Development of carboxymethyl cellulose-chitosan hybrid micro- and macroparticles for encapsulation of probiotic bacteria.

Novel carboxymethyl cellulose-chitosan (CMC-Cht) hybrid micro- and macroparticles were successfully prepared in aqueous media either by drop-wise addition or via nozzle-spray methods. The systems were either physically or chemically crosslinked using genipin as the reticulation agent. The macroparticles (ca. 2mm) formed are found to be essentially of the core-shell type, while the microparticles (ca. 5µm) are apparently homogeneous. The crosslinked particles are robust, thermally resistant and less sensitive to pH changes. On the other hand, the physical systems are pH sensitive presenting a remarkable swelling at pH 7.4, while little swelling is observed at pH 2.4. Furthermore, model probiotic bacteria (Lactobacillus rhamnosus GG) was for the first time successfully encapsulated in the CMC-Cht based particles with acceptable viability count. Overall, the systems developed are highly promising for probiotic encapsulation and potential delivery in the intestinal tract with the purpose of modulating gut microbiota and improving human health.

Outbreak of KPC-3-producing ST15 and ST348 Klebsiella pneumoniae in a Portuguese hospital.

To date, only a few sporadic cases of infections due to Klebsiella pneumoniae carbapenemase (KPC) producers have been reported in Portugal. Here, we report for the first time an outbreak of K. pneumoniae KPC-3 producers in a tertiary-care hospital during 2013. Twenty-seven ertapenem-resistant K. pneumoniae were identified in patients at a tertiary-care hospital during 2013 isolated predominantly from urine (48·1%) and blood (25·9%) cultures. All isolates were highly resistant to ß-lactam antibiotics and most showed intermediate resistance to imipenem. The more frequent ß-lactamases were TEM- (77·7%), CTX-M- (70·3%) and KPC-type (66·6%). KPC-3 was identified by sequencing. The bla KPC-3 gene was associated with an IncF plasmid, and efficiently transferred to E. coli J53. Pulsed-field gel electrophoresis typing revealed three clusters of isolates which were further characterized by multi-locus sequence typing as ST11, ST15 and ST348. Ertapenem-resistant ST15 was already in circulation in the hospital, related to expression of OmpK36 modified porin, but the other two sequence types had not been previously found in the hospital. We conclude that the IncF plasmid mediated transfer of KPC-3 in the outbreak and that implementation of carbapenemase gene screening in isolates from patients on admission to hospital is advisable in order to control dissemination of these antimicrobial resistance elements.

HydroCalc Proteome: a tool to identify distinct characteristics of effector proteins.

Bacterial pathogenicity is associated with secretion of effector proteins into intra- and extracellular spaces. These proteins interfere with cellular processes such as inhibition of phagosome-lysosome fusion, induction of apoptosis and autophagy, activation and suppression of kinases, regulation of receptor activity, and modulation of transcription factors. Knowledge regarding the characteristics of these proteins would assist in pathogenicity studies, and help to identify possible and novel targets for antibacterial drugs. Amino acid hydropathy is a property that can affect behavior patterns in effector proteins. The HydroCalc Proteome tool analyzes total hydropathy, average hydropathy, C-terminal hydropathy, C-terminal load, and basic polar amino acids at the C-terminus. These five properties could contribute to the identification of proteins with an effector potential. HydroCalc Proteome is a web tool that provides a simple interface for the analysis of hydropathy properties in proteins. This tool permits the analysis of a single protein or even the complete proteome, which cannot be achieved by using other hydropathy tools. The tool displays the result of five properties related to effector proteins in a single table. The HydroCalc Proteome (www.gmb.bio.br/hydrocalc) is a powerful tool for protein analysis, and can contribute to the study of effector proteins.

Q Fever Dairy Herd Status Determination Based on Serological and Molecular Analysis of Bulk Tank Milk.

