antiCD49d Ab treatment ameliorates age-associated inflammatory response and mitigates CD8+ T-cell cytotoxicity after traumatic brain injury.

Patients aged 65 years and older account for an increasing proportion of patients with traumatic brain injury (TBI). Older TBI patients experience increased morbidity and mortality compared to their younger counterparts. Our prior data demonstrated that by blocking α4 integrin, anti-CD49d antibody (aCD49d Ab) abrogates CD8+ T-cell infiltration into the injured brain, improves survival, and attenuates neurocognitive deficits. Here, we aimed to uncover how aCD49d Ab treatment alters local cellular responses in the aged mouse brain. Consequently, mice incur age-associated toxic cytokine and chemokine responses long-term post-TBI. aCD49d Ab attenuates this response along with a T helper (Th)1/Th17 immunological shift and remediation of overall CD8+ T cell cytotoxicity. Furthermore, aCD49d Ab reduces CD8+ T cells exhibiting higher effector status, leading to reduced clonal expansion in aged, but not young, mouse brains with chronic TBI. Together, aCD49d Ab is a promising therapeutic strategy for treating TBI in the older people. Graphic abstract: Aged brains after TBI comprise two pools of CD8 + T cells . The aged brain has long been resided by a population of CD8 + T cells that's exhaustive and dysfunctional. Post TBI, due to BBB impairment, functional CD8 + T cells primarily migrate into the brain parenchyma. Aged, injury-associated microglia with upregulated MHC class I molecules can present neoantigens such as neuronal and/or myelin debris in the injured brains to functional CD8+ T, resulting in downstream CD8+ T cell cytotoxicity. aCD49d Ab treatment exerts its function by blocking the migration of functional effector CD8 + T cell population, leading to less cytotoxicity and resulting in improved TBI outcomes in aged mice.

Podocyte-specific deletion of miR-146a increases podocyte injury and diabetic kidney disease.

Diabetic glomerular injury is a major complication of diabetes mellitus and is the leading cause of end stage renal disease (ESRD). Healthy podocytes are essential for glomerular function and health. Injury or loss of these cells results in increased proteinuria and kidney dysfunction and is a common finding in various glomerulopathies. Thus, mechanistic understanding of pathways that protect podocytes from damage are essential for development of future therapeutics. MicroRNA-146a (miR-146a) is a negative regulator of inflammation and is highly expressed in myeloid cells and podocytes. We previously reported that miR-146a levels are significantly reduced in the glomeruli of patients with diabetic nephropathy (DN). Here we report generation of mice with selective deletion of miR-146a in podocytes and use of these mice in models of glomerular injury. Induction of glomerular injury in C57BL/6 wildtype mice (WT) and podocyte-specific miR-146a knockout (Pod-miR146a-/-) animals via administration of low-dose lipopolysaccharide (LPS) or nephrotoxic serum (NTS) resulted in increased proteinuria in the knockout mice, suggesting that podocyte-expressed miR-146a protects these cells, and thus glomeruli, from damage. Furthermore, induction of hyperglycemia using streptozotocin (STZ) also resulted in an accelerated development of glomerulopathy and a rapid increase in proteinuria in the knockout animals, as compared to the WT animals, further confirming the protective role of podocyte-expressed miR-146a. We also confirmed that the direct miR-146a target, ErbB4, was significantly upregulated in the diseased glomeruli and erlotinib, an ErbB4 and EGFR inhibitor, reducedits upregulation and the proteinuria in treated animals. Primary miR146-/- podocytes from these animals also showed a basally upregulated TGFß-Smad3 signaling in vitro. Taken together, this study shows that podocyte-specific miR-146a is imperative for protecting podocytes from glomerular damage, via modulation of ErbB4/EGFR, TGFß, and linked downstream signaling.

CD11b agonists offer a novel approach for treating lupus nephritis.

