A clindamycin acetylated derivative with reduced antibacterial activity inhibits articular hyperalgesia and edema by attenuating neutrophil recruitment, NF-κB activation and tumor necrosis factor-α production.

We recently demonstrated that clindamycin exhibits activities in acute and chronic models of pain and inflammation. In the present study, we investigated the effects of clindamycin and a clindamycin acetylated derivative (CAD) in models of acute joint inflammation and in a microbiological assay. Joint inflammation was induced in mice by intraarticular (i.a.) injection of zymosan or lipopolysaccharide (LPS). Clindamycin or CAD were administered via the intraperitoneal route 1 h before zymosan or LPS. Paw withdrawal threshold, joint diameter, histological changes, neutrophil recruitment, tumor necrosis factor-α (TNF-α) production and phosphorylation of the IκBα and NF-κB/p65 were evaluated. In vitro assays were used to measure the antibacterial activity of clindamycin and CAD and also their effects on zymosan-induced TNF-α production by RAW264.7 macrophages. Clindamycin exhibited activity against Staphylococcus aureus and Salmonella Typhimurium ATCC® strains at much lower concentrations than CAD. Intraarticular injection of zymosan or LPS induced articular hyperalgesia, edema and neutrophil infiltration in the joints. Zymosan also induced histological changes, NF-κB activation and TNF-α production. Responses induced by zymosan and LPS were inhibited by clindamycin (200 and 400 mg/kg) or CAD (436 mg/kg). Both clindamycin and CAD inhibited in vitro TNF-α production by macrophages. In summary, we provided additional insights of the clindamycin immunomodulatory effects, whose mechanism was associated with NF-κB inhibition and reduced TNF-α production. Such effects were extended to a clindamycin derivative with reduced antibacterial activity, indicating that clindamycin derivatives should be investigated as candidates to drugs that could be useful in the management of inflammatory and painful conditions.

Designing an optimized landscape restoration with spatially interdependent non-linear models.

Brazilian Atlantic Forest is a biodiversity hotspot drastically fragmented due to different land use practices. Our understanding on the impacts of fragmentation and restoration practices on ecosystem functionality significantly increased during the last decades. However, it is unknown to our knowledge how a precision restoration approach, integrated with landscape metrics, will affect the decision-making process of forest restoration. Here, we applied Landscape Shape Index and Contagion metrics in a genetic algorithm for planning forest restoration in watersheds at the pixel level. We evaluated how such integration may configure the precision of restoration with scenarios related to landscape ecology metrics. The genetic algorithm worked toward optimizing the site, shape, and size of forest patches across the landscape according to the results obtained in applying the metrics. Our results, obtained by simulations of scenarios, support aggregation of forest restoration zones as expected, with priority restoration areas indicated where most of the aggregation of forest patches occurs. Our optimized solutions for the study area (Santa Maria do Rio Doce Watershed) predicted an important improvement of landscape metrics (LSI = 44 %; Contagion/LSI = 73 %). Largest shifts are suggested based on LSI (i.e., three larger fragments) and Contagion/LSI (i.e., only one well-connected fragment) optimizations. Our findings indicate that restoration in an extremely fragmented landscape will promote a shift toward more connected patches and with reduction of the surface:volume ratio. Our work explores the use of genetic algorithms to propose forest restoration based on landscape ecology metrics in a spatially explicit innovative approach. Our results indicate that LSI and Contagion:LSI ratio may affect the choice concerning precise location of restoration sites based on forest fragments scattered in the landscape and reinforce the usefulness of genetic algorithms to yield an optimized-driven solution for restoration initiatives.

Influence of sucrose reduction on fouling during the production of dulce de leche.

In this Research Communication we focus the food industry´s broad tendency to decrease sugar content in food products onto dulce de leche (DL) and examine the influence of sucrose reduction on the detrimental deposits formed during the production process. The method used to identify the impact produced directly on the heat exchanger during the production of this product with low sucrose content required varying the quantity of sucrose in the milk. Different percentages of sucrose (20, 15, 10, 5 and 0% w/w) were submitted to the DL concentration process in a process simulator. After concentration, the quantification of the deposits formed in each was carried out and these deposits were characterized according to their composition. Methods such as Kjeldahl, Pregl-Dumas and sem-EDS were used. Thus, the work highlights the need to change the product manufacturing process due to changes in the formulation that directly impact the formation of deposits in the equipment used (fouling). This deposit changes significantly in relation to its quantity as well as in relation to the composition and chemical characteristics as the gradual reduction of the sucrose content in the production takes place. Therefore, these impacts must be considered in order to maintain better manufacturing and ensure efficient cleaning of equipment.