Ruminants are recognized as the main reservoirs of Coxiella burnetii. EFSA highlighted the lack of knowledge about Q fever prevalence in many European countries. A cross-sectional study was carried out in randomly selected dairy herds (n = 109) from central Portugal to screen for C. burnetii infection and to correlate it with herd factors. Bulk tank milk (BTM) samples from cattle (n = 45) and small ruminant (n = 64) herds were tested by ELISA and PCR. The apparent seroprevalence of Q fever was estimated in 45.9% (95% CI: 36.3-55.7) being higher in small ruminants (51.6; 95% CI: 39.6-63.4) than in cattle (37.8; 95% CI: 25.1-52.4). The shedding of C. burnetii in BTM was detected in 11.9% (95% CI: 7.1-19.4) of BTM, and it was higher in cattle (20%; 95% CI: 10.9-33.8) than in sheep and mixed herds (6.3%; 95% CI: 2.5-15). A high bacterial load (≥ 3 × 10(3) bacteria/ml) was observed in 85% of PCR-positive BTM. A significant correlation was found between the bacterial load and positive samples on ELISA (P < 0.001). Antibody positivity was significantly associated with the increased herd size (P < 0.01) and the occurrence of abortion (P < 0.05), whereas the shedding of C. burnetii was significantly associated with the report of infertility (P < 0.05). The results highlight that serological and molecular methods in combination are a useful tool to screen for Q fever and to clarify the herd infection status. The shedding of C. burnetii through milk is important, especially in dairy cattle, and thus, the role of milk as a potential source of infection among dairy workers should not be neglected. To our knowledge, this is the first study reporting C. burnetii infection in dairy livestock in Portugal showing that Q fever is significant in dairy herds, leading to economic losses and being a risk for public health, which highlights the need of implementation of control measures.

Intercalation and retention of carbon dioxide in a smectite clay promoted by interlayer cations.

A good material for CO2 capture should possess some specific properties: (i) a large effective surface area with good adsorption capacity, (ii) selectivity for CO2, (iii) regeneration capacity with minimum energy input, allowing reutilization of the material for CO2 adsorption, and (iv) low cost and high environmental friendliness. Smectite clays are layered nanoporous materials that may be good candidates in this context. Here we report experiments which show that gaseous CO2 intercalates into the interlayer nano-space of smectite clay (synthetic fluorohectorite) at conditions close to ambient. The rate of intercalation, as well as the retention ability of CO2 was found to be strongly dependent on the type of the interlayer cation, which in the present case is Li(+), Na(+) or Ni(2+). Interestingly, we observe that the smectite Li-fluorohectorite is able to retain CO2 up to a temperature of 35°C at ambient pressure, and that the captured CO2 can be released by heating above this temperature. Our estimates indicate that smectite clays, even with the standard cations analyzed here, can capture an amount of CO2 comparable to other materials studied in this context.

Characterization of a novel international clonal complex (CC32) of Acinetobacter baumannii with epidemic potential.

Twelve non-replicate Acinetobacter baumannii isolates from five European hospitals, Kuwait, and the US military healthcare system collected between 1980 and 2005 revealed a new clone, CC32. These included representative isolates of outbreaks/cross-infections. Antimicrobial susceptibility and carbapenem-resistant genetic traits varied. The widespread occurrence, the association with an outbreak and the carbapenem resistance indicate that CC32 has epidemic potential.

Serological evidence of exposure to Coxiella burnetii in sheep and goats in central Portugal.

The recent outbreak of Q fever in The Netherlands warned European health authorities of the need of studying Coxiella burnetii. In Portugal, little is known about C. burnetii infection in animals. A cross-sectional study was designed to investigate the exposure to C. burnetii in sheep and goats in the Central region of Portugal, estimating the herd and individual prevalence. A serosurvey was conducted in a two levels random sampling of 89 herds and 460 animals. Individual blood samples were collected from animals older than 6 months, and specific antibodies anti-C. burnetii were detected by ELISA testing. Results showed a global herd prevalence of 32.6% (95% CI: 23.1-42.1%). Herd prevalence was higher in mixed herds (38.5%; 95% CI: 12-65%) and in sheep herds (37.5%; 95% CI: 21-54%) than in goat herds (28.8%; 95% CI: 17-41%). Global individual prevalence was estimated at 9.6% (95% CI: 6.9-12.2%), and it was higher in goats (10.4%; 95% CI: 7.8-13%) than in sheep (8.6%; 95% CI: 5.8-11.4%). Sample positive percentages (S/P) ranged from 41.5% to 185.9%. S/P percent higher than 100 was found in 18.2% (8/44) of sera from distinct herds. Positive results were significantly associated with goats, older animals and larger herds. These results revealed the presence of C. burnetii in small ruminants evidencing their potential role in the infection cycle.

Long-term dissemination of an OXA-40 carbapenemase-producing Acinetobacter baumannii clone in the Iberian Peninsula.

