Role of the mitogen-activated protein kinase pathway in the differential response of bovine monocytes to Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium

We compared the kinetics of activation and antimicrobial activities of MAPK-p38 and MAPK-ERK in bovine monocytes infected with Mycobacterium avium subsp. paratuberculosis (MAP) and Mycobacterium avium subsp. avium (Maa). Monocytes were incubated with MAP or Maa organisms with or without a specific inhibitor of the MAPK-p38 pathway (SB203580), and MAPK phosphorylation and antimicrobial functions of monocytes were evaluated. At early time points MAPK-p38 phosphorylation was greater in MAP-infected bovine monocytes than in Maa-infected monocytes. At later time points MAPK-p38 phosphorylation by both organisms was similar. MAPKp38 phosphorylation in MAP-infected monocytes was similar to negative control cells, whereas in Maa-infected this activation remained greater than negative control cells. Increase phosphorylation MAPK-ERK was similar at all time points for both organisms. Bovine monocytes had minimal capacity to kill MAP organisms, to acidify MAP-containing phagosomes, or to form phagolysosome. Alternatively, bovine monocytes were able to kill Maa organisms. Addition of SB203580 to monocyte cultures increased phagosome acidification, phagolysosome formation, and killing of MAP and Maa organisms. Taken together these data indicate that early transient activation of MAPK-p38 in bovine mononuclear phagocytes by MAP organisms may be a key mechanism involved in the capacity of MAP to survive in bovine monocytes.     

Harmonia axyridis: What will stop the invader?

In recent years Harmonia axyridis (Pallas, 1773) (Coleoptera: Coccinellidae) has become a very popular insect among biological control practitioners and scientists, not only for its potential to be an efficient biological control agent but also because it is considered invasive. Individuals of this species were deliberately introduced into several countries for biological control of different arthropods pests. However the predator itself became an invasive species, affecting the dynamics and composition of several guilds through direct or indirect interactions with established species, including intraguild predation. In this paper we discuss the reasons why the species has a high invasiveness and what are the limits to invasion by this species. It is not clear if the invasiveness of the beetle is linked to its biological, ecological and behavioural abilities, or to other factors such as invasibility and interactions between the invaders, the noninvaders, and the habitat, which may in part explain the reasons of its success and help us to answer the question “what will stop the invader?” We also discuss the reason for the absence of the predator in the Azores islands. Despite the intentional introduction of H. axyridis in the Azores and the high number of individuals released, there are no records of this species in the wild, despite recent extensive sampling effort. In this paper we discuss the reasons for the apparent failure or the delay in establishment of the predator. One factor which may hamper the establishment of H. axyridis in some of the Azores islands is the absence of winter environmental conditions, mainly the temperature which is seldom lower than 12°C, essential for the induction of diapause. The lack of success in the establishment could be also related to functional diversity saturation, that is species saturation and competitive exclusion of H. axyridis by other previously established species may be operating.     

Retail Survey of Brazilian Milk and Minas Frescal Cheese and a Contaminated Dairy Plant To Establish Prevalence, Relatedness, and Sources of Listeria monocytogenes Isolates

A study was designed to recover Listeria monocytogenes from pasteurized milk and Minas frescal cheese (MFC) sampled at retail establishments (REs) and to identify the contamination source(s) of these products in the corresponding dairy processing plant. Fifty milk samples (9 brands) and 55 MFC samples (10 brands) were tested from REs located in Juiz de Fora, Minas Gerais, Brazil. All milk samples and 45 samples from 9 of 10 MFC brands tested negative for L. monocytogenes; however, “brand F” of MFC obtained from REs 119 and 159 tested positive. Thus, the farm/plant that produced brand F MFC was sampled; all samples from the milking parlor tested negative for L. monocytogenes, whereas several sites within the processing plant and the MFC samples tested positive. All 344 isolates recovered from retail MFC, plant F MFC, and plant F environmental samples were serotype 1/2a and displayed the same AscI or ApaI fingerprints. Since these results established that the storage coolers served as the contamination source of the MFC, plant F was closed so that corrective renovations could be made. Following renovation, samples from sites that previously tested positive for the pathogen were collected from the processing environment and from MFC on multiple visits; all tested negative for L. monocytogenes. In addition, on subsequent visits to REs 159 and 119, all MFC samples tested negative for the pathogen. Studies are ongoing to quantify the prevalence, levels, and types of L. monocytogenes in MFC and associated processing plants to lessen the likelihood of listeriosis in Brazil.     