Presence and growth prediction of Staphylococcus spp. and Staphylococcus aureus in Minas Frescal cheese, a soft fresh cheese produced in Brazil.

Physical-chemical characteristics of Minas Frescal cheese (MFC) favor the growth of Staphylococcus spp. and allow the production of enterotoxins by specific strains. Here, we aimed to characterize the physical-chemical aspects (pH, storage temperature, and salt content) and the presence of Staphylococcus spp. in MFC samples (n = 50) to support a modeling study for the growth by this microorganism. Coagulase-positive staphylococci isolates were obtained and subjected to PCR assays to identify them as Staphylococcus aureus (nuc) and to detect staphylococcal enterotoxin-related genes (sea, seb, sec, sed, see). Staphylococcus aureus growth kinetics (maximum growth rate, Grmax, and lag time) were predicted based on ComBase model and MFC physical-chemical aspects. Mean counts of Staphylococcus spp. ranged from 3.3 to 6.7 log cfu/g, indicating poor hygiene practices during production. Selected isolates (n = 10) were identified as S. aureus, but none presented classical enterotoxin-related genes. pH, temperature, and salt content ranged from 5.80 to 6.62, 5°C to 12°C, and 0.85% to 1.70%, respectively. The Grmax values ranged from 0.012 to 0.419 log cfu/g per h. Independent of the storage temperature, the lowest Grmax values (0.012 to 0.372 log cfu/h) were obtained at pH 5.80 associated with salt content of 1.7%; independent of the pH and salt content, the best temperature to avoid staphylococcal growth was 7.5°C. Hygienic conditions during MFC production must be adopted to avoid staphylococcal contamination, and storage at temperatures lower than 7.5°C can prevent staphylococcal growth and the potential production of enterotoxins.

Interactions between caseins and food-derived bioactive molecules: A review.

Caseins are recognized as safe for consumption, abundant, renewable and have high nutritional value. Casein molecules are found in different aggregation states and their multiple binding sites offer the potential for delivering biomolecules with nutritional and/or health benefits, such as vitamins, phytochemicals, fibers, lipids, minerals, proteins, peptides, and pharmaceutical compounds. In the present review, we highlight the interactions between caseins and food-derived bioactive molecules, with a special focus on the aggregation states of caseins and the techniques used to produce and study the particles used for delivering. Research on interactions between caseins-minerals and casein-pharmaceutical molecules are not included here. This review aims to support the development of new and innovative functional foods in which caseins can be used as designed delivery systems.

Technological properties of beneficial bacteria from the dairy environment and development of a fermented milk with the beneficial strain Lactobacillus casei MRUV6.

In this research paper we describe the technological properties of beneficial lactic acid bacteria (LAB) obtained from a dairy production chain and the development of a fermented milk produced with Lactobacillus casei MRUV6. Fifteen LAB isolates (Lactobacillus sp., Pediococcus sp. and Weissela sp.) presented acidifying abilities (pH ranges from 0.73 to 2.11), were able to produce diacetyl (except by 5 isolates) and exopolysaccharides, and two were proteolytic. L. casei MRUV6 was selected for producing a fermented milk, stored up to 35 d at 4 and 10°C. Counts on MRS agar with added vancomycin (10 mg/l) and MRS agar with added bile salts (1.5% w/v) ranged from 9.7 to 9.9 log CFU/g, independently of the tested conditions, indicating stability and intestinal resistance of L. casei MRUV6, despite some significant differences (P < 0.05). The study demonstrated the technological potential of a potential probiotic candidate strain, L. casei MRUV6, to be used as a starter culture in the dairy industry.

Short communication: Potential use of passion fruit (Passiflora cincinnata) as a biopreservative in the production of coalho cheese, a traditional Brazilian cheese.

Passion fruit (Passiflora cincinnata Mast.) is a native fruit from the Caatinga, a typical ecoregion in northeastern Brazil, and it has potential for use by the food and pharmaceutical industries. In this study, we characterized the antimicrobial activity of P. cincinnata and its application in the production of coalho cheese, a traditional Brazilian product. Aqueous extract of P. cincinnata exhibited high inhibitory activity against Listeria spp. (n = 4, reference strains), Staphylococcus aureus (n = 3, reference strains), and multidrug-resistant Staph. aureus (n = 8), and low inhibitory activity against lactic acid bacteria (LAB, n = 3, reference strains). Based on these results, we produced coalho cheese using goat milk with and without (control) passion fruit. Cheeses were stored at 10°C for 14 d and populations of mesophilic aerobes, Staph. aureus, and presumptive LAB were monitored at d 1, 7 and 14. The passion fruit cheese had lower counts of mesophilic aerobes, Staph. aureus (after 7 and 14 d), and presumptive LAB (after 14 d) than the control cheese. Adding ground passion fruit contributed to a reduction of Staph. aureus counts in goat cheese, although these differences were not significant. These results indicated the inhibitory potential of passion fruit and its potential use for controlling microbial populations in a cheese model; further studies are needed to characterize the active molecules that are responsible for such activity.

