Stratification of solids,nitrogen and phosphorus in swine manure in deep pits under slatted floors

Manure slurries stored in pits under slatted floors of both finishing and nursery barns were sampled at four different depths to study stratifications of total solids (TS) and nutrients (nitrogen and phosphorus), and to determine the relationship between the stratification of TS and nutrients. The results obtained can be used to improve the management and handling of swine manure in the under-slat storage pits. A management scheme that can be adopted for both the finishing and the nursery barns' pits is the layer-by-layer harvesting of the manure. The thinner manure, which is lower in nutrients, can be spread on land near the production units in larger volumes or it may pumped to land remote from the production units without causing many clogging problems. The thicker manure, higher in nutrients, can be transported to land further away and spread in smaller volumes. The TS content of each stratum can be used to accurately estimate the nitrogen and phosphorus levels in the respective strata so that application rates can easily be adjusted accordingly during the time of land application.     

Using Plasmodium knowlesi as a model for screening Plasmodium vivax blood-stage malaria vaccine targets reveals new candidates

Plasmodium vivax is responsible for the majority of malaria cases outside Africa. Unlike P. falciparum, the P. vivax life-cycle includes a dormant liver stage, the hypnozoite, which can cause infection in the absence of mosquito transmission. An effective vaccine against P. vivax blood stages would limit symptoms and pathology from such recurrent infections, and therefore could play a critical role in the control of this species. Vaccine development in P. vivax, however, lags considerably behind P. falciparum, which has many identified targets with several having transitioned to Phase II testing. By contrast only one P. vivax blood-stage vaccine candidate based on the Duffy Binding Protein (PvDBP), has reached Phase Ia, in large part because the lack of a continuous in vitro culture system for P. vivax limits systematic screening of new candidates. We used the close phylogenetic relationship between P. vivax and P. knowlesi, for which an in vitro culture system in human erythrocytes exists, to test the scalability of systematic reverse vaccinology to identify and prioritise P. vivax blood-stage targets. A panel of P. vivax proteins predicted to function in erythrocyte invasion were expressed as full-length recombinant ectodomains in a mammalian expression system. Eight of these antigens were used to generate polyclonal antibodies, which were screened for their ability to recognize orthologous proteins in P. knowlesi. These antibodies were then tested for inhibition of growth and invasion of both wild type P. knowlesi and chimeric P. knowlesi lines modified using CRISPR/Cas9 to exchange P. knowlesi genes with their P. vivax orthologues. Candidates that induced antibodies that inhibited invasion to a similar level as PvDBP were identified, confirming the utility of P. knowlesi as a model for P. vivax vaccine development and prioritizing antigens for further follow up.     

Modeling effects of granules on the start-up of anaerobic digestion of dairy wastewater with Langmuir and extended Freundlich equations

The effects of granules-inocula on the start-up of anaerobic reactors treating dairy manure were studied in a batch-fed reactor. The effects of start-up period and ratio of granules to feed were analyzed. Results indicated that the effects of start-up period could be described by Langmuir model, while the Extended Freundlich model could be used to model the effects of ratio of granules to feed on cumulative biogas production. In addition, transmission electron microscopes (TEM) and scanning electron microscope analysis were conducted to elucidate the distribution of microbial population and micro-colonies in granules and manure. From the TEM micrographs analyses, the ratios the Syntrophobacter and methanogens in granule and manure were shown to be 1.57 ± 0.42 and 0.22 ± 0.20, respectively. These results demonstrated that granules-inocula could reduce the period required for onset of biogas by 25%.     