Lupus nephritis (LN) develops in more than a third of all systemic lupus erythematosus (SLE) patients and is the strongest predictor of morbidity and mortality. Increased circulating levels of type I interferon (IFN I) and anti-double stranded DNA (anti-dsDNA) and anti-RNA binding protein (anti-RNP) antibodies lead to increased glomerular injury via leukocyte activation and glomerular infiltration. Uncontrolled Toll-like receptor (TLR) signaling in leukocytes results in increased production of IFN I and anti-dsDNA antibodies. ITGAM gene codes for integrin CD11b, the α-chain of integrin heterodimer CD11b/CD18, that is highly expressed in leukocytes and modulates TLR-dependent pro-inflammatory signaling. Three nonsynonymous SNPs in the ITGAM gene strongly correlate with increased risk for SLE and LN and with IFN I levels. Here we review the literature on the role of CD11b on leukocytes in LN. We also incorporate conclusions from several recent studies that show that these ITGAM SNPs result in a CD11b protein that is less able to suppress TLR-dependent pro-inflammatory pathways in leukocytes, that activation of CD11b via novel small molecule agonists suppresses TLR-dependent pathways, including reductions in circulating levels of IFN I and anti-dsDNA antibodies, and that CD11b activation reduces LN in model systems. Recent data strongly suggest that integrin CD11b is an exciting new therapeutic target in SLE and LN and that allosteric activation of CD11b is a novel therapeutic paradigm for effectively treating such autoimmune diseases.

Impacts of volcanic eruptions and early recovery in freshwater environments and organisms.

Volcanic eruptions modify environments physically and chemically with serious consequences for the biota. In this review, we analysed 80 papers reporting the effects of volcanic eruptions in freshwater environments and on freshwater organisms. An increase in water turbidity is the most common reported physical effect while increases in concentrations of inorganic elements, many representing nutrients for primary producers, are the most common chemical effects. Bacterial growth is usually stimulated, while autotrophs can be either positively or negatively affected depending on the type of impact. A persistent effect reported in the biota is changes to the assemblage, which could generate further changes in terms of ecosystem functions. This analysis also identifies some information gaps, particularly involving the effects of eruptions on heterotrophic biofilms in streams and on invertebrates and fish in lakes. Most studies were carried out soon after the volcanic eruption, so it is difficult to assess the recovery of the ecosystems. Eruptions present unique opportunities for scientific discovery, although such studies are often hindered by a lack of pre-eruption data, which would allow for a more comprehensive assessment of the effects.

[Impact of confinement measures on the Covid-19 pandemic on alcohol risk consumption.] / Impacto de las medidas de confinamiento durante la pandemia de Covid-19 en el consumo de riesgo de alcohol.

OBJECTIVE: The public health strategies adopted for the containment of COVID-19 have generated psychosocial stressors that act as risk factors for alcohol consumption. The objectives of this study were to establish whether alcohol risk consumption varied during COVID-19 confinement, and how these variations manifested as a function of different sociodemographic variables. METHODS: The study was conducted during confinement with 3,779 participants in Spain. We used an online survey with sociodemographic variables (sex, age and employment situation) and AUDIT-C. Frequency and mean difference analysis were performed (Student t, ANOVA and Kruskal-Wallis) to establish intra-group variations before the pandemic compared to during confinement, and intergroup for each time period, reporting effect size. RESULTS: Alcohol risky consumption presented high prevalence rates before confinement (25,9% of alcohol consumers), showing a general decrease during confinement (15,1%), both in men (X2=224,829; p<0,001) and women (X2=214,324; p<0,001). Women had higher risk consumption rates both before the pandemic (X2=13,124; p<0,001; d=1,067) and during confinement (X2=26,316; p<0,001; d=2,85); however, men reported higher score in AUDIT-C before the pandemic (t(2343)=-7,887; p<0,001; d=0,322) and during confinement (t(2343)=-5,664; p<0,001; d=0,231). Considering age, significant differences in prevalence among groups were found before the pandemic (X2=22,889; p<0,001) and during confinement (X2=38,302; p<0,001), with the age range 18-24 showing less prevalence. Differences among age groups were also found in the risky consumption scores during confinement (F(5,2338)=43,849; p=0,001), increasing with age. With regards to the employment situation, differences before the pandemic (KW(5,2933)=13,467; p=0,019; E2R=0,44), and during confinement (KW(5,2933)=149,818; p<0,001; E2R=0,51) were also found. Self-employed workers showed a higher score in alcohol risky consumption with respect to full-time employees (p=0,047). CONCLUSIONS: The alcohol risky consumption decreased in a generalized way during the confinement by COVID-19, but the changes in prevalence and risk index were dependent on sociodemographic variables. It is recommended that these findings are considered in the design of public health policies and strategies.