Genetic Characterization of Some Neoponera (Hymenoptera: Formicidae) Populations Within the foetida Species Complex.

The foetida species complex comprises 13 Neotropical species in the ant genus Neoponera. Neoponera villosa Fabricius (1804) , Neoponera inversa Smith (1858), Neoponera bactronica Fernandes, Oliveira & Delabie (2013), and Neoponera curvinodis (Forel, 1899) have had an ambiguous taxonomic status for more than two decades. In southern Bahia, Brazil, these four species are frequently found in sympatry. Here we used Bayesian Inference and maximum likelihood analyses of COI and 16S mtDNA sequence data and conventional cytogenetic data together with observations on morphology to characterize sympatric populations of N. villosa, N. inversa, N. bactronica, and N. curvinodis. Our results showed marked differences in the karyotype of these ants. Both N. curvinodis and N. inversa have chromosome number of 2n = 30. Their chromosome composition, however, is distinct, which indicates that N. curvinodis is more closely related to N. bactronica. These four species clustered into three distinct groups. The close relationship between N. bactronica and N. curvinodis deserves further investigation since it has not been fully resolved here. Our results confirm that N. inversa, N. villosa, N. bactronica + N. curvinodis indeed represent four distinct taxa within the foetida species complex.

Physico-chemical and compositional analyses and 5-hydroxymethylfurfural concentration as indicators of thermal treatment intensity in experimental dulce de leche.

This Research Paper addresses the hypothesis that the type of dulce de leche formula affects formation of 5-hydroxymethylfurfural (HMF) during production and storage. Milk and sweetened condensed milk are products in which the Maillard reaction (MR) defines important characteristics such as colour and taste. There are few studies on the effects of pH, time, concentration, and sugar type on the formation of HMF or other MR markers in DL. Four formulas (varying in the addition of sodium bicarbonate and the type of sugar) were analysed for moisture, lipid, protein, ash, carbohydrate, water activity (Aw), and soluble solids. We found low variability in physicochemical and compositional attributes, but an elevation of HMF indices throughout the manufacturing. We determined that the addition of glucose and the use of relatively high concentrations of sodium bicarbonate caused these HMF indicator increases. These results inform DL research and production by the dairy industries and the scientific community, and highlight the importance of control in manufacturing.

pH- and ionic strength-dependent interaction between cyanidin-3-O-glucoside and sodium caseinate.

Understanding the mechanism of interaction between food proteins and bioactives constitutes the preliminary step to design food grade nanocarriers. We investigated the interaction between cyanidin-3-O-glucoside (C3G), and 20nm-sized sodium caseinate nanoparticles (NaCas) at pH 7 and pH 2 by fluorescence spectroscopy and dynamic light scattering. The characterization of the C3G-NaCas interaction indicated that the fluorescence quenching mechanism was predominantly static. C3G interacted with two sets of binding sites with association constants Ka of 106 and 105M-1. Electrostatic interactions dominated at pH 7, while hydrophobic effects were the main force at pH 2. Interestingly, the two sets of binding sites were discriminated by ionic strength at pH 7. The binding of C3G slightly modified the average diameter of NaCas nanoparticles without alteration of its surface charge suggesting a complexation of C3G molecules in the internal casein structure. Thus, NaCas constitutes a putative nanocarrier for anthocyanins in new functional foods.

The roles of barriers, refugia, and chromosomal clines underlying diversification in Atlantic Forest social wasps.