Caenorhabditis elegans development requires mitochondrial function in the nervous system

The mitochondrial respiratory chain (MRC) supplies the majority of the energy requirements of most eucaryotic cells. A null mutation in the Caenorhabditis elegans nuo-1 gene encoding a subunit of complex I (NADH-ubiquinone oxidoreductase) is lethal, leading to a developmental arrest at the third larval stage. To identify the tissues that regulate development in response to mitochondrial dysfunction, we restored nuo-1 expression with tissue-specific promoters. Only expression of nuo-1 ubiquitously or in the nervous system supported development to the adult stage. Pharyngeal expression of nuo-1 allowed development to proceed to the fourth larval stage. Expression of nuo-1 in the body muscles or in the germline had no effect. Furthermore, only ubiquitous or nervous system expression of nuo-1 allowed exit from the dauer state. Our results indicate that MRC function in the nervous system is needed to send and receive signals that control larval development and exit from dauer.     

Dynamics of the Developing Chick Chorioallantoic Membrane Assessed by Stereology,Allometry, Immunohistochemistry and Molecular Analysis

The chick chorioallantoic membrane (CAM) is a widely used model for the study of angiogenesis, tumour growth, as well as drug efficacy. In spite of this, little is known about the developmental alteration from its appearance to the time of hatching. In the current study the CAM has been studied by classical stereology and allometry. Expression levels of selected angiogenesis-related molecules were estimated by RT-PCR and cell dynamics assessed by proliferation and apoptosis assays. Absolute CAM volume increased from a low of 0.47 ± 0.11 cm³ at embryonic day 8 (E8) to a high of 2.05 ± 0.27 cm³ at E18, and then decreased to 1.6 ± 0.47 cm³ at E20. On allometric analysis, three growth phases were identifiable. Between E8-13 (phase I), the CAM grew fastest; moderately in phase II (E13-18) but was regressing in phase III (E18-20). The chorion, the mesenchyme and the allantoic layers grew fastest in phase I, but moderately in phase II. The mesenchyme grew slowly in phase III while the chorion and allantois were regressing. Chorionic cell volume increased fastest in phase I and was regressing in phase III. Chorionic capillaries grew steadily in phase I and II but regressed in phase III. Both the chorion and the allantois grew by intrinsic cell proliferation as well as recruitment of cells from the mesenchyme. Cell proliferation was prominent in the allantois and chorion early during development, declined after E17 and apoptosis started mainly in the chorion from E14. VEGFR2 expression peaked at E11 and declined steadily towards E20, VEGF peaked at E13 and E20 while HIF 1α had a peak at E11 and E20. Studies targeting CAM growth and angiogenesis need to take these growth phases into consideration     

Decanal as a major component of larval aggregation pheromone of the greater wax moth,Galleria mellonella

Larvae of the greater wax moth (GWM), Galleria mellonella, a destructive pest of the honeybee (Apis mellifera), have been observed to display aggregation behaviours. However, the underlying mechanism by which these larvae come together remains unknown. We hypothesized that the GWM larvae detect, orient towards and utilize conspecific larval chemical cues to aggregate in groups. We used dual‐choice olfactometer assays to investigate the involvement of conspecific larval odours in their aggregation amongst 3–5th instar and 8th instar larvae. The assays revealed that only 8th instar larvae were significantly attracted to their odours and those emanating from newly spun cocoons. Coupled gas chromatography–mass spectrometry (GC‐MS) of larval head space odours analysis revealed the presence of four compounds: nonanal, decanal, tridecane and tetradecane in pupal and mature larval odour extracts. However, using synthetic compounds, behavioural assays showed that only decanal induced significant attraction, therefore, suggesting its role as a major component of the larval aggregation pheromone of GWM. Our findings reveal the involvement of volatile organic compounds in the aggregation behaviour of mature wax moth larvae and thereby offer prospects for the development of an odour‐baited in‐hive trapping management tool for wax moth larva.     