Efficacy of curcumin in the treatment of experimental vulvovaginal candidiasis

BackgroundCandida albicans is the main agent that causes vulvovaginal candidiasis. Resistance among isolates to azole antifungal agents has been reported.AimsDue to the well-known antifungal potential of curcumin, the purpose of this work was to evaluate the in vitro anticandidal activity of curcumin and its effect in the treatment of experimental vulvovaginal candidiasis.MethodsThe anticandidal activity of curcumin was investigated against eight Candida strains by the broth microdilution assay, and its mechanism of action was evaluated by testing the binding to ergosterol. Then, the effect of curcumin in the treatment of vulvovaginal candidiasis was evaluated in an immunosuppressed, estrogen treated rat model.ResultsCurcumin showed minimum inhibitory concentration values of 125–1000 μg/ml, and the best result was observed against Candida glabrata. The compound was shown to be able to bind to the ergosterol present in the membrane, event that may be the mechanism of action. In addition, in the in vivo model of vulvovaginal candidiasis with C. albicans, treatments reduced the vaginal fungal burden in infected rats after seven days of treatment with different doses.ConclusionsCurcumin could be considered a promising effective antifungal agent in the treatment of vulvovaginal candidiasis.     

Antinoceptive and Anti-inflammatory Activities of the Ethanolic Extract,Fractions and Flavones Isolated from Mimosa tenuiflora (Willd.) Poir (Leguminosae)

The bark of Mimosa tenuiflora (Willd.) Poiret (Leguminosae family), popularly known as “jurema preta” in Brazil, is used by the population of Contendas of Sincorá (Bahia State, Brazil) for the treatment of coughs and wound healing. Thus, the aim of this study was to evaluate the antinociceptive and anti-inflammatory activities of the bark ethanol extract (EEMT) and solvent soluble fractions (hexane—H, DCM—D, EtOAc—E and BuOH—B) of the extract in vivo. Additionally, we synthesized 5,7-dihidroxy-4’-methoxyflavanone (isosakuranetin) and isolated the compound sakuranetin, and both compounds were also tested. The anti-inflammatory and antinociceptive assays performed were: writhing test; nociception induced by intraplantar formalin injection; leukocyte recruitment to the peritoneal cavity; evaluation of vascular permeability (Evans blue test); and evaluation of mechanical hypernociception (von Frey test). Production of TNF-α, IL-10, myeloperoxidase and the expression of ICAM-1 were also evaluated. Statistical analysis was performed by one-way ANOVA followed by the Bonferroni post-test (n = 8), with P < 0.05. The EEMT showed antinociceptive activities in writhing test (100–200 mg/kg), in the second phase of the formalin test (50–200 mg/kg), and in mechanical hypernociception (100 mg/kg). EEMT showed an anti-inflammatory effect by reducing neutrophil migration to the peritoneal cavity and in the plantar tissue detected by the reduction of myeloperoxidase activity (100 mg/kg), reduction of IL-10 levels and expression of ICAM-1 in the peritoneal exudate and the mesentery (100 mg/kg), respectively. The four soluble EEMT fractions showed good results in tests for antinociceptive (H, D, E, B) and anti-inflammation (H, D, E). Only sakuranetin showed reduction of the writhing and neutrophil migration (200 mg/kg). Thus, the EEMT and soluble fractions of M. tenuiflora bark demonstrated great antinociceptive and anti-inflammatory activities, as also sakuranetin. More studies should be conducted to elucidate the mechanism of action of this compound. To the best of our knowledge, this is the first report on the antinociceptive activity of the M. tenuiflora fractions and the bioactive isolated compound sakuranetin in vivo.     