OBJETIVO: Las estrategias de salud pública adoptadas para la contención del COVID-19 han generado estresores psicosociales que actúan como factores de riesgo del consumo de alcohol. Los objetivos del estudio fueron establecer si el consumo de riesgo de alcohol varió durante el confinamiento por COVID-19 durante los meses de abril y mayo de 2020, y cómo estas variaciones se manifestaban en función de distintas variables sociodemográficas. METODOS: El estudio se desarrolló durante el confinamiento con 3.779 participantes en España. Se utilizó una encuesta online con variables sociodemográficas (sexo, edad y situación laboral) y el AUDIT-C. Se realizó análisis de frecuencias y diferencia de medias (t de Student, ANOVA y Kruskal-Wallis) para establecer las variaciones a nivel intragrupo entre antes de la pandemia y durante el confinamiento, e intergrupo para cada periodo temporal, reportando tamaño del efecto. RESULTADOS: El consumo de riesgo de alcohol presentó una prevalencia elevada antes del confinamiento (25,9% de los consumidores de alcohol), mostrando un descenso durante el confinamiento (15,1%), tanto en hombres (X2=224,829; p<0,001) como en mujeres (X2=214,324; p<0,001). Las mujeres presentaron una mayor prevalencia de consumo de riesgo antes de la pandemia (X2=13,124; p<0,001; d=1,067) como durante el confinamiento (X2=26,316; p<0,001; d=2,85); sin embargo, los hombres reportaron mayores puntuaciones que las mujeres en el AUDIT-C antes de la pandemia (t(2343)=-7,887; p<0,001; d=0,322) y durante el confinamiento (t(2343)=-5,664; p<0,001; d=0,231). A nivel de edad, se encontraron diferencias significativas de prevalencia entre los distintos grupos antes de la pandemia (X2=22,889; p<0,001) y durante el confinamiento (X2=38,302; p<0,001), siendo el rango de 18-24 años el de menor prevalencia. También se observaron diferencias entre los grupos de edad en las puntuaciones de consumo de riesgo durante el confinamiento (F(5,2338)=43,849; p=0,001), aumentando conforme es mayor la edad. Respecto a la situación laboral, existen diferencias tanto antes de la pandemia (KW(5,2933)=13,467; p=0,019; E2R=0,44), como durante el confinamiento (KW(5,2933)=149,818; p<0,001; E2R=0,51). Ser trabajador autónomo mostró una mayor puntuación de consumo de riesgo de alcohol con respecto a ser trabajador a tiempo completo (p=0,047). CONCLUSIONES: El consumo de riesgo de alcohol desciende de manera generalizada durante el confinamiento por COVID-19, pero los cambios en prevalencia y probabilidad de ser consumidor de riesgo de alcohol son dependientes de variables sociodemográficas. Se recomienda contemplar estos hallazgos en el diseño de políticas y estrategias de salud pública.

Polyunsaturated Phospholipid Modified Membrane Degradation Catalyzed by a Secreted Phospholipase A2.

To optimize the compositions of the lipid-based nanomedicine and to advance understanding of the roles of polyunsaturated phospholipids in biological membranes, this study examined the effects of polyunsaturated phospholipids on the degradation of giant unilamellar vesicles catalyzed by a secreted phospholipase A2 (sPLA2) using fluorescence microscopy. Molecular interfacial packing, interaction, and degradation of the films containing various mixing ratios of saturated and polyunsaturated phospholipids were quantified using a Langmuir trough integrated with synchrotron X-ray surface scattering techniques. It was found that a high molar fraction (0.63 and above) of polyunsaturated phospholipids not only enhanced the rate of sPLA2-catalyzed vesicle degradation but also changed the vesicle deformation process and degradation product morphology. Hydrolysis of the saturated phospholipids generated highly ordered liquid crystal domains, which was reduced or prohibited by the presence of the polyunsaturated phospholipids in the reactant film.

Nutritional stress by means of high C:N ratios in the diet and starvation affects nitrogen isotope ratios and trophic fractionation of omnivorous copepods.

