Microbial Degradation of Lobster Shells to Extract Chitin Derivatives for Plant Disease Management.

Biodegradation of lobster shells by chitinolytic microorganisms are an environment safe approach to utilize lobster processing wastes for chitin derivation. In this study, we report degradation activities of two microbes, "S223" and "S224" isolated from soil samples that had the highest rate of deproteinization, demineralization and chitinolysis among ten microorganisms screened. Isolates S223 and S224 had 27.3 and 103.8 protease units mg-1 protein and 12.3 and 11.2 µg ml-1 of calcium in their samples, respectively, after 1 week of incubation with raw lobster shells. Further, S223 contained 23.8 µg ml-1 of N-Acetylglucosamine on day 3, while S224 had 27.3 µg ml-1 on day 7 of incubation with chitin. Morphological observations and 16S rDNA sequencing suggested both the isolates were Streptomyces. The culture conditions were optimized for efficient degradation of lobster shells and chitinase (∼30 kDa) was purified from crude extract by affinity chromatography. The digested lobster shell extracts induced disease resistance in Arabidopsis by induction of defense related genes (PR1 > 500-fold, PDF1.2 > 40-fold) upon Pseudomonas syringae and Botrytis cinerea infection. The study suggests that soil microbes aid in sustainable bioconversion of lobster shells and extraction of chitin derivatives that could be applied in plant protection.

Feed Supplementation with Red Seaweeds, Chondrus crispus and Sarcodiotheca gaudichaudii, Reduce Salmonella Enteritidis in Laying Hens.

Salmonella Enteritidis is vertically transmitted to eggs from laying hens through infected ovaries and oviducts. S. Enteritidis can also penetrate the eggshell from contaminated feces. Reducing S. Enteritidis in laying hens is vital to provide safer eggs and minimize the spread of salmonellosis to humans. Antibiotics have been widely used to control bacterial diseases in broilers and laying hens. However, there is a major concern that the use of antibiotics leads to the development of antibiotic resistance and adverse effects on microbiota of the treated birds. Thus, there is an interest in developing alternatives to antibiotics, such as dietary prebiotics. In the present study, feed supplemented with the red seaweeds: Chondrus crispus (CC) or Sarcodiotheca gaudichaudii (SG), was offered to laying hens late in production to control S. Enteritidis. Diets contained one of the following; 2% or 4% Chondrus crispus (CC2, and CC4, respectively) or Sarcodiotheca gaudichaudii (SG2 and SG4, respectively). Chlortetracycline was used in the positive control diet. During week-4, 48 birds were orally challenged with 2 × 109 CFU/mL of S. Enteritidis. Eggs and fecal samples were collected 1, 3, 5, and 7 days' post inoculation. Birds were euthanized and organs (ceca, ovary, liver, and spleen) were sampled and analyzed for the presence of S. Enteritidis, 7 days' post inoculation. Results showed that seaweed reduced the negative effect on body weight and egg production in S. Enteritidis-challenged laying hens. Analysis of fecal samples showed that the antibiotic (CTC) reduced S. Enteritidis in the intestinal tract and fecal samples, 3 days' post inoculation. Fecal samples from Chlortetracycline and CC4 supplemented birds tested negative for S. Enteritidis on days 5 and 7 post inoculation (lowest detection limit = 10-1). S. Enteritidis colonization in the ceca was also significantly reduced in birds fed CC (4%) and Chlortetracycline. Blood serum profiles revealed that there were no significant differences in serum aspartate aminotransferase (AST) and sodium. However, the level of serum immunoglobulin (IgA) was higher in the CC4 treatment. The relative abundance of Lactobacillus acidophilus was significantly higher in CC4 while, the abundance of the pathogenic bacteria, Clostridium perfringens and Salmonella Enteritidis were reduced compared to control. Results indicate that feed supplemented with 4% CC is effective in providing protection against Salmonella Enteritidis colonization in laying hens.

Red Seaweeds Sarcodiotheca gaudichaudii and Chondrus crispus down Regulate Virulence Factors of Salmonella Enteritidis and Induce Immune Responses in Caenorhabditis elegans.