Focus on Metabolism: Changes in Whole-Plant Metabolism during the Grain-Filling Stage in Sorghum Grown under Elevated CO2 and Drought

Projections indicate an elevation of the atmospheric CO₂ concentration ([CO₂]) concomitant with an intensification of drought for this century, increasing the challenges to food security. On the one hand, drought is a main environmental factor responsible for decreasing crop productivity and grain quality, especially when occurring during the grain-filling stage. On the other hand, elevated [CO₂] is predicted to mitigate some of the negative effects of drought. Sorghum (Sorghum bicolor) is a C₄ grass that has important economical and nutritional values in many parts of the world. Although the impact of elevated [CO₂] and drought in photosynthesis and growth has been well documented for sorghum, the effects of the combination of these two environmental factors on plant metabolism have yet to be determined. To address this question, sorghum plants (cv BRS 330) were grown and monitored at ambient (400 µmol mol⁻¹) or elevated (800 µmol mol⁻¹) [CO₂] for 120 d and subjected to drought during the grain-filling stage. Leaf photosynthesis, respiration, and stomatal conductance were measured at 90 and 120 d after planting, and plant organs (leaves, culm, roots, prop roots, and grains) were harvested. Finally, biochemical composition and intracellular metabolites were assessed for each organ. As expected, elevated [CO₂] reduced the stomatal conductance, which preserved soil moisture and plant fitness under drought. Interestingly, the whole-plant metabolism was adjusted and protein content in grains was improved by 60% in sorghum grown under elevated [CO₂].Global food demand is projected to increase up to 110% by the middle of this century (Tilman et al., 2011; Alexandratos and Bruinsma, 2012), particularly due to a rise in world population that is likely to plateau at about 9 billion people (Godfray et al., 2010). Additionally, the average concentration of atmospheric CO₂ ([CO₂]) has increased 1.75 µmol mol⁻¹ per year between 1975 and today, reaching 400 µmol mol⁻¹ in April 2015 (NOAA, 2015). According to the A2 emission scenario from the U.S. Environmental Protection Agency, in the absence of explicit climate change policy, atmospheric CO₂ concentrations will reach 800 µmol mol⁻¹ by the end of this century. The increasing atmospheric [CO₂] is resulting in global climate changes, such as reduction in water availability and elevation in temperature. These factors are expected to heavily influence food production in the next years (Godfray and Garnett, 2014; Magrin et al., 2014).Sorghum (Sorghum bicolor) is a C₄ grass, considered a staple food grain for millions of the poorest and most food-insecure people in the semiarid tropics of Africa, Asia, and Central America, serving as an important source of energy, proteins, vitamins, and minerals (Taylor et al., 2006). Moreover, this crop is used for animal feed and as industrial raw material in developed countries such as the United States, which is the main world producer (FAO, 2015). With a fully sequenced genome (Paterson et al., 2009) and over 45,000 accessions representing a large geographic and genetic diversity, sorghum is a good model system in which to study the impact of global climate changes in C₄ grasses.The increase in [CO₂] in the atmosphere, which is the main driver of global climate changes (Meehl et al., 2007), is predicted to boost photosynthesis rates and productivity in a series of C₃ legumes and cereals, mainly due to a decrease in the photorespiration process (Grashoff et al., 1995; Long et al., 2006). On the contrary, due to their capacity to concentrate CO₂ in bundle sheath cells and reduce photorespiration to virtually zero, C₄ plants are unlikely to respond to the elevation of atmospheric [CO₂] (Leakey, 2009). However, even for C₄ plants, elevated [CO₂] can ameliorate the effects caused by drought, maintaining higher photosynthetic rates. This is due to an improvement in the efficiency of water use that is achieved by the reduction in stomatal conductance (Leakey et al., 2004; Markelz et al., 2011).The rate of photosynthesis as well as the redistribution of photoassimilates accumulated in different plant tissues during the day and/or during vegetative growth are crucial to grain development, and later, to its filling (Schnyder, 1993). Due to this relationship, any environmental stress such as drought occurring during the reproductive phase has the potential to result in poor grain filling and losses in yield (Blum et al., 1997). For instance, postanthesis drought can cause up to 30% decrease in yield (Borrell et al., 2000). It is also known that elevated [CO₂], drought, high temperature, and any combinations of these stresses can lead to significant changes in grain composition (Taub et al., 2008; Da Matta et al., 2010; Uprety et al., 2010; Madan et al., 2012), suggesting diverse metabolic alterations and/or adaptations that occur in the plant when it is cultivated in such conditions.Although the impacts of elevated [CO₂] and drought on photosynthesis and the growth of sorghum have been well documented (Conley et al., 2001; Ottman et al., 2001; Wall et al., 2001), no attention has been given to the impact of the combination of these two environmental changes on plant metabolism and composition. Regarding physiology, studies on the growth of sorghum under elevated [CO₂] and drought showed an increase of the net assimilation rate of 23% due to a decrease of 32% in stomatal conductance (Wall et al., 2001). This resulted in sorghum’s ability to use water 17% more efficiently (Conley et al., 2001). An improvement in the final overall biomass under elevated [CO₂] and drought has also been described (Ottman et al., 2001), but without a significant effect in grain yield (Wall et al., 2001).Few studies have been monitoring metabolic pathways in plants under elevated [CO₂] (Li et al., 2008; Aranjuelo et al., 2013) and drought (Silvente et al., 2012; Nam et al., 2015; Wenzel et al., 2015). Furthermore, to our knowledge, there are only two reports in which metabolite profiles or metabolic pathways were investigated under the combination of these two environmental conditions (Sicher and Barnaby, 2012; Zinta et al., 2014). Although it is widely accepted that whole-plant metabolism and composition can impact grain filling and yield, metabolic studies conducted so far have focused on a specific plant organ. For instance, Sicher and Barnaby (2012) analyzed the metabolite profile of leaves from maize (Zea mays) plants that were grown under elevated [CO₂] and drought, but they did not show how those environmental changes could have affected the metabolism of other tissues (e.g. culm and roots) or how they might have influenced the biomass or grain composition.In order to address how the combination of elevated [CO₂] and drought can modify whole-plant metabolism as well as biomass composition in sorghum, this study aimed to (1) evaluate photosynthesis, growth, and yield; (2) underline the differences in biomass composition and primary metabolite profiles among leaves, culm, roots, prop roots, and grains; and (3) determine the effect of elevated [CO₂] and drought on the primary metabolism of each organ.     