Nutritional stress, from feeding on low-quality diets or starvation, may cause changes in consumers' nitrogen isotope ratios (δ15N = 15N/14N) and trophic fractionation (∆15N = δ15Nconsumer - δ15Nfood source), however, research has shown mixed results in the magnitude and the direction of the change. This is potentially more complex for omnivores whose diets span a wide range of food resources. We conducted seasonal field samplings in Patagonian lakes and analyzed the relationship between seston (SES) quality parameters and the δ15N and ∆15N of an omnivorous copepod, Boeckella gracilipes (Bg). We also performed a 7-day laboratory starvation experiment, an extreme form of nutritional stress, to investigate if lack of food led to changes in δ15NBg values. Our field results showed that increasing values of the seston carbon to nitrogen ratio (C:NSES), chlorophyll a (Chl a), and δ15NSES were related to higher δ15NBg values. C:NSES and Chl a were also positively related to ∆15N; yet, C:NSES alone explained 70% of the variation. C:NSES values correlated with the presence of mixotrophic algae and ciliates that are key food resources for B. gracilipes. In our laboratory starvation experiment, the δ15NBg values increased significantly, pointing to use of internal N sources; yet, the change associated with starvation was less pronounced than that related to C:NSES changes in the field, suggesting depletion of the substrate pool in the former. We found that ∆15N values of omnivorous species consuming a low-quality diet would be higher than that from a conspecific with a high-quality diet; though fasting animals would show intermediate values.

Molecular interactions of phospholipid monolayers with a model phospholipase.

The intrinsic overexpression of secretory phospholipase A2 (sPLA2) in various pro-inflammatory diseases and cancers has the potential to be exploited as a therapeutic strategy for diagnostics and treatment. To explore this potential and advance our knowledge of the role of sPLA2 in related diseases, it is necessary to systematically investigate the molecular interaction of the enzyme with lipids. By employing a Langmuir trough integrated with X-ray reflectivity and grazing incidence X-ray diffraction techniques, this study examined the molecular packing structure of 1,2-palmitoyl-sn-glycero-3-phosphocholine (DPPC) films before and after enzyme adsorption and enzyme-catalyzed degradation. Molecular interaction of sPLA2 (from bee venom) with the DPPC monolayer exhibited Ca2+ dependence. DPPC molecules at the interface without Ca2+ retained a monolayer organization; upon adsorption of sPLA2 to the monolayer the packing became tighter. In contrast, sPLA2-catalyzed degradation of DPPC occurred in the presence of Ca2+, leading to disruption of the ordered monolayer structure of DPPC. The interfacial film became a mixture of highly ordered multilayer domains of palmitic acid (PA) and loosely packed monolayer phase of 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (lysoPC) that potentially contained the remaining un-degraded DPPC. The redistribution of lipid degradation products into the third dimension, which produced multilayer PA domains, damaged the structural integrity of the original lipid layer and may explain the bursting of liposomes observed in other studies after a latency period of mixing liposomes with sPLA2. A quantitative understanding of the lipid packing and lipid-enzyme interaction provides an intuitive means of designing and optimizing lipid-related drug delivery systems.

Global patterns and drivers of ecosystem functioning in rivers and riparian zones.

River ecosystems receive and process vast quantities of terrestrial organic carbon, the fate of which depends strongly on microbial activity. Variation in and controls of processing rates, however, are poorly characterized at the global scale. In response, we used a peer-sourced research network and a highly standardized carbon processing assay to conduct a global-scale field experiment in greater than 1000 river and riparian sites. We found that Earth's biomes have distinct carbon processing signatures. Slow processing is evident across latitudes, whereas rapid rates are restricted to lower latitudes. Both the mean rate and variability decline with latitude, suggesting temperature constraints toward the poles and greater roles for other environmental drivers (e.g., nutrient loading) toward the equator. These results and data set the stage for unprecedented "next-generation biomonitoring" by establishing baselines to help quantify environmental impacts to the functioning of ecosystems at a global scale.

Sustained effects of volcanic ash on biofilm stoichiometry, enzyme activity and community composition in North- Patagonia streams.

Volcanic eruptions are extreme perturbations that affect ecosystems. These events can also produce persistent effects in the environment for several years after the eruption, with increased concentrations of suspended particles and the introduction of elements in the water column. On 4th June 2011, the Puyehue-Cordón Caulle Volcanic Complex (40.59°S-72.11°W, 2200m.a.s.l.) erupted explosively in southern Chile. The area affected by the volcano was devastated; a thick layer of volcanic ash (up to 30cm) was deposited in areas 50 km east of the volcano towards Argentina. The aim of the present study was to evaluate the effect of volcanic ash deposits on stream ecosystems four years after the eruption, comparing biofilm stoichiometry, alkaline phosphatase activity, and primary producer's assemblage in streams which were severely affected by the volcano with unaffected streams. We confirmed in the laboratory that ash deposited in the catchment of affected streams still leach phosphorus (P) into the water four years after eruption. Results indicate that affected streams still receive volcanic particles and that these particles release P, thus stream water exhibits high P concentration. Biofilm P content was higher and the C:P ratio lower in affected streams compared to unaffected streams. As a consequence of less P in unaffected streams, the alkaline phosphatase activity was higher compared to affected streams. Cyanobacteria increased their abundances (99.9% of total algal biovolume) in the affected streams suggesting that the increase in P may positively affect this group. On the contrary, unaffected streams contained a diatom dominant biofilm. In this way, local heterogeneity was created between sub-catchments located within 30 km of each other. These types of events should be seen as opportunities to gather valuable ecological information about how severe disturbances, like volcanic eruptions, shape landscapes and lotic systems for several years after the event.