Red seaweeds are a rich source of unique bioactive compounds and secondary metabolites that are known to improve human and animal health. S. Enteritidis is a broad range host pathogen, which contaminates chicken and poultry products that end into the human food chain. Worldwide, Salmonella outbreaks have become an important economic and public health concern. Moreover, the development of resistance in Salmonella serovars toward multiple drugs highlights the need for alternative control strategies. This study evaluated the antimicrobial property of red seaweeds extracts against Salmonella Enteritidis using the Caenorhabditis elegans infection model. Six red seaweed species were tested for their antimicrobial activity against S. Enteritidis and two, Sarcodiotheca gaudichaudii (SG) and Chondrus crispus (CC), were found to exhibit such properties. Spread plate assay revealed that SG and CC (1%, w/v) significantly reduced the growth of S. Enteritidis. Seaweed water extracts (SWE) of SG and CC, at concentrations from 0.4 to 2 mg/ml, significantly reduced the growth of S. Enteritidis (log CFU 4.5-5.3 and log 5.7-6.0, respectively). However, methanolic extracts of CC and SG did not affect the growth of S. Enteritidis. Addition of SWE (0.2 mg/ml, CC and SG) significantly decreased biofilm formation and reduced the motility of S. Enteritidis. Quantitative real-time PCR analyses showed that SWE (CC and SG) suppressed the expression of quorum sensing gene sdiA and of Salmonella Pathogenesis Island-1 (SPI-1) associated genes sipA and invF, indicating that SWE might reduce the invasion of S. Enteritidis in the host by attenuating virulence factors. Furthermore, CC and SG water extracts significantly improved the survival of infected C. elegans by impairing the ability of S. Enteritidis to colonize the digestive tract of the nematode and by enhancing the expression of C. elegans immune responsive genes. As the innate immune response pathways of C. elegans and mammals show a high degree of conservation, these results suggest that these SWE may also impart beneficial effects on animal and human health.

Performance of Seasonally and Continuously Loaded Constructed Wetlands Treating Dairy Farm Wastewater.

A 2-yr study compared the performance of seasonally and continuously loaded constructed wetlands treating dairy farm wastewater. One wetland was loaded during the growing season (GS) periods only, while the other was continuously loaded. Weekly samples were analyzed for 5-d biochemical oxygen demand (BOD), total suspended solids (TSS), total Kjeldahl N (TKN), total ammoniacal N (TAN), total P (TP), and . Annual average daily mass removal rates (kg ha) were similar for both wetlands in both years; however, seasonal differences were observed. With the exception of BOD in Year 2, average daily GS areal mass removal rates were higher for the seasonal wetland. However, GS mass exports from the seasonal wetland were higher by 28 to 94%, with the exception of BOD in Year 1. Annual mass reductions (MRs; %) for nutrients were higher for the continuous wetland in both years. Annual MRs were similar for in both years and for TSS in Year 2. Annual mass exports from the seasonal wetland were higher for nutrients and by 14 to 77% in both years. Pollutant MRs generally decreased during the nongrowing season (NGS) for the continuous wetland; however, in Year 2 when lower loading rates were used, the wetland still removed 84 to 99% of the pollutant masses. The continuous wetland also performed better during periods of high flow that occurred during the GS. Although there were minimal differences in annual treatment performance, continuously loaded systems require less additional infrastructure and should require less maintenance and may, therefore, be more attractive for agricultural applications.

Bacterial Pathogen Indicator Transport from Livestock Mortality Biopiles.

Biopiles can be used to dispose of slaughterhouse residuals (SLRs); however, the fate of pathogenic bacteria (e.g., pathogenic strains of , ) in these systems is not well understood. The transport of these bacteria in water leaching from the biopile could represent a significant contamination source. This research examined the transport of Enterobacteriaceae and Enterococcaceae indicator bacteria from SLR biopiles. Three biopiles (2.6 m wide by 4.6 m long by 1.8 m high) were formed on soil layers in concrete cells that allowed for real-time monitoring of environmental parameters, hydrologic flux, and indicator bacteria levels in effluent leaching from the piles. In biopile effluent, indicator bacteria populations decreased exponentially following biopile formation. Indicator bacteria loads in effluent constituted <0.01% of the initial indicator bacteria levels in the biopiles, which was attributed to retention, inactivation, and death. Nearly 90% of the total indicator bacteria loads coincided with large precipitation events (>15 mm d). Movement of the indicator bacteria through the biopiles and underlying soil appeared to be consistent with preferential flow phenomena. The populations of the Enterobacteriaceae indicators remained low in conditions of higher soil water content and lower biopile temperatures, whereas the Enterococcaceae indicator appeared to regrow in these conditions. This indicated that bacterial pathogen transport from a biopile could be a concern after the disappearance of conventional bacterial indicators, such as . Management considerations should attempt to divert excess water from entering a biopile, such as locating a biopile under a roof. Unsaturated biopile and soil conditions should be maintained to impede water flow through preferential pathways in the soil underneath a biopile.

Feed supplementation with red seaweeds, Chondrus crispus and Sarcodiotheca gaudichaudii, affects performance, egg quality, and gut microbiota of layer hens.