In vivo and in silico anti-inflammatory mechanism of action of the semisynthetic (−)-cubebin derivatives (−)-hinokinin and (−)-O-benzylcubebin

(?)-Cubebin (CUB), isolated from seeds of Piper cubeba, was used as starting material to obtain the derivatives (?)-hinokinin (HK) and (?)-O-benzyl cubebin (OBZ). Using paw edema as the experimental model and different chemical mediators (prostaglandin and dextran), it was observed that both derivatives were active in comparison with both negative (5% Tween® 80 in saline) and positive (indomethacin) controls. The highest reduction in the prostaglandin-induced edema was achieved by OBZ (66.0%), while HK caused a 59.2% reduction. Nonetheless, the dextran-induced paw edema was not significantly reduced by either of the derivatives (HK or OBZ), which inhibited edema formation by 18.3% and 3.5%, respectively, in contrast with the positive control, cyproheptadine, which reduced the edema by 56.0%. The docking analysis showed that OBZ presented the most stable ligand-receptor (COX-2 – cyclooxygenase-2) interaction in comparison with CUB and HK.     

The interaction of Trichomonas vaginalis and Tritrichomonas foetus with epithelial cells in vitro

The Madin-Darby canine kidney epithelial cell line (MDCK) was used as a model for trichomonad-host cell interaction. Two laboratory strains of the human parasite Trichomonas vaginalis and the cattle's parasite Tritrichomonas foetus or their supernatants from axenic cultures were allowed to interact with confluent epithelial cultures. The interaction process studied by transmission and scanning electron microscopy revealed that both parasites adhere to monolayers through flagella, cell body and particularly for T. foetus, through the posterior projection of the axostyle. A close contact region between the trichomonad's surface and MDCK cells was observed. A study of the involvement of trichomonad surface component in the interaction process indicated that cytochalasin B treated-parasites adhere much less to epithelial monolayers than untreated parasites. Colchicine treatment did not affect such adhesion. Treatment of the parasites with trypsin reduced the adhesion of trichomonads to monolayers but did not interfere with the cytopathic effect. In contrast, treatment of the parasites with neuraminidase did not interfere with their adhesion to epithelial cells and the monolayer destruction was further increased.     

Anti-Sporothrix Antibody Detection in Domestic Cats as an Indicator of a Possible New Occurrence Area for Sporotrichosis in North Brazil

Mycopathologia - Feline sporotrichosis has emerged as an important public health issue in some countries, especially Brazil. Currently, zoonotic transmission of Sporothrix brasiliensis by domestic...