Antimicrobial activity of extracts from macroalgae Ulva lactuca against clinically important Staphylococci is impacted by lunar phase of macroalgae harvest.

UNLABELLED: Staphylococcus aureus is a common human bacterial pathogen that causes skin and soft tissue infections. Methicillin-resistant Staph. aureus (MRSA) are increasingly drug-resistant, and thus there is great need for new therapeutics to treat Staph. aureus infections. Attention has focused on potential utility of natural products, such as extracts of marine macroalgae, as a source of novel antimicrobial compounds. The green macroalgae Ulva lactuca produces compounds inhibitory to human pathogens, although the effectiveness of U. lactuca extracts against clinically relevant strains of Staph. aureus is poorly understood. In addition, macroalgae produce secondary metabolites that may be influenced by exogenous factors including lunar phase, but whether lunar phase affects U. lactuca antimicrobial capacity is unknown. We sought to evaluate the antibacterial properties of U. lactuca extracts against medically important Staphylococci, and to determine the effect of lunar phase on antimicrobial activity. We report that U. lactuca methanolic extracts inhibit a range of Staphylococci, and that lunar phase of macrolagae harvest significantly impacts antimicrobial activity, suggesting that antimicrobial properties can be maximized by manipulating time of algal harvest. These findings provide useful parameters for future studies aimed at isolating and characterizing U. lactuca anti-Staphylococcal agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The growing prevalence of antibiotic-resistant human pathogens such as methicillin-resistant Staphylococcus aureus (MRSA) has intensified efforts towards discovery and development of novel therapeutics. Marine macroalgae like Ulva lactuca are increasingly recognized as potential sources of antimicrobials, but the efficacy of U. lactuca extracts against common, virulent strains of Staph. aureus is poorly understood. We demonstrate that U. lactuca methanolic extracts inhibit a variety of clinically relevant Staphylococcus strains, and that the antimicrobial activity can be maximized by optimizing time of algal harvest. These findings provide potentially useful parameters for future work of isolating and identifying novel antimicrobial agents from macroalgae.

Effect of sublethal heat stress on Salmonella Typhimurium virulence.

AIMS: To determine the virulence gene expression of Salmonella Typhimurium in response to sublethal heat stress and determine the adhesion and invasion pattern of heat-stressed Salmonella in Caco-2 intestinal epithelial cells. METHODS AND RESULTS: Transcriptional profiling was employed to capture the virulence gene response of Salm. Typhimurium at 42°C sublethal heat stress. Data indicated an induction of SPI-2 and SPI-5 genes and a repression of SPI-1-encoded genes due to heat stress. Gene expression pattern also showed induced transcription of fimbriae genes and genes present within the stress-associated Rpo regulon. Changes in adhesion and invasion pattern of heat-stressed Salm. Typhimurium were tested in Caco-2 cells. Heat-stressed Salm. Typhimurium showed greater adhesion to Caco-2 cells compared with nonstressed control cells. CONCLUSIONS: Salmonella Typhimurium exposed to sublethal heat stress responds by altered virulence gene expression, which further enhances the adhesion of bacterial cells to intestinal Caco-2 cells. Results indicate a role of physiological stress in Salm. Typhimurium in promoting microbial virulence and host cell vulnerability to infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Studying the Salmonella virulence genes expression in response to sublethal heat stress is crucial for the understanding of the virulence status of Salmonella in temperature-abused foods. Results of this study provide information about the gene response and virulence status of Salmonella pathogenicity factors in response to sublethal heat stress towards host cells.

Influence of stressors on normal intestinal microbiota, intestinal morphology, and susceptibility to Salmonella enteritidis colonization in broilers.

