High-Throughput Measurement and Classification of Organic P in Environmental Samples

Many types of organic phosphorus (P) molecules exist in environmental samples¹. Traditional P measurements do not detect these organic P compounds since they do not react with colorimetric reagents^2,3. Enzymatic hydrolysis (EH) is an emerging method for accurately characterizing organic P forms in environmental samples^4,5. This method is only trumped in accuracy by Phosphorus-31 Nuclear Magnetic Resonance Spectroscopy (³¹P-NMR) -a method that is expensive and requires specialized technical training⁶. We have adapted an enzymatic hydrolysis method capable of measuring three classes of phosphorus (monoester P, diester P and inorganic P) to a microplate reader system⁷. This method provides researchers with a fast, accurate, affordable and user-friendly means to measure P species in soils, sediments, manures and, if concentrated, aquatic samples. This is the only high-throughput method for measuring the forms and enzyme-lability of organic P that can be performed in a standard laboratory. The resulting data provides insight to scientists studying system nutrient content and eutrophication potential.     

Dermal excisional wound healing in pigs following treatment with topically applied pure oxygen

Hypoxia, caused by disrupted vasculature and peripheral vasculopathies, is a key factor that limits dermal wound healing. Factors that can increase oxygen delivery to the regional tissue, such as supplemental oxygen, warmth, and sympathetic blockade, can accelerate healing. Clinical experience with adjunctive hyperbaric oxygen therapy (HBOT) in the treatment of chronic wounds have shown that wound hyperoxia may increase granulation tissue formation and accelerate wound contraction and secondary closure. However, HBOT is not applicable to all wound patients and may pose the risk of oxygen toxicity. Thus, the efficacy of topical oxygen treatment in an experimental setting using the pre-clinical model involving excisional dermal wound in pigs was assessed. Exposure of open dermal wounds to topical oxygen treatment increased tissue pO₂ of superficial wound tissue. Repeated treatment accelerated wound closure. Histological studies revealed that the wounds benefited from the treatment. The oxygen treated wounds showed signs of improved angiogenesis and tissue oxygenation. Topically applied pure oxygen has the potential of benefiting some wound types. Further studies testing the potential of topical oxygen in pre-clinical and clinical settings are warranted.     

Declines of zoonotic agents in liquid livestock wastes stored in batches on-farm

AIM: To measure the decline rates of zoonotic agents introduced into liquid livestock wastes in on-farm storage tanks. METHODS AND RESULTS: Salmonella spp., Escherichia coli O157, Campylobacter jejuni, Listeria monocytogenes and Cryptosporidium parvum, propagated in laboratory-controlled conditions, were inoculated into 35,000-l volumes of fresh livestock wastes (pig slurries, cattle slurries and dirty waters). D-values for bacteria were six to 44 days, and for C. parvum were 133 to 345 days. Campylobacter jejuni declined significantly more rapidly than the other bacterial pathogens, while E. coli O157 declined significantly more slowly. On average, bacterial declines were not affected by the season of waste deposition and storage or by the dry matter content of the wastes, but were more rapid in dirty waters than in pig slurries. The physiciochemical composition of wastes in each category varied significantly. CONCLUSIONS: Zoonotic agents can survive for several months during storage of liquid livestock wastes. Livestock wastes should be batch-stored and not subjected to continuous additions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that batches of liquid livestock waste, if contaminated with bacterial pathogens, should be stored for 6 months to reduce contamination levels. Alternative strategies for reducing C. parvum levels in liquid livestock wastes should be explored.     