OBJECTIVE: The main objectives of this study were to assess the clonal relatedness of Acinetobacter baumannii carbapenem-resistant isolates recovered from the Iberian Peninsula and to investigate the production of carbapenemases. METHODS: One hundred and sixty-two imipenem-resistant A. baumannii isolates were collected from 1998 to 2003 in three Portuguese university hospitals. An imipenem-resistant isolate (988FFP strain) recovered in 1995 from a smaller hospital unit, was also included, as well as an OXA-40-producing A. baumannii Spanish strain (SM28). Susceptibility tests were carried out by disc diffusion and Etest methods. DNA fingerprints were obtained by PFGE of ApaI-digested chromosomal DNA. Carbapenemase activity was determined by a bioassay and spectrophotometry. The detection of the blaOXA-40 gene was conducted through PCR analysis, cloning and nucleotide sequencing. RESULTS: All the isolates presented a similar multi-resistance pattern, including imipenem (MIC >32 mg/L). The Iberian isolates showed an identical PFGE pattern with minor band variations, including isolate 988FFP collected in 1995. PCR results revealed a blaOXA-type gene in 65 isolates and nucleotide sequence analysis revealed the presence of the blaOXA-40 gene in seven representative Portuguese isolates from the various geographically dispersed hospitals. CONCLUSIONS: Our results indicate that a multi-resistant epidemic clone of A. baumannii, carrying blaOXA-40, is disseminated in the Iberian Peninsula, persisting in Portugal since 1995.

Synchrotron x-ray scattering studies of water intercalation in a layered synthetic silicate.

Synchrotron x-ray diffraction studies were performed on a synthetic layered silicate Fluorohectorite clay. Diffraction patterns along the stacking direction were obtained in surface reflection and bulk transmission geometries on bulk pressed samples under controlled temperature and relative humidity. One-dimensional structure factors modeling the positions of the intercalant atoms have been obtained for three stable hydration states. From the narrow (00l) peak widths we conclude that well-crystallized domains consist of stacks of about 100 platelets, forming crystallites of the order of 0.1 microm thick. These crystallites have an orientational angular distribution of about 24 degrees around the stacking direction and represent the solid framework for microporosity in these samples.

Effects of exercise training on baroreflex control of the cardiovascular system.

Dynamic exercise training has been recommended as an antihypertensive therapy and as a way to modify the effects of many cardiovascular risk factors (Arakawa, 1993; Arroll and Beaglehole, 1992; Kelly and McClellan, 1994: see references 1-3 in the paper). However, the mechanisms underlying the blood-pressure lowering effect of chronic exercise are still poorly understood. It has been suggested that a decrease in sympathetic tone is one of the major effects elicited by chronic exercise on the cardiovascular system. The importance of the sympathetic component is confirmed in this review, since it was found that in spontaneously hypertensive rats (SHR) a marked decrease in sympathetic activity occurred after exercise training. Moreover, our findings suggest that this effect is mediated by improving the depressed baroreceptor function, which is, in part, responsible for the attenuation of the baroreflex sensitivity observed in the sedentary SHR (Krieger et al., 1998, 1999; see references 4 and 5 in the paper).

Exercise training increases baroreceptor gain sensitivity in normal and hypertensive rats.

Exercise training attenuates arterial hypertension and increases baroreflex sensitivity in spontaneous hypertension. However, no information exists regarding the portion of the baroreflex arch in which this attenuation takes place. We tested the hypothesis that exercise training increases the afferent pathway sensitivity of baroreflex control in both normotensive and spontaneously hypertensive rats (SHR). Arterial pressure and whole-nerve activity of the aortic baroreceptor (multifiber preparation) were evaluated in 30 male rats assigned to 4 groups: sedentary and exercise-trained normotensive rats and sedentary and exercise-trained SHR. Exercise training was performed on a motor treadmill, 5 d/wk for 60 minutes, gradually progressing toward a speed of 26.8 m/min. Exercise training reduced mean arterial pressure in conscious exercise-trained SHR (183+/-4 versus 165+/-7 mm Hg). The relation between changes in aortic baroreceptor discharge and changes in systolic arterial pressure increased significantly in exercise-trained normotensive rats (2.09+/-0.1 versus 1.44+/-0.1%/mm Hg) and exercise-trained SHR (0.92+/-0.1 versus 0.71+/-0.1%/mm Hg) compared with their respective sedentary rats. Likewise, the average aortic baroreceptor gain sensitivity (calculated by logistic equation) was significantly higher in exercise-trained normotensive rats (2.25+/-0.19 versus 1.77+/-0.03%/mm Hg) and exercise-trained SHR (1.07+/-0.04 versus 0.82+/-0.05%/mm Hg) compared with their respective sedentary control rats. In conclusion, exercise training increases aortic baroreceptor gain sensitivity in normotensive and SHR, thus improving baroreceptor sensitivity, which may result in a more efficient arterial pressure regulation by the baroreflexes.