In modern poultry production systems, environmental stressors may influence bird performance and susceptibility to pathogens such as Salmonella Enteritidis. Experiments were conducted to determine the influence of 24-h feed withdrawal and 24-h exposure to high temperature (30 degrees C) on intestinal characteristics of broilers. Attachment of Salmonella Enteritidis to ileal tissue was determined using an in vitro ileal loop assay. Changes in commensal intestinal microbial populations were determined using denaturing gradient gel electrophoresis, and alterations in ileal morphology were determined histologically. Ex vivo attachment of Salmonella Enteritidis to ileal tissues increased by 1.5 logs (9.05 log10 vs. 7.59 log (10) Salmonella Enteritidis/g of ileal tissue; P = 0.0006) in broilers fasted for 24 h. Similarly, ileal tissues from birds subjected to 30 degrees C for 24 h had increased ex vivo attachment of Salmonella Enteritidis (8.77 log(10) vs. 8.50 log(10) Salmonella Enteritidis/g of ileum; P = 0.01) compared with birds held at 23 degrees C. Exposure to 30 degrees C for 24 h also altered microbial community structure in the ileum and cecum. Subjecting birds to 30 degrees C for 24 h reduced crypt depth (6.0 vs. 7.8 microm, respectively; P = 0.002), but had no effect on villus height or villus:crypt ratio. This research shows that acute stressors in poultry production systems can cause changes in the normal intestinal microbiota and epithelial structure, which may lead to increased attachment of Salmonella Enteritidis.

The effect of steam flaked or dry ground corn and supplemental phytic acid on nitrogen partitioning in lactating cows and ammonia emission from manure.

The effect of starch source and supplemental phytic acid (PA) on N partitioning and excretion and ammonia volatilization from dairy manure was evaluated with 8 midlactation cows. Cows were randomly assigned to treatments in replicated 4 x 4 Latin squares with four 18-d periods. Diets were 61% forage, 25% starch, 17.2% crude protein, and 31% neutral detergent fiber and included dry ground corn (DG) or steam flaked corn (SF) with no supplemental P (L; 0.34% P) or supplemental purified PA (0.45% P) to provide additional P from a non-mineral source. Total collection of milk, urine, and feces was conducted on d 16 to 18 of each period. Cows fed SF had lower dry matter (DM) intakes than those fed DG, which, in addition to increased starch digestibility and ruminal fermentation, contributed to higher DM digestibility. Cows fed SF had reduced feces and urine excretion compared with cows fed DG. Also, N intake for cows fed SF was lower, and N digestibility was higher, compared with cows fed DG; therefore, N excretion in both feces and urine was reduced in these cows. Despite the differences in DM intake, lactation performance was not affected by starch sources. Therefore, the efficiency of N utilization increased with SF. Addition of PA did not affect N intake or utilization. Feces and urine were subsampled from each cow, and wet feces and urine were mixed in sealed chambers in the proportions excreted. Ammonia volatilization was measured for 36 h using acid traps sampled on a planned time course. Nitrogen at time zero (A0), rate of ammonia emission (k), and residual N (R) were calculated using the exponential decay model At = A0 e(-kt) + R. Rate of ammonia loss from mixed feces and urine was lower from cows fed SF than from those fed DG. Altering dietary starch source to improve nutrient digestibility and to reduce N excretion by lactating cows may provide opportunity to reduce ammonia losses from manure.

Application of prebiotics and probiotics in poultry production.

The intestinal microbiota, epithelium, and immune system provide resistance to enteric pathogens. Recent data suggest that resistance is not solely due to the sum of the components, but that cross-talk between these components is also involved in modulating this resistance. Inhibition of pathogens by the intestinal microbiota has been called bacterial antagonism, bacterial interference, barrier effect, colonization resistance, and competitive exclusion. Mechanisms by which the indigenous intestinal bacteria inhibit pathogens include competition for colonization sites, competition for nutrients, production of toxic compounds, or stimulation of the immune system. These mechanisms are not mutually exclusive, and inhibition may comprise one, several, or all of these mechanisms. Consumption of fermented foods has been associated with improved health, and lactic acid bacteria (lactobacilli and bifidobacteria) have been implicated as the causative agents for this improved health. Research over the last century has shown that lactic acid bacteria and certain other microorganisms can increase resistance to disease and that lactic acid bacteria can be enriched in the intestinal tract by feeding specific carbohydrates. Increased bacterial resistance to antibiotics in humans has caused an increase in public and governmental interest in eliminating sub-therapeutic use of antibiotics in livestock. An alternative approach to sub-therapeutic antibiotics in livestock is the use of probiotic microorganisms, prebiotic substrates that enrich certain bacterial populations, or synbiotic combinations of prebiotics and probiotics. Research is focused on identifying beneficial bacterial strains and substrates along with the conditions under which they are effective.