Shifts in microbial community structure and function in light- and dark-grown biofilms driven by warming.

Biofilms are dynamic players in biogeochemical cycling in running waters and are subjected to environmental stressors like those provoked by climate change. We investigated whether a 2°C increase in flowing water would affect prokaryotic community composition and heterotrophic metabolic activities of biofilms grown under light or dark conditions. Neither light nor temperature treatments were relevant for selecting a specific bacterial community at initial phases (7-day-old biofilms), but both variables affected the composition and function of mature biofilms (28-day-old). In dark-grown biofilms, changes in the prokaryotic community composition due to warming were mainly related to rotifer grazing, but no significant changes were observed in functional fingerprints. In light-grown biofilms, warming also affected protozoan densities, but its effect on prokaryotic density and composition was less evident. In contrast, heterotrophic metabolic activities in light-grown biofilms under warming showed a decrease in the functional diversity towards a specialized use of several carbohydrates. Results suggest that prokaryotes are functionally redundant in dark biofilms but functionally plastic in light biofilms. The more complex and self-serving light-grown biofilm determines a more buffered response to temperature than dark-grown biofilms. Despite the moderate increase in temperature of only 2°C, warming conditions drive significant changes in freshwater biofilms, which responded by finely tuning a complex network of interactions among microbial populations within the biofilm matrix.

Effect of a nutritional reconditioning program for thin dairy cattle on body weight, carcass quality, and fecal pathogen shedding.

OBJECTIVE: To assess changes in body weight, carcass quality, and fecal pathogen shedding in cull dairy cows fed a high-energy ration for 28 or 56 days prior to slaughter. DESIGN: Randomized clinical trial. ANIMALS: 31 adult Holstein dairy cows. PROCEDURES: Cows were randomly assigned to a control (immediate slaughter) group or a 28-day or 56-day feeding group. Cows in the feeding groups received a high-energy feed and were weighed every 7 days. Carcasses were evaluated by USDA employees. Fecal and blood samples were collected at the start and end of the feeding periods. RESULTS: Body condition score and adjusted preliminary yield grade were significantly increased in both feeding groups, compared with values for the control group; body weight, hot carcass weight, dressing percentage, and ribeye area were significantly increased after 56 days, but not after 28 days, compared with values for the control group. Average daily gain and marbling score were significantly lower after feeding for 28 days versus after 56 days. Prevalence of Escherichia coli O157:H7 shedding in feces decreased from 14% to 5.6%, but this difference was not significant. Cows seropositive for antibodies against bovine leukemia virus that had signs of lymphoma and lame cows had a low average daily gain. Net loss was $71.32/cow and $112.80/cow for the 28-day and 56-day feeding groups, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Feeding market dairy cows improved body condition and carcass quality. Cows seropositive for antibodies against bovine leukemia virus that have signs of lymphoma and lame cows might be poor candidates for reconditioning.

Prevalence and molecular characterization of Escherichia coli O157:H7 by multiple-locus variable-number tandem repeat analysis and pulsed-field gel electrophoresis in three sheep farming operations in California.