Influence of C18 long chain fatty acids on hydrogen metabolism

During anaerobic treatment, several microorganisms mediate a series of reactions to convert reduced compounds (electron donors) into methane. Inhibitors such as long chain fatty acids (LCFAs) can affect several anaerobic microbial populations and decrease the treatment efficiency. The effects of three C18 LCFAs on hydrogenotrophic methanogens in a flocculated mixed anaerobic culture were assessed in this study. The reaction half-life and the hydrogen versus time profiles were used to characterize the inhibition process. The half-life values and profiles were similar for controls and cultures exposed to LCFAs for 1 h. The hydrogen inhibition was a function of the exposure time and the LCFA concentration except for cultures exposed to stearic acid (SA). A statistical analysis of the reaction half-life for cultures incubated with 1,500 and 2,000 mg L(-1) LCFAs for 48 h, revealed the following inhibition trend: linoleic acid (LA) > oleic acid (OA) > SA. After 48 h of exposure, no clear inhibition trend was observed for cultures inoculated with LCFA mixtures; however, at levels of 1,500 and 2,000 mg L(-1), the reaction half-life values were less than that observed for cultures fed with only LA. Based on the reaction half-life data, all of the LCFAs except SA at threshold levels of approximately 1,500 mg L(-1) inhibited hydrogen metabolism. The greatest inhibition and, hence, the largest amount of accumulated hydrogen was observed in cultures fed with 2,000 mg L(-1) LA and incubated for 48 h.     

Structural analysis of enzyme-resistant isomaltooligosaccharides reveals the elongation of α-(1→3)-linked branches in Weissella confusa dextran

Weissella confusa VTT E-90392 is an efficient producer of a dextran that is mainly composed of α-(1→6)-linked D-glucosyl units and very few α-(1→3) branch linkages. A mixture of the Chaetomium erraticum endodextranase and the Aspergillus niger α-glucosidase was used to hydrolyze W. confusa dextran to glucose and a set of enzyme-resistant isomaltooligosaccharides. Two of the oligosaccharides (tetra- and hexasaccharide) were isolated in pure form and their structures elucidated. The tetrasaccharide had a nonreducing end terminal α-(1→3)-linked glucosyl unit (α-D-Glcp-(1→3)-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc), whereas the hexasaccharide had an α-(1→3)-linked isomaltosyl side group (α-D-Glcp-(1→6)[α-D-Glcp-(1→6)-α-D-Glcp-(1→3)]-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc). A mixture of two isomeric oligosaccharides was also obtained in the pentasaccharide fraction, which were identified as (α-D-Glcp-(1→6)-α-D-Glcp-(1→3)-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc) and (α-D-Glcp-(1→6)[α-D-Glcp-(1→3)]-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc). The structures of the oligosaccharides indicated that W. confusa dextran contains both terminal and elongated α-(1→3)-branches. This is the first report evidencing the presence of elongated branches in W. confusa dextran. The (1)H and (13)C NMR spectroscopic data on the enzyme-resistant isomaltooligosaccharides with α-(1→3)-linked glucosyl and isomaltosyl groups are published here for the first time.     

Manure sampling procedures and nutrient estimation by the hydrometer method for gestation pigs

Three manure agitation procedures were examined in this study (vertical mixing, horizontal mixing, and no mixing) to determine the efficacy of producing a representative manure sample. The total solids content for manure from gestation pigs was found to be well correlated with the total nitrogen (TN) and total phosphorus (TP) concentrations in the manure, with highly significant correlation coefficients of 0.988 and 0.994, respectively. Linear correlations were observed between the TN and TP contents and the manure specific gravity (correlation coefficients: 0.991 and 0.987, respectively). Therefore, it may be inferred that the nutrients in pig manure can be estimated with reasonable accuracy by measuring the liquid manure specific gravity. A rapid testing method for manure nutrient contents (TN and TP) using a soil hydrometer was also evaluated. The results showed that the estimating error increased from +/-10% to +/-30% with the decrease in TN (from 1000 to 100 ppm) and TP (from 700 to 50 ppm) concentrations in the manure. Data also showed that the hydrometer readings had to be taken within 10 s after mixing to avoid reading drift in specific gravity due to the settling of manure solids.