Phylogeographic studies have sought to explain the genetic imprints of historical climatic changes and geographic barriers within the Brazilian Atlantic Forest (AF) biota, and consequently two processes of diversification (refugia and barriers) have been proposed. Additionally, there is evidence that eustatic changes influenced the biogeographic history of the AF. Here we evaluate these contrasting diversification processes using two AF social wasp species - the mid-montane Synoeca cyanea and the lowland Synoeca aff. septentrionalis. We analyzed several sources of data including multilocus DNA sequence, climatic niche models and chromosomal features. We find support for idiosyncratic phylogeographic patterns between these wasps, involving different levels of population structure and genetic diversity, contrary suitable climatic conditions during the last glaciation, and contrasting historical movements along the AF. Our data indicate that neotectonics and refugia played distinct roles in shaping the genetic structure of these wasps. However, we argue that eustatic changes influenced the demographic expansion but not population structure in AF biota. Notably, these wasps exhibited chromosomal clines, involving chromosome number and decreasing of GC content, latitudinally oriented along the AF. Together, these results reinforce the need to consider individual organismal histories and indicate that barriers and refugia are significant factors in understanding AF evolution.

How the presence of a small molecule affects the complex coacervation between lactoferrin and ß-lactoglobulin.

Heteroprotein complex coacervation corresponds to the formation of two liquid phases in equilibrium induced by the interaction of two oppositely charged proteins. The more concentrated phase known as coacervate phase, has attracted interest from several fields of science due to its potential applications for example for encapsulation and delivery of bioactives. Prior such application, it is necessary to understand how the presence of small ligands affects the complex coacervation. In this work, we report on the interaction of small ligand with individual proteins ß-lactoglobulin (ß-LG) and lactoferrin (LF) and consequences on their complex coacervation. ANS (8-Anilinonaphthalene-1-sulfonic acid), a fluorescent probe, was used as model ligand. While ANS did not interact with ß-LG, it presented two sets of binding sites with LF inducing its self-aggregation. Depending on its concentration, ANS modulated the shape of ß-LG-LF macromolecular assembly. Coacervates were observed for ANS/LF molar ratio <25 against amorphous aggregates for higher ANS/LF molar ratios. A maximum loading capacity of around 40mg of ANS per gram of LF in the formed heteroprotein coacervates was reached.

Binding of Folic Acid Induces Specific Self-Aggregation of Lactoferrin: Thermodynamic Characterization.

In the study presented here, we investigated the interaction at pH 5.5 between folic acid (FA) and lactoferrin (LF), a positively charged protein. We found a binding constant Ka of 10(5) M(-1) and a high stoichiometry of 10 mol of FA/mol of LF. The size and charge of the complexes formed evolved during titration experiments. Increasing the ionic strength to 50 mM completely abolished the isothermal titration calorimetry (ITC) signal, suggesting the predominance of electrostatic interactions in the exothermic binding obtained. We developed a theoretical model that explains the complex triphasic ITC profile. Our results revealed a two-step mechanism: FA/LF interaction followed by self-association of the complexes thus formed. We suggest that 10 FA molecules bind to LF to form saturated reactive complexes (FA10/LF) that further self-associate into aggregates with a finite size of around 15 nm. There is thus a critical saturation degree of the protein, above which the self-association can take place. We present here the first results that provide comprehensive details of the thermodynamics of FA/LF complexation-association. Given the high stoichiometry, allowing a load of 55 mg of FA/g of LF, we suggest that FA/LF aggregates would be an effective vehicle for FA in fortified drinks.

Development of casein microgels from cross-linking of casein micelles by genipin.

Casein micelles are porous colloidal particles, constituted of casein molecules, water, and minerals. The vulnerability of the supramolecular structure of casein micelles face to changes in the environmental conditions restrains their applications in other domains besides food. Thus, redesigning casein micelles is a challenge to create new functionalities for these biosourced particles. The objective of this work was to create stable casein microgels from casein micelles using a natural cross-linker, named genipin. Suspensions of purified casein micelles (25 g L(-1)) were mixed with genipin solutions to have final concentrations of 5, 10, and 20 mM genipin. Covalently linked casein microgels were formed via cross-linking of lysyl and arginyl residues of casein molecules. The reacted products exhibited blue color. The cross-linking reaction induced gradual changes on the colloidal properties of the particles. The casein microgels were smaller and more negatively charged and presented smoother surfaces than casein micelles. These results were explained based on the cross-linking of free NH2 present in an external layer of κ-casein. Light scattering and rheological measurements showed that the reaction between genipin and casein molecules was intramicellar, as one single population of particles was observed and the values of viscosity (and, consequently, the volume fraction of the particles) were reduced. Contrary to the casein micelles, the casein microgels were resistant to the presence of dissociating agents, e.g., citrate (calcium chelating) and urea, but swelled as a consequence of internal electrostatic repulsion and the disruption of hydrophobic interactions between protein chains. The casein microgels did not dissociate at the air-solution interface and formed solid-like interfaces rather than a viscoelastic gel. The potential use of casein microgels as adaptable nanocarriers is proposed in the article.