A year-long study was conducted to determine the fecal prevalence of Escherichia coli O157:H7 in three sheep ranches. Strain diversity and persistence were compared with multiple-locus variable-number tandem repeat analysis and pulsed-field gel electrophoresis. Ranch C, a feedlot, consisted of young sheep raised predominantly on a high-grain diet. The other two sites consisted of sheep raised on native pasture and a combination of native and irrigated pasture. Forty fecal samples were collected every month from each ranch. Samples were examined for E. coli O157:H7 by immunomagnetic separation and culture of the magnetic beads onto selective media. Detection of virulence markers in positive isolates was determined by PCR. E. coli O157:H7 was isolated from 100 (22.7%) of 440 fecal samples collected from ranch C. On ranch B, 9 (1.9%) of the 480 fecal samples were positive for the pathogen, while none of the samples from ranch A were positive. On ranch C, the odds of detecting E. coli O157:H7 was 3.2 times greater during the warmer months compared with the cooler months of the year. There was no association between days spent in the feedlot and fecal prevalence of the pathogen (P = 0.62). Most multiple-locus variable-number tandem repeat analysis types were isolated only once from ranch C (14 of 23), but several strains were isolated over 4 to 6 months, often in many intervening negative months. This study revealed that the prevalence of E. coli O157:H7 can be high in some sheep ranches in California, especially in feedlots where young sheep are fed predominantly high-grain rations.

Global distribution of a key trophic guild contrasts with common latitudinal diversity patterns.

Most hypotheses explaining the general gradient of higher diversity toward the equator are implicit or explicit about greater species packing in the tropics. However, global patterns of diversity within guilds, including trophic guilds (i.e., groups of organisms that use similar food resources), are poorly known. We explored global diversity patterns of a key trophic guild in stream ecosystems, the detritivore shredders. This was motivated by the fundamental ecological role of shredders as decomposers of leaf litter and by some records pointing to low shredder diversity and abundance in the tropics, which contrasts with diversity patterns of most major taxa for which broad-scale latitudinal patterns haven been examined. Given this evidence, we hypothesized that shredders are more abundant and diverse in temperate than in tropical streams, and that this pattern is related to the higher temperatures and lower availability of high-quality leaf litter in the tropics. Our comprehensive global survey (129 stream sites from 14 regions on six continents) corroborated the expected latitudinal pattern and showed that shredder distribution (abundance, diversity and assemblage composition) was explained by a combination of factors, including water temperature (some taxa were restricted to cool waters) and biogeography (some taxa were more diverse in particular biogeographic realms). In contrast to our hypothesis, shredder diversity was unrelated to leaf toughness, but it was inversely related to litter diversity. Our findings markedly contrast with global trends of diversity for most taxa, and with the general rule of higher consumer diversity at higher levels of resource diversity. Moreover, they highlight the emerging role of temperature in understanding global patterns of diversity, which is of great relevance in the face of projected global warming.

A global experiment suggests climate warming will not accelerate litter decomposition in streams but might reduce carbon sequestration.

The decomposition of plant litter is one of the most important ecosystem processes in the biosphere and is particularly sensitive to climate warming. Aquatic ecosystems are well suited to studying warming effects on decomposition because the otherwise confounding influence of moisture is constant. By using a latitudinal temperature gradient in an unprecedented global experiment in streams, we found that climate warming will likely hasten microbial litter decomposition and produce an equivalent decline in detritivore-mediated decomposition rates. As a result, overall decomposition rates should remain unchanged. Nevertheless, the process would be profoundly altered, because the shift in importance from detritivores to microbes in warm climates would likely increase CO(2) production and decrease the generation and sequestration of recalcitrant organic particles. In view of recent estimates showing that inland waters are a significant component of the global carbon cycle, this implies consequences for global biogeochemistry and a possible positive climate feedback.

Biofilm formation at warming temperature: acceleration of microbial colonization and microbial interactive effects.

River biofilms that grow on wet benthic surface are mainly composed of bacteria, algae, cyanobacteria and protozoa embedded in a polysaccharide matrix. The effects of increased river water temperature on biofilm formation were investigated. A laboratory experiment was designed employing two temperatures (11.1-13.2°C, night-day; 14.7-16.0°C, night-day) and two nutrient levels (0.054 mg P l(-1), 0.75 mg N l(-1); 0.54 mg P l(-1), 7.5 mg N l(-1)). Biofilm formation at the higher temperature was faster, while the biomass of the mature biofilm was mainly determined by nutrient availability. The specific response of the three microbial groups that colonized the substrata (algae, bacteria and ciliates) was modulated by interactions between them. The greater bacterial growth rate and earlier bacterial colonization at the higher temperature and higher nutrient status was not translated into the accrual of higher bacterial biomass. This may result from ciliates grazing on the bacteria, as shown by an earlier increase in peritrichia at higher temperatures, and especially at high nutrient conditions. Temperature and ciliate grazing might determine the growth of a distinctive bacterial community under warming conditions. Warmer conditions also produced a thicker biofilm, while functional responses were much less evident (increases in the heterotrophic utilization of polysaccharides and peptides, but no increase in primary production and respiration). Increasing the temperature of river water might lead to faster biofilm recolonization after disturbances, with a distinct biofilm community structure that might affect the trophic web. Warming effects would be expected to be more relevant under eutrophic conditions.

