Serotonin-induced hypophagia is mediated via α2 and ß2 adrenergic receptors in neonatal layer-type chickens.

1. Serotoninergic and adrenergic systems play crucial roles in feed intake regulation in avians but there is no report on possible interactions among them. So, in this study, 5 experiments were designed to evaluate the interaction of central serotonergic and adrenergic systems on food intake regulation in 3 h food deprived (FD3) neonatal layer-type chickens. 2. In Experiment 1, chickens received intracerebroventricular (ICV) injection of control solution, serotonin (56.74 nmol), prazosin (α1 receptor antagonist, 10 nmol) and co-injection of serotonin plus prazosin. In Experiment 2, control solution, serotonin (56.74 nmol), yohimbine (α2 receptor antagonist, 13 nmol) and co-injection of serotonin plus yohimbine were used. In Experiment 3, the birds received control solution, serotonin (56.74 nmol), metoprolol (ß1 receptor antagonist, 24 nmol) and co-injection of serotonin plus metoprolol. In Experiment 4, injections were control solution, serotonin (56.74 nmol), ICI 118.551 (ß2 receptor antagonist, 5 nmol) and serotonin plus ICI 118.551. In Experiment 5, control solution, serotonin (56.74 nmol), SR59230R (ß3 receptor antagonist, 20 nmol) and co-administration of serotonin and SR59230R were injected. In all experiments the cumulative food intake was measured until 120 min post injection. 3. The results showed that ICV injection of serotonin alone decreased food intake in chickens. A combined injection of serotonin plus ICI 118.551 significantly attenuated serotonin-induced hypophagia. Also, co-administration of serotonin and yohimbine significantly amplified the hypophagic effect of serotonin. However, prazosin, metoprolol and SR59230R had no effect on serotonin-induced hypophagia in chickens. 4. These results suggest that serotonin-induced feeding behaviour is probably mediated via α2 and ß2 adrenergic receptors in neonatal layer-type chicken.

Cultivation and qPCR Detection of Pathogenic and Antibiotic-Resistant Bacterial Establishment in Naive Broiler Houses.

Conventional commercial broiler production involves the rearing of more than 20,000 broilers in a single confined space for approximately 6.5 wk. This environment is known for harboring pathogens and antibiotic-resistant bacteria, but studies have focused on previously established houses with mature litter microbial populations. In the current study, a set of three naive houses were followed from inception through 11 broiler flocks and monitored for ambient climatic conditions, bacterial pathogens, and antibiotic resistance. Within the first 3 wk of the first flock cycle, 100% of litter samples were positive for and , whereas was cultivation negative but PCR positive. Antibiotic resistance genes were ubiquitously distributed throughout the litter within the first flock, approaching 10 to 10 genomic units g. Preflock litter levels were approximately 10 CFU g for heterotrophic plate count bacteria, whereas midflock levels were >10 colony forming units (CFU) g; other indicators demonstrated similar increases. The influence of intrahouse sample location was minor. In all likelihood, given that preflock levels were negative for pathogens and antibiotic resistance genes and 4 to 5 Log lower than flock levels for indicators, incoming birds most likely provided the colonizing microbiome, although other sources were not ruled out. Most bacterial groups experienced a cyclical pattern of litter contamination seen in other studies, whereas microbial stabilization required approximately four flocks. This study represents a first-of-its-kind view into the time required for bacterial pathogens and antibiotic resistance to colonize and establish in naive broiler houses.

A new sampler for stratified lagoon chemical and microbiological assessments.

A sampler was needed for a spatial and temporal study of microbial and chemical stratification in a large swine manure lagoon that was known to contain zoonotic bacteria. Conventional samplers were limited to collections of surface water samples near the bank or required a manned boat. A new sampler was developed to allow simultaneous collection of multiple samples at different depths, up to 2.3 m, without a manned boat. The sampler was tethered for stability, used remote control (RC) for sample collection, and accommodated rapid replacement of sterile tubing modules and sample containers. The sampler comprised a PVC pontoon with acrylic deck and watertight enclosures, for a 12 VDC gearmotor, to operate the collection module, and vacuum system, to draw samples into reusable autoclavable tubing and 250-mL bottles. Although designed primarily for water samples, the sampler was easily modified to collect sludge. The sampler held a stable position during deployment, created minimal disturbance in the water column, and was readily cleaned and sanitized for transport. The sampler was field tested initially in a shallow fresh water lake and subsequently in a swine manure treatment lagoon. Analyses of water samples from the lagoon tests showed that chemical and bacterial levels, pH, and EC did not differ between 0.04, 0.47, and 1.0 m depths, but some chemical and bacterial levels differed between winter and spring collections. These results demonstrated the utility of the sampler and suggested that future manure lagoon studies employ fewer or different depths and more sampling dates.

Runoff quality from no-till cotton fertilized with broiler litter in subsurface bands.

Surface broadcast of broiler litter to no-till row crops exposes the litter and its nutrients to risks of loss in runoff water and volatilization and may limit the potential benefit of litter to the crops. Subsurface banding of litter could alleviate these risks. A field