A survey of ammonia-assimilating micro-organisms in cattle manure composting

AIMS: To evaluate the ammonia-assimilating abilities of micro-organisms isolated from cattle manure composting processes and to determine the distribution of cultivable species of ammonia-assimilating micro-organisms in microbial communities during the composting processes. METHODS AND RESULTS: Compost samples were collected from four stages of treatment. Trypto soya agar was used for the isolation of ammonia-assimilating aerobes. Many of the isolates showed high ammonia-assimilating ability in a medium containing basal components and a compost extract. Partial 16S ribosomal DNA sequencing showed that the cultivable species of highly efficient ammonia-assimilating isolates changed during the composting process. The community structure of micro-organisms and actinomycetes was analysed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Two species of actinomycetes identified by PCR-DGGE coincided with those found among the cultured isolates. CONCLUSIONS: Ammonia-assimilating micro-organisms obtained by the cultivation method were not predominant in the microbial community during the composting process: however certain cultured actinomycetes were members of predominant species in the actinomycetes community. SIGNIFICANCE AND IMPACT OF THE STUDY: Ammonia assimilation by micro-organisms is one of the important mechanisms for ammonia retention in the composting process. Cultivable actinomycetes are a means for preventing ammonia emission from the composting process.     

A swine toll-like receptor 2-expressing transfectant as a potential primary screening system for immunobiotic microorganisms

Toll-like receptor 2 (TLR2) has been shown to mediate cell signaling in response to microbial cell wall components, such as peptidoglycan, lipoteichoic acid, microbial lipoprotein, and zymosan. In this study, we cloned the swine TLR2 and used it to transfect Chinese hamster ovary K-1 cells. We demonstrated that the swine TLR2-expressing transfectant can bind not only zymosan from yeast cell wall components but also intact lactic acid bacteria, resulting in the activation of nuclear factor-kappaB. These findings suggest that the swine TLR2-expressing transfectant can be very useful for the primary screening of immunobiotic microorganisms.     

Augmentation of T(H)-1 type response by immunoactive AT oligonucleotide from lactic acid bacteria via Toll-like receptor 9 signaling

Toll-like receptor 9, which is expressed on the surface of antigen presenting cells and which was recently identified in the cytoplasmic follicle, recognizes bacterial CpG oligodeoxynucleotides (ODNs), resulting in the induction of a potent immune response. However, in our previous study, we found that TLR9 potentially recognizes not only CpG ODN but also non-CpG ODN such as AT ODN. Therefore, in the present study, to investigate this possibility, we elucidated the effects of AT ODN on T(H)-1, T(H)-2 type cytokine induction via TLR9 by real-time quantitative PCR analysis and ELISA of the swine TLR9 transfectant. The results demonstrated that the T(H)-1 type cytokines such as interleukin (IL)-12p70 and interferon (IFN)-gamma were strongly induced by AT ODN compared to the unexposed controls, while T(H)-2 type cytokines were not induced. These results indicate that the AT ODN can augment the T(H)-1 immune response, which plays an important role in prevention of allergic responses. Moreover, the swine TLR9 transfectant demonstrated its usefulness for evaluation of immunostimulation by bacterial DNA through the detection of T(H)-1, T(H)-2 type cytokine induction via TLR9 signaling.     

肠道共生细菌预发酵鸡粪对黑水虻生长发育的影响

以利用黑水虻Hermetia illucens L.肠道共生细菌不同菌株培养液预发酵的鸡粪为培养料,研究了添加不同菌株的发酵鸡粪对黑水虻幼虫生长发育的影响。结果表明,添加不同菌株的预发酵鸡粪能显著增加预蛹和蛹的重量,显著提高化蛹率并增加雌、雄成虫体长,缩短预蛹所需时间,但对幼虫存活率、成虫羽化率、成虫寿命没有显著影响。同时,不同菌株的作用效果有所不同。     

