German cattle

BACKGROUND: Non-synonymous polymorphisms within the prion protein gene (PRNP) influence the susceptibility and incubation time for transmissible spongiform encephalopathies (TSE) in some species such as sheep and humans. In cattle, none of the known polymorphisms within the PRNP coding region has a major influence on susceptibility to bovine spongiform encephalopathy (BSE). Recently, however, we demonstrated an association between susceptibility to BSE and a 23 bp insertion/deletion (indel) polymorphism and a 12 bp indel polymorphism within the putative PRNP promoter region using 43 German BSE cases and 48 German control cattle. The objective of this study was to extend this work by including a larger number of BSE cases and control cattle of German and Swiss origin. RESULTS: Allele, genotype and haplotype frequencies of the two indel polymorphisms were determined in 449 BSE cattle and 431 unaffected cattle from Switzerland and Germany including all 43 German BSE and 16 German control animals from the original study. When breeds with similar allele and genotype distributions were compared, the 23 bp indel polymorphism again showed a significant association with susceptibility to BSE. However, some additional breed-specific allele and genotype distributions were identified, mainly related to the Brown breeds. CONCLUSION: Our study corroborated earlier findings that polymorphisms in the PRNP promoter region have an influence on susceptibility to BSE. However, breed-specific differences exist that need to be accounted for when analyzing such data.

Effects of experimental inbreeding on herbivore resistance and plant fitness: the role of history of inbreeding, herbivory and abiotic factors

Inbreeding is common in plant populations and can affect plant fitness and resistance against herbivores. These effects are likely to depend on population history. In a greenhouse experiment with plants from 17 populations of Lychnis flos-cuculi, we studied the effects of experimental inbreeding on resistance and plant fitness. Depending on the levels of past herbivory and abiotic factors at the site of plant origin, we found either inbreeding or outbreeding depression in herbivore resistance. Furthermore, when not damaged experimentally by snail herbivores, plants from populations with higher heterozygosity suffered from inbreeding depression and those from populations with lower heterozygosity suffered from outbreeding depression. These effects of inbreeding and outbreeding were not apparent under experimental snail herbivory. We conclude that inbreeding effects on resistance and plant fitness depend on population history. Moreover, herbivory can mask inbreeding effects on plant fitness. Thus, understanding inbreeding effects on plant fitness requires studying multiple populations and considering population history and biotic interactions.

Multidrug- and extensively drug-resistant tuberculosis

We evaluated risk factors and treatment outcomes associated with multidrug-resistant (MDR) and extensively drug-resistant (XDR) tuberculosis (TB) in Germany in 2004–2006. In 177 (4%) of 4,557 culture-positive TB cases, Mycobacterium tuberculosis isolates were identified as MDR TB; an additional 7 (0.15%) met criteria for XDR TB. Of these 184 patients, 148 (80%) were born in countries of the former Soviet Union. In patients with XDR TB, hospitalization was longer (mean ± SD 202 ± 130 vs. 123 ± 81 days; p = 0.015) and resistance to all first-line drugs was more frequent (36% vs. 86%; p = 0.013) than in patients with MDR TB. Seventy-four (40%) of these 184 patients received treatment with linezolid. Treatment success rates ranged from 59% for the entire cohort (59% for MDR TB and 57% for XDR TB) to 87% for those with a definitive outcome (n = 125; 89% for MDR TB and 80% for XDR TB). Extensive drug susceptibility testing and availability of second- and third-line drugs under inpatient management conditions permit relatively high treatment success rates in MDR- and XDR TB.

Contrasting Effects of Ethylene Biosynthesis on Induced Plant Resistance against a Chewing and a Piercing-Sucking Herbivore in Rice

Ethylene is a stress hormone with contrasting effects on herbivore resistance. However, it remains unknown whether these differences are plant- or herbivore-specific. We cloned a rice 1-aminocyclopropane-1-carboxylic acid (ACC) synthase gene, OsACS2, whose transcripts were rapidly up-regulated in response to mechanical wounding and infestation by two important pests: the striped stem borer (SSB) Chilo suppressalis and the brown planthopper (BPH) Nilaparvata lugens. Antisense expression of OsACS2 (as-acs) reduced elicited ethylene emission, SSB-elicited trypsin protease inhibitor (TrypPI) activity, SSB-induced volatile release, and SSB resistance. Exogenous application of ACC restored TrypPI activity and SSB resistance. In contrast to SSB, BPH infestation increased volatile emission in as-acs lines. Accordingly, BPH preferred to feed and oviposit on wild-type (WT) plants—an effect that could be attributed to two repellent volatiles, 2-heptanone and 2-heptanol, that were emitted in higher amounts by as-acs plants. BPH honeydew excretion was reduced and natural enemy attraction was enhanced in as-acs lines, resulting in higher overall resistance to BPH. These results demonstrate that ethylene signaling has contrasting, herbivore-specific effects on rice defense responses and resistance against a chewing and a piercing-sucking insect, and may mediate resistance trade-offs between herbivores of different feeding guilds in rice.

Natural Variation in Maize Aphid Resistance Is Associated with 2,4-Dihydroxy-7-Methoxy-1,4-Benzoxazin-3-One Glucoside Methyltransferase Activity

Plants differ greatly in their susceptibility to insect herbivory, suggesting both local adaptation and resistance tradeoffs. We used maize (Zea mays) recombinant inbred lines to map a quantitative trait locus (QTL) for the maize leaf aphid (Rhopalosiphum maidis) susceptibility to maize Chromosome 1. Phytochemical analysis revealed that the same locus was also associated with high levels of 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one glucoside (HDMBOA-Glc) and low levels of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside (DIMBOA-Glc). In vitro enzyme assays with candidate genes from the region of the QTL identified three O-methyltransferases (Bx10a-c) that convert DIMBOA-Glc to HDMBOA-Glc. Variation in HDMBOA-Glc production was attributed to a natural CACTA family transposon insertion that inactivates Bx10c in maize lines with low HDMBOA-Glc accumulation. When tested with a population of 26 diverse maize inbred lines, R. maidis produced more progeny on those with high HDMBOA-Glc and low DIMBOA-Glc. Although HDMBOA-Glc was more toxic to R. maidis than DIMBOA-Glc in vitro, BX10c activity and the resulting decline of DIMBOA-Glc upon methylation to HDMBOA-Glc were associated with reduced callose deposition as an aphid defense response in vivo. Thus, a natural transposon insertion appears to mediate an ecologically relevant trade-off between the direct toxicity and defense-inducing properties of maize benzoxazinoids.