Longitudinal prevalence and molecular typing of Escherichia coli O157:H7 by use of multiple-locus variable-number tandem-repeat analysis and pulsed-field gel electrophoresis in fecal samples collected from a range-based herd of beef cattle in California.

OBJECTIVE: To evaluate seasonal patterns and risk factors for Escherichia coli O157:H7 in feces in a beef cattle herd and determine strain diversity and transition in E coli over time by use of multiple-locus variable-number tandem-repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE). SAMPLE POPULATION: 456 samples of freshly passed feces collected over a 1-year period from cattle in a range-based cow-calf operation located in the foothills of the Sierra Nevada Mountains in California. PROCEDURES: E coli O157:H7 was recovered from feces by use of immunomagnetic separation and 2 selective media. Virulence factors were detected via reverse transcriptase-PCR assay. Escherichia coli O157:H7 isolates were subtyped with MLVA and PFGE. Prevalence estimates were calculated and significant risk factors determined. A dendrogram was constructed on the basis of results of MLVA typing. RESULTS: Overall prevalence estimate for E coli O157:H7 was 10.5%, with the prevalence lowest during the winter. Mean temperature during the 30 days before collection of samples was significantly associated with prevalence of E coli O157:H7 in feces. Nineteen MLVA and 12 PFGE types were identified. CONCLUSIONS AND CLINICAL RELEVANCE: A seasonal pattern was detected for prevalence of E coli O157:H7 in feces collected from beef cattle in California. Subtyping via MLVA and PFGE revealed a diversity of E coli O157:H7 strains in a cow-calf operation and noteworthy turnover of predominant types. Given the importance of accurately determining sources of contamination in investigations of disease outbreaks in humans, MLVA combined with PFGE should be powerful tools for epidemiologists.

Plantas medicinales: Efecto antibacteriano in vitro de Plantago major L, Erythroxylum novogranatense, Plowman var truxillense y Camellia sinensis sobre bacterias de importancia estomatológica / Medicinal plants: Antibacterial effect in vitro de Plantago major L, Erythroxylum novogranatense, Plowman var truxillense and Camellia sinensis on stomatologic importance bacteria

El objetivo de la investigación fue comparar la actividad antibacteriana in vitro de los extractos hidroalcohólicos de tres plantas medicinales: Plantago major L. (llantén), Erythroxylum novograntense var truxillense (coca trujillo) y Camellia sinensis (té verde) mediante el método de difusión en agar con discos, sobre cinco cepas patrones de bacterias orales: Streptococcus mutans ATCC 25175, Lactobacillus acidophilus. ATCC 314, Actinomyces viscosus ATCC 15987, Prevotella melaninogenicus ATCC 25845 y Fusobacterium nucleatum ATCC 25586. Se obtuvieron extractos hidroalcohólicos de principios activos totales procedentes de las hojas secas de cada una de las plantas, mediante maceración alcohólica con alcohol etílico al 70 % y posterior evaporación del solvente con el empleo del rotavapor. Cada extracto se diluyó en alcohol etílico al 70 % en las concentraciones de 25 µg/mL y 50 µg/mL. Estas soluciones fueron comparadas con PerioAid® (clorhexidina 0,012 %) como control positivo y con alcohol etílico al 70 %, como control negativo. Al realizar las pruebas de sensibilidad in vitro, se obtuvieron los siguientes resultados: los tres extractos hidroalcohólicos en ambas concentraciones presentaron actividad antibacteriana mayor al alcohol etílico (5,8 mm) y menor que el PerioAid® (22,0 mm) sobre las cinco cepas bacterianas en estudio. La mayor actividad presentó el extracto hidroalcohólico de Camellia sinensis a 50 µg/mL, la menor actividad presento Plantago major a 25 µg/mL. Se concluye que los tres extractos hidroalcohólicos en las diluciones de 25 y 50 µg/mL presentaron actividad antibacteriana sobre Streptococcus mutans, Lactobacillus acidophilus, Actinomyces viscosus, Prevotella melaninogenica y Fusobacterium nucleatum. El efecto antibacteriano aumentó con la concentración en P. melaninogenica, que fue la cepa más sensible y A. viscosus la menos sensible.