Root Growth Effects on Soluble C and P in Manured and Non-manured Soils

There is limited research on relationships between root characteristics and soil chemical properties and processes. Because previous studies have shown specific C compounds may release previously sorbed P and make P more plant-available, crops which contribute to high soil C levels could play an important role in soil P cycling. The objectives of this study were to determine (1) whether rotation crops had different amounts of root growth, (2) whether different amounts of root growth among the crop species could be related to different levels of soluble soil C and (3) whether there were differences in P concentration among the soils under different crops that could be related to soluble C soil concentration. Roots and soil from potato (Solanum tuberosum L.), barley (Hordeum vulgare L.), soybean (Glycine max (L.) Merr.), and a forage consisting of alfalfa (Medicago sativa L.) and timothy (Phleum pretense L.) were sampled from the Aroostook Research Farm in Presque Isle, Maine, during the summers of 2003 and 2004 to determine root length density (RLD) and soluble C and P concentrations. Half of the sampled plots were amended with beef manure and half were not amended. Barley and forage consistently had higher RLD than potato or soybean crops. Barley and forage typically had higher concentrations of soluble soil C than potato or soybean, but the differences were significant at only three of the five sampling dates. RLD was significantly correlated to soluble C (r=0.56) only for amended soils on the August 2003 sampling date. For other dates r values were non-significant and ranged from 0.32 to 0.49. As with soil C, soluble soil P levels were typically higher in barley and forage than in potato or soybean crops. Significant differences were detected at four of the five sampling dates. Correlations between soluble C and soluble P were significant at two of the five sampling dates (r = 0.58 and 0.62) in amended soils and one of five sampling dates (r = 0.80) in unamended soils. Although the correlations between RLD and soluble C were not significant at every sampling date, the August 2003 data do suggest a possible effect of roots on soluble C. In addition, significant correlations between soluble C and soluble P at several sampling dates suggest a relationship between these parameters. Therefore cropping systems that include crops with higher amounts of root growth may promote increased soluble soil C levels and enhance P bioavailability.     

Characterization of a distinctive pattern of periovulatory leptin secretion and its relationship with ovulation rate and luteal function in swine with obesity/leptin resistance

Patterns of leptin secretion during the estrous cycle and the possible relationship of changes in circulating leptin during the periovulatory period with ovarian function in sows of obese (Iberian breed) and lean genotype (Large White×Landrace) were evaluated in two consecutive experiments. Plasma leptin concentrations throughout the estrous cycle in lean sows remain unchanged, but Iberian females showed a periovulatory increase in circulating leptin levels without associated changes in body condition and fatness. In these sows, plasma leptin concentrations at Days -1 and 0 of the cycle were found to be positively correlated with the ovulation rate (r=0.943 and r=0.987, respectively; P<0.05 for both), but the levels of leptin at Day 0 were negatively correlated with the progesterone release from Day 3 (r=-0.557; P<0.05) and, became more evident at Day 5 of the estrous cycle (r=-0.924; P<0.005). Such relationships were not observed in the females of the lean genotype. In conclusion, the present study indicates the existence of a distinctive pattern in the periovulatory leptin secretion in swine with obesity and leptin resistance, which is associated with the number and functionality of the corpora lutea present in the subsequent cycle.     

‘Bioenergy from cattle manure? Implications of anaerobic digestion and subsequent pyrolysis for carbon and nitrogen dynamics in soil’

Cattle manure can be processed to produce bioenergy, resulting in by‐products with different physicochemical characteristics. To evaluate whether application of such bioenergy by‐products to soils would be beneficial compared with their unprocessed counterpart, we quantified differences in greenhouse gas emissions and carbon (C) and nitrogen (N) dynamics in soil. Three by‐products (¹⁵N‐labeled cattle manure, from which anaerobic digestate was obtained, which was subsequently pyrolysed) were applied to a loess and a sandy soil in a laboratory incubation study. The highest losses of soil C from biological activity (CO₂ respiration) were observed in manure treatments (39% and 32% for loess and sandy soil), followed by digestate (31% and and 18%), and biochar (15% and and 7%). Emissions of nitrous oxide (N₂O) ranged from 0.6% of applied N from biochar to 4.0% from manure. Isotope labeling indicated that manure N was most readily mineralized, contributing 50% to soil inorganic N. The anaerobic digestate was the only by‐product increasing the mineral N pool, while reducing emissions of N₂O compared with manure. In biochar treatments, less than 18.3% of soil mineral N derived from the biochar, while it did not constrain mineralization of native soil N. By‐products of anaerobic digestion and pyrolysis revealed soil fertility in addition to environmental benefits. However, the reported advantages lessen when the declining yields of C and N over the bioenergy chain are considered.