UFV-P2 as a member of the Luz24likevirus genus: a new overview on comparative functional genome analyses of the LUZ24-like phages.

BACKGROUND: Phages infecting spoilage microorganisms have been considered as alternative biocontrol agents, and the study of their genomes is essential to their safe use in foods. UFV-P2 is a new Pseudomonas fluorescens-specific phage that has been tested for its ability to inhibit milk proteolysis. RESULTS: The genome of the phage UFV-P2 is composed of bidirectional modules and presented 75 functionally predict ORFs, forming clusters of early and late transcription. Further genomic comparisons of Pseudomonas-specific phages showed that these viruses could be classified according to conserved segments that appear be free from genome rearrangements, called locally collinear blocks (LCBs). In addition, the genome organization of the phage UFV-P2 was shown to be similar to that of phages PaP3 and LUZ24 which have recently been classified as a Luz24likevirus. CONCLUSIONS: We have presented the functional annotation of UFV-P2, a new Pseudomonas fluorescens phage. Based on structural genomic comparison and phylogenetic clustering, we suggest the classification of UFV-P2 in the Luz24likevirus genus, and present a set of shared locally collinear blocks as the genomic signature for this genus.

Complete Genome Sequence of the Pseudomonas fluorescens Bacteriophage UFV-P2.

Milk proteolysis caused by Pseudomonas fluorescens is a serious problem in the dairy industries as a result of its ability to grow under refrigeration. The use of phages to control contaminants in food has been considered an alternative to traditional methods; therefore, a thorough understanding of such organisms is vital for their use. In this study, we show the complete genome sequence and analysis of a P. fluorescens phage isolated from wastewater of a dairy industry in Brazil.

Cytogenetic characterization of two species of Frieseomelitta Ihering, 1912 (Hymenoptera, Apidae, Meliponini).

The cytogenetic analysis of Frieseomelitta dispar and F. francoi revealed the chromosome numbers 2n = 30 and n = 15 and a karyotypic formula 2K = 4M+2M(t)+4A+20A(M). The number of chromosomes observed was consistent with those reported for other Frieseomelitta species. The occurrence of the M(t) chromosome and other features of the karyotype formulae suggest a close relationship between F. dispar, F. francoi and F. varia. Nevertheless, it was possible to differentiate the karyotypes of the species by DAPI/CMA(3) staining, which revealed GC-rich regions on two chromosome pairs of F. dispar: one acrocentric and one pseudoacrocentric. In F. francoi, the same kinds of regions were observed on a pair of metacentrics and on a pair of acrocentrics. Our analysis also confirmed the chromosome number conservation in Frieseomelitta and suggests that infrequent pericentric inversion could constitute a synapomorphy for the group including F. dispar, F. francoi, and F. varia.

First records of Synoeca septentrionalis Richards, 1978 (Hymenoptera, Vespidae, Epiponini) in the Brazilian Atlantic Rain Forest.

Nests of Synoeca septentrionalis were collected in two Brazilian Atlantic Rain Forest localities (Itabuna and Santa Terezinha, in the state of Bahia and Alfredo Chaves in the state of Espírito Santo). Synoeca septentrionalis was previously recorded only from Central America and northwestern South America. This findingextends its geographical distribution to Northeast and Southeast regions of Brazil, and represents the first record for Synoeca septentrionalis in the Brazilian Atlantic Rain forest, raising to three the number of Synoeca species known from Bahia State.

Cytogenetic characterization of two species of Frieseomelitta Ihering, 1912 (Hymenoptera, Apidae, Meliponini)

The cytogenetic analysis of Frieseomelitta dispar and F. francoi revealed the chromosome numbers 2n = 30 and n = 15 and a karyotypic formula 2K = 4M+2Mt+4A+20A M. The number of chromosomes observed was consistent with those reported for other Frieseomelitta species. The occurrence of the Mt chromosome and other features of the karyotype formulae suggest a close relationship between F. dispar, F. francoi and F. varia. Nevertheless, it was possible to differentiate the karyotypes of the species by DAPI/CMA3 staining, which revealed GC-rich regions on two chromosome pairs of F. dispar: one acrocentric and one pseudoacrocentric. In F. francoi, the same kinds of regions were observed on a pair of metacentrics and on a pair of acrocentrics. Our analysis also confirmed the chromosome number conservation in Frieseomelitta and suggests that infrequent pericentric inversion could constitute a synapomorphy for the group including F. dispar, F. francoi, and F. varia.