The aim of this study was to evaluate the effect of the inclusion of red seaweed supplementation to standard poultry diets on production performance, egg quality, intestinal histology, and cecal short-chain fatty acids in Lohmann Brown Classic laying hens. A total of 160 birds were randomly assigned to 8 treatment groups. Control hens were fed a basal layer diet; positive control hens were fed a diet containing 2% inulin; and 6 treatment groups were fed a diet containing one of the following; 0.5, 1, or 2% Chondrus crispus (CC0.5, CC1, and CC2, respectively) and one of the same 3 levels of Sarcodiotheca gaudichaudii (SG0.5, SG1, and SG2, respectively). Dietary supplementation had no significant effect on the feed intake, BW, egg production, fecal moisture content, and blood serum profile of the birds. The feed conversion ratio per gram of egg was significantly more efficient (P = 0.001) for CC2 and SG2 treatments. Moreover, SG1 supplementation increased egg yolk weight (P = 0.0035) and birds with CC1 supplementation had higher egg weight (P = 0.0006). The SG2 and CC2 groups had greater (P < 0.05) villus height and villus surface area compared with the control birds. Seaweed supplementation increased the abundance of beneficial bacteria [e.g., Bifidobacterium longum (4- to 14-fold), Streptococcus salivarius (4- to 15-fold)] and importantly reduced the prevalence of Clostridium perfringens in the gut of the chicken. Additionally, the concentrations of short-chain fatty acids, including acetic acid, propionic acid, n-butyric acid, and i-butyric acid, were significantly higher (P < 0.05) in CC and SG treatments than in the control. In conclusion, dietary supplementation using red seaweed inclusions can act as a potential prebiotic to improve performance, egg quality, and overall gut health in layer hens.

Wool-waste as organic nutrient source for container-grown plants.

A container experiment was conducted to test the hypothesis that uncomposted wool wastes could be used as nutrient source and growth medium constituent for container-grown plants. The treatments were: (1) rate of wool-waste application (0 or unamended control, 20, 40, 80, and 120 g of wool per 8-in. pot), (2) growth medium constituents [(2.1) wool plus perlite, (2.2) wool plus peat, and (2.3) wool plus peat plus perlite], and (3) plant species (basil and Swiss chard). A single addition of 20, 40, 80, or 120 g of wool-waste to Swiss chard (Beta vulgaris L.) and basil (Ocimum basilicum L.) in pots with growth medium provided four harvests of Swiss chard and five harvests of basil. Total basil yield from the five harvests was 1.6-5 times greater than the total yield from the unamended control, while total Swiss chard yield from the four harvests was 2-5 times greater relative to the respective unamended control. The addition of wool-waste to the growth medium increased Swiss chard and basil tissue N, and NO(3)-N and NH(4)-N in growth medium relative to the unamended control. Scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) microanalysis of wool fibers sampled at the end of the experiments indicated various levels of decomposition, with some fibers retaining their original surface structure. Furthermore, most of the wool fibers' surfaces contained significant concentrations of S and much less N, P, or K. SEM/EDX revealed that some plant roots grow directly on wool-waste fibers suggesting either (1) root directional growth towards sites with greater nutrient concentration and/or (2) a possible role for roots or root exudates in wool decomposition. Results from this study suggest that uncomposted wool wastes can be used as soil amendment, growth medium constituent, and nutrient source for container-grown plants.

Phosphorus losses through agricultural tile drainage in Nova Scotia, Canada.

Tile drainage water from agricultural fields commonly exceeds environmental guidelines for phosphorus (P) in rivers and streams. The loss of P through artificial drainage is spatially and temporally variable, and is related to local factors. This study characterizes variability in total P (TP) and soluble reactive P (SRP) concentrations in weekly drainage samples from 39 agricultural fields in Nova Scotia, Canada, from April 2002 through December 2003. We examined connections between P concentrations and the factors: (i) soil texture; (ii) discharge flow rate; (iii) soil test P (STP); (iv) manure type; and (v) crop cover. Generally, variability between fields and samples was great, and fields with standard deviations exceeding the mean for TP, SRP, and flow rate were 71, 54, and 79%, respectively. It was evident that poultry and swine manure contributed to high STPs, and to constantly high TP concentrations with high proportions of SRP. Concentrations varied from week to week, and particularly in April, May, October, and November when the greatest TP, SRP, and flow rate averages were measured. Mean TP concentrations exceed the USEPA (1994) TP guideline of 0.10 mg L(-1) at 82% of the fields, and periodically concentrations more than 10 times, and occasionally more than 50 times higher than the guideline were found. The proportion of SRP in TP had a tendency to be higher when TP levels were high in coarse textured soils. In Nova Scotia, dairy manure is most often applied on permanent cover crops, which did not show as much P concentration variability as crop rotations. Daily or hourly observation of short-term increases in P concentrations related to the described factors would help to characterize the changes in P concentrations observed during frequent heavy drainage flow events.