Interactive impact of residual pyrethroid compounds used in the Chilean salmon farming industry and coastal acidification conditions on the feeding performance of farmed mussels in northern Patagonia.

The use of pyrethroids in aquaculture has been an important component of achieving a thriving salmon farming industry in Chile. While the residual presence of such substances is known to depend on environmental conditions, most ecotoxicological studies to date have not considered environmental context. Here, we conducted oceanographic monitoring combined with experiments aiming to estimate the effects of two pyrethroids on the feeding rates of larvae of farmed mussels, Mytilus chilensis. In additional experiments, mussel spats were exposed to both pyrethroids, but under contrasting temperature/pH so as to mimic winter and summer conditions. Experiments mimicking spring conditions revealed that both pyrethroid substances affected the feeding of mussel larvae as a function of concentration. Conversely, significant impact of pyrethroids on adults were not observed with regard to temperature and pH, but a significant impact of low temperature/low pH condition on ingestion rates was confirmed. Given the current status of increasing ocean acidification, the results of this study are expected to provide useful information with regard to achieving sustainable mussel aquaculture, especially considering both activities occur in similar geographic areas, and the expansion of salmon farming areas is ongoing in Chile.

Role of boron and its interaction with other elements in plants.

Boron (B) is an essential microelement for plants, and its deficiency can lead to impaired development and function. Around 50% of arable land in the world is acidic, and low pH in the soil solution decreases availability of several essential mineral elements, including B, magnesium (Mg), calcium (Ca), and potassium (K). Plants take up soil B in the form of boric acid (H3BO3) in acidic soil or tetrahydroxy borate [B(OH)4]- at neutral or alkaline pH. Boron can participate directly or indirectly in plant metabolism, including in the synthesis of the cell wall and plasma membrane, in carbohydrate and protein metabolism, and in the formation of ribonucleic acid (RNA). In addition, B interacts with other nutrients such as Ca, nitrogen (N), phosphorus (P), K, and zinc (Zn). In this review, we discuss the mechanisms of B uptake, absorption, and accumulation and its interactions with other elements, and how it contributes to the adaptation of plants to different environmental conditions. We also discuss potential B-mediated networks at the physiological and molecular levels involved in plant growth and development.

Improving Saccharomyces cerevisiae acid and oxidative stress resistance using a prokaryotic gene identified by functional metagenomics.

Innovations in obtaining products from lignocellulosic biomass have been largely based on the improvement of microorganisms and enzymes capable of degrading these materials. To complete the whole process, microorganisms must be able to ferment the resulting sugars and tolerate high concentrations of product, osmotic pressure, ion toxicity, temperature, toxic compounds from lignocellulose pretreatment, low pH, and oxidative stress. In this work, we engineered laboratory and industrial Saccharomyces cerevisiae strains by combining a gene (hu) recovered from a metagenomic approach with different native and synthetic promoters to obtain improved acid and oxidative stress resistance. Laboratorial strains harboring hu gene under the control of the synthetic stress responsive PCCW14v5 showed increased survival rates after 2 h exposure to pH 1.5. The hu gene was also able to significantly enhance the tolerance of the industrial strain to high concentrations of H2O2 when combined with PTEF1, PYGP1 or PYGP1v7 after 3 h exposure.

Extracellular acidosis stimulates breast cancer cell motility through aryl hydrocarbon receptor and c-src kinase activation.

A reduction in extracellular pH (pHe) is a characteristic of most malignant tumors. The aryl hydrocarbon receptor (AhR) is a transcription factor localized in a cytosolic complex with c-Src, which allows it to trigger nongenomic effects through c-Src. Considering that the slightly acidic tumor microenvironment promotes breast cancer progression in a similar way to the AhR/c-Src axis, our aim was to evaluate whether this pathway could be activated by low pHe. We examined the effect of pHe 6.5 on AhR/c-Src axis using two breast cancer cell lines (MDA-MB-231 and LM3) and mammary epithelial cells (NMuMG) and found that acidosis increased c-Src phosphorylation only in tumor cells. Moreover, the presence of AhR inhibitors prevented c-Src activation. Low pHe reduced intracellular pH (pHi), while amiloride treatment, which is known to reduce pHi, induced c-Src phosphorylation through AhR. Analyses were conducted on cell migration and metalloproteases (MMP)-2 and -9 activities, with results showing an acidosis-induced increase in MDA-MB-231 and LM3 cell migration and MMP-9 activity, but no changes in NMuMG cells. Moreover, all these effects were blocked by AhR and c-Src inhibitors. In conclusion, acidosis stimulates the AhR/c-Src axis only in breast cancer cells, increasing cell migration and MMP-9 activity. Although the AhR activation mechanism still remains elusive, a reduction in pHi may be thought to be involved. These findings suggest a critical role for the AhR/c-Src axis in breast tumor progression stimulated by an acidic microenvironment.

Effect of low pH on growth and shell mechanical properties of the Peruvian scallop Argopecten purpuratus (Lamarck, 1819).

Dissolution of anthropogenic CO2 modifies seawater pH, leading to ocean acidification, which might affect calcifying organisms such as bivalve mollusks. Along the Peruvian coast, however, natural conditions of low pH (7.6-8.0) are encountered in the habitat of the Peruvian scallop (Argopecten purpuratus), as a consequence of the nearby coastal upwelling influence. To understand the effects of low pH in a species adapted to these environmental conditions, an experiment was performed to test its consequences on growth, calcification, dissolution, and shell mechanical properties in juvenile Peruvian scallops. During 28 days, scallops (initial mean height = 14 mm) were exposed to two contrasted pH conditions: a control with unmanipulated seawater presenting pH conditions similar to those found in situ (pHT = 7.8) and a treatment, in which CO2 was injected to reduce pH to 7.4. At the end of the experiment, shell height and weight, and growth and calcification rates were reduced about 6%, 20%, 9%, and 10% respectively in the low pH treatment. Mechanical properties, such as microhardness were positively affected in the low pH condition and crushing force did not show differences between pH treatments. Final soft tissue weights were not significantly affected by low pH. This study provides evidence of low pH change shell properties increasing the shell microhardness in Peruvian scallops, which implies protective functions. However, the mechanisms behind this response need to be studied in a global change context.

Examination of ionoregulatory characteristics of South American cichlids.

We examined ionoregulatory traits of four cichlid species from South America, oscars (Astronotus ocellatus), Tapajos cichlids (Geophagus sp.), Macmaster's dwarf cichlids (Apistogramma macmasteri), and keyhole cichlids (Cleithracara maronii), all inhabitants of ion-poor waters. Km values for Na+ transport in fish held in 100 µmol L-1 Na+ water ranged from 49 to 143 µmol L-1, and Jmax values spanned 450 to 1205 nmol g-1 h-1. After one month in 1000 µmol L-1 Na+ water, kinetic parameters for Na+ uptake in three of the four species acclimated, but only oscars displayed the "typical" pattern of higher Km and lower Jmax values. Low pH water inhibited Na+ uptake (JinNa) in all, and stimulated Na+ efflux (JoutNa) 2.5 to 3.5-fold in three of the four species. Oscars alone had had a measurable JinNa at pH 3.5 and no stimulation of JoutNa. We measured JinNa in oscars and keyhole cichlids during exposure to 100 µmol L-1 Ethoxzolemide (EZ), an inhibitor of carbonic anhydrase, and 1 mmol L-1 NH4Cl (HEA). EZ inhibited JinNa by about 50% and HEA reduced JinNa by 80-90%. These results suggest that Na+ uptake involves H+ extrusion. Acute exposure to 1 µmol L-1 CuSO4 and 60 nmol L-1 AgNO3 inhibited JinNa in both species by 30-85%. Exposure of oscars to 5-fold higher concentrations of both metals did not further inhibit JinNa, but it did stimulate JoutNa 50-150%. The response to metals of both species are similar to other species that have been examined.

Extracellular Acidosis and mTOR Inhibition Drive the Differentiation of Human Monocyte-Derived Dendritic Cells.

During inflammation, recruited monocytes can differentiate either into macrophages or dendritic cells (DCs); however, little is known about the environmental factors that determine this cell fate decision. Low extracellular pH is a hallmark of a variety of inflammatory processes and solid tumors. Here, we report that low pH dramatically promotes the differentiation of monocytes into DCs (monocyte-derived DCs [mo-DCs]). This process is associated with a reduction in glucose consumption and lactate production, the upregulation of mitochondrial respiratory chain genes, and the inhibition of mTORC1 activity. Interestingly, we also find that both serum starvation and pharmacological inhibition of mTORC1 markedly promote the differentiation of mo-DCs. Our study contributes to better understanding the mechanisms that govern the differentiation of monocytes into DCs and reveals the role of both extracellular pH and mTORC1 as master regulators of monocyte cell fate.

Survival of microencapsulated Lactobacillus casei (prepared by vibration technology) in fruit juice during cold storage

Background: Foods including probiotics are considered "functional foods." As an alternative to dairy products, we investigated the behavior of Lactobacillus casei when exposed to low-pH fruit juice. Juices of fruits such as pineapple, raspberry, and orange were assessed. Free and microencapsulated forms of L. casei were compared, and the viability of the probiotic was evaluated under storage at 4°C for 28 d. Microbiological analyses were carried out to ensure a safe and healthy product for consumers who look for foods with probiotics from sources other than dairy. Results: Low pH affected L. casei survival during storage depending on the type of fruit juice. In the case of pineapple juice, some microcapsules were broken, but microcapsules recovered at the end of the storage period had 100% viability (2.3 × 107 CFU/g spheres). In the case of orange juice, more than 91% viability (5.5 × 106 CFU/g spheres) was found. In raspberry juice, viability decreased rapidly, disappearing at the end of the storage period, which was caused by the absorption of high concentrations of anthocyanin inside microcapsules more than low pH. Conclusion: Low pH affected the survival of L. casei under refrigeration; even when they were microencapsulated, acidic conditions impacted their viability. Although pH affects viability, its value is very sensitive and will depend on the type of fruit juice and its composition. Some fruit juices contain compounds used as substrates for Lactobacillus and other compounds with antimicrobial effects.

Effect of CO2 Concentration on Uptake and Assimilation of Inorganic Carbon in the Extreme Acidophile Acidithiobacillus ferrooxidans.

This study was motivated by surprising gaps in the current knowledge of microbial inorganic carbon (Ci) uptake and assimilation at acidic pH values (pH < 3). Particularly striking is the limited understanding of the differences between Ci uptake mechanisms in acidic versus circumneutral environments where the Ci predominantly occurs either as a dissolved gas (CO2) or as bicarbonate (HCO3 -), respectively. In order to gain initial traction on the problem, the relative abundance of transcripts encoding proteins involved in Ci uptake and assimilation was studied in the autotrophic, polyextreme acidophile Acidithiobacillus ferrooxidans whose optimum pH for growth is 2.5 using ferrous iron as an energy source, although they are able to grow at pH 5 when using sulfur as an energy source. The relative abundance of transcripts of five operons (cbb1-5) and one gene cluster (can-sulP) was monitored by RT-qPCR and, in selected cases, at the protein level by Western blotting, when cells were grown under different regimens of CO2 concentration in elemental sulfur. Of particular note was the absence of a classical bicarbonate uptake system in A. ferrooxidans. However, bioinformatic approaches predict that sulP, previously annotated as a sulfate transporter, is a novel type of bicarbonate transporter. A conceptual model of CO2 fixation was constructed from combined bioinformatic and experimental approaches that suggests strategies for providing ecological flexibility under changing concentrations of CO2 and provides a portal to elucidating Ci uptake and regulation in acidic conditions. The results could advance the understanding of industrial bioleaching processes to recover metals such as copper at acidic pH. In addition, they may also shed light on how chemolithoautotrophic acidophiles influence the nutrient and energy balance in naturally occurring low pH environments.

Room Temperature Growth of Salmonella enterica Serovar Saintpaul in Fresh Mexican Salsa.

Salsa-associated outbreaks, including the large multistate outbreak in the United States in 2008 caused by jalapeño and serrano peppers contaminated with Salmonella Saintpaul, have raised concerns about salsa as a potential vehicle for transmission. Despite these events, there has been relatively limited research on the potential growth of pathogenic bacteria in salsa. The aim of this study was to characterize the survival and growth of Salmonella, including the outbreak strain of Salmonella Saintpaul (E2003001236), in freshly made salsa and its main ingredients. Chopped tomatoes, jalapeño peppers, cilantro, and onions were tested individually or mixed according to different salsa recipes. Samples were inoculated with five Salmonella serotypes at 3 log CFU/g: Saintpaul (various strains), Typhimurium, Montevideo, Newport, or Enteritidis. Samples were then stored at room temperature (23°C) for up to 12 h or 3 days. The Salmonella Saintpaul levels reached approximately 9 log CFU/g after 2 days in tomato, jalapeño pepper, and cilantro. Growth was slower in onions, reaching 6 log CFU/g by day 3. Salsa recipes, with or without lime juice, supported the growth of Salmonella Saintpaul, and final levels were approximately 7 log CFU/g after 3 days at 23°C. In contrast, the counts of Salmonella Typhimurium, Salmonella Montevideo, Salmonella Newport, and Salmonella Enteritidis increased only 2 log CFU/g after 3 days in any of the salsas. Other Salmonella Saintpaul strains were able to grow in salsas containing 10% lime juice, but their final levels were less than 5 log CFU/g. These findings indicate the enhanced ability of the Salmonella Saintpaul outbreak strain to grow in salsa compared with other Salmonella strains. Recipe modifications including but not limited to adding lime juice (at least 10%) and keeping fresh salsa at room temperature for less than 12 h before consumption are strategies that can help mitigate the growth of Salmonella in salsa.

Melatonin regulates tumor aggressiveness under acidosis condition in breast cancer cell lines.

Breast cancer progression is composed of multiple steps that are influenced by tumor cell adaptations to survive under acidic conditions in the tumor microenvironment. Regulation of this cell survival behavior is a promising strategy to avoid cancer development. Melatonin is a natural hormone produced and secreted by the pineal gland capable of modulating different biological pathways in cancer. Although the anti-cancer effects of melatonin are currently widespread, its role in the acid tumor microenvironment remains poorly understood. The aim of the present study was to investigate the effect of low pH (6.7) on human breast cancer cell lines MCF-7 and MDA-MB-231, and the effectiveness of melatonin in acute acidosis survival mechanisms. Cell viability was measured by a MTT assay and the protein expression of glucose transporter (GLUT)-1, Ki-67 and caspase-3 was evaluated by immunocytochemical (ICC) analysis following low pH media and melatonin treatment. In both cell lines the viability was decreased after melatonin treatment (1 mM) under acidosis conditions for 24 h. ICC analysis showed a significant increase in GLUT-1 and Ki-67 expression at pH 6.7, and a decrease after treatment with melatonin for 12 and 24 h. The low pH media decreased the expression of caspase-3, which was increased after melatonin treatment for 12 and 24 h. Overall, the results of the present study revealed melatonin treatment increases apoptosis, as indicated by changes in caspase-3, and decreases proliferation, indicated by changes to Ki-67, and GLUT-1 protein expression under acute acidosis conditions in breast cancer cell lines.

Improvement of hydrogen yield of ethanol-producing Escherichia coli recombinants in acidic conditions

Background: An effective single culture with high glycerol consumption and hydrogen and ethanol coproduction yield is still in demand. A locally isolated glycerol-consuming Escherichia coli SS1 was found to produce lower hydrogen levels under optimized ethanol production conditions. Molecular approach was proposed to improve the hydrogen yield of E. coli SS1 while maintaining the ethanol yield, particularly in acidic conditions. Therefore, the effect of an additional copy of the native hydrogenase gene hycE and recombinant clostridial hydrogenase gene hydA on hydrogen production by E. coli SS1 at low pH was investigated. Results: Recombinant E. coli with an additional copy of hycE or clostridial hydA was used for fermentation using 10 g/L (108.7 mmol/L) of glycerol with an initial pH of 5.8. The recombinant E. coli with hycE and recombinant E. coli with hydA showed 41% and 20% higher hydrogen yield than wild-type SS1 (0.46 ± 0.01 mol/mol glycerol), respectively. The ethanol yield of recombinant E. coli with hycE (0.50 ± 0.02 mol/mol glycerol) was approximately 30% lower than that of wild-type SS1, whereas the ethanol yield of recombinant E. coli with hydA (0.68 ± 0.09 mol/mol glycerol) was comparable to that of wild-type SS1. Conclusions: Insertion of either hycE or hydA can improve the hydrogen yield with an initial pH of 5.8. The recombinant E. coli with hydA could retain ethanol yield despite high hydrogen production, suggesting that clostridial hydA has an advantage over the hycE gene in hydrogen and ethanol coproduction under acidic conditions. This study could serve as a useful guidance for the future development of an effective strain coproducing hydrogen and ethanol.

Root growth restraint can be an acclimatory response to low pH and is associated with reduced cell mortality: a possible role of class III peroxidases and NADPH oxidases.

Low pH (<5.0) can significantly decrease root growth but whether this is a direct effect of H(+) or an active plant response is examined here. Tomato (Solanum lycopersicum cv Micro-Tom) roots were exposed directly or gradually to low pH through step-wise changes in pH over periods ranging from 4 to 24 h. Roots exposed gradually to pH 4.5 grew even less than those exposed directly, indicating a plant-coordinated response. Direct exposure to pH 4.0 suppressed root growth and caused high cell mortality, in contrast to roots exposed gradually, in which growth remained inhibited but cell viability was maintained. Total class III peroxidase activity increased significantly in all low pH treatments, but was not correlated with the observed differential responses. Use of the enzyme inhibitors salicylhydroxamic acid (SHAM) or diphenyleneiodonium chloride (DPI) suggest that peroxidase and, to a lesser extent, NADPH oxidase were required to prevent or reduce injury in all low pH treatments. However, a role for other enzymes, such as the alternative oxidase is also possible. The results with SHAM, but not DPI, were confirmed in tobacco BY-2 cells. Our results indicate that root growth inhibition from low pH can be part of an active plant response, and suggest that peroxidases may have a critical early role in reducing loss of cell viability and in the observed root growth constraint.

Physiology of the fuel ethanol strain Saccharomyces cerevisiae PE-2 at low pH indicates a context-dependent performance relevant for industrial applications.

Selected Saccharomyces cerevisiae strains are used in Brazil to produce the hitherto most energetically efficient first-generation fuel ethanol. Although genome and some transcriptome data are available for some of these strains, quantitative physiological data are lacking. This study investigates the physiology of S. cerevisiae strain PE-2, widely used in the Brazilian fuel ethanol industry, in comparison with CEN.PK113-7D, a reference laboratory strain, focusing on tolerance to low pH and acetic acid stress. Both strains were grown in anaerobic bioreactors, operated as batch, chemostat or dynamic continuous cultures. Despite their different backgrounds, biomass and product formation by the two strains were similar under a range of conditions (pH 5 or pH < 3, with or without 105 mM acetic acid added). PE-2 displayed a remarkably higher fitness than CEN.PK113-7D during batch cultivation on complex Yeast extract - Peptone - Dextrose medium at low pH (2.7). Kinetics of viability loss of non-growing cells, incubated at pH 1.5, indicated a superior survival of glucose-depleted PE-2 cells, when compared with either CEN.PK113-7D or a commercial bakers' strain. These results indicate that the sulfuric acid washing step, used in the fuel ethanol industry to decrease bacterial contamination due to non-aseptic operation, might have exerted an important selective pressure on the microbial populations present in such environments.

An alternative method for evaluation of resistance to low pH and bile salts in probiotic characterization of Lactobacillus reuteri strains: a technical note

A considerable amount of attention has been focused on the use of lactobacilli as probiotics. The use of microorganisms as probiotics must meet certain criteria, among these; the resistance to low pH and bile salts is of great importance. Different procedures have been designed to evaluate tolerance to gastrointestinal transit. Since resistance to low pH and bile salts is critical in the evaluation of probiotic strains, the aim of this study was to compare resistance to bile salts for 20 strains of porcine Lactobacillus reuteri that had been isolated from healthy pigs, using two different procedures based on exposure to low pH. Statistical analysis revealed no difference in resistance of the strains to bile salts whether pH was low or not. The use of low pH prior to exposure to bile salts therefore more closely represents physiological conditions that should be used for the evaluation of potential porcine probiotic lactobacilli. Further studies using digestive enzymes are required to evaluate resistance to gastrointestinal transit, since it is an important factor in the evaluation of strains with potential probiotic activity.

Existe un interés creciente en el uso de lactobacilos como probióticos. El uso de microorganismos como probióticos, requiere que éstos cubran ciertos requisitos, dentro de los cuales la resistencia a pH ácido y a sales biliares, se consideran de suma importancia. Actualmente existen distintos procedimientos que permiten evaluar la tolerancia al tránsito gastrointestinal. Debido a que la resistencia a pH ácido y a sales biliares es una condición crítica dentro de la evaluación de cepas probióticas, la finalidad del presente trabajo consistió en comparar la resistencia a sales biliares de 20 cepas de Lactobacillus reuteri aisladas de cerdos sanos, mediante dos procedimientos diferentes basados en la exposición a pH ácido. El análisis estadístico no mostró diferencias en la resistencia de las cepas a sales biliares cuando se realizó una exposición previa a pH ácido. Así, la exposición a pH ácido previo a la incubación con sales biliares, asemeja más a las condiciones fisiológicas que el microorganismo encontrará de manera natural y es una buena alternativa en la evaluación probiótica de lactobacilos de origen porcino. Se requieren estudios que incluyan enzimas digestivas, con el fin de evaluar la resistencia al tránsito gastrointestinal, dado que es uno de los factores más importantes en la evaluación de cepas con potencial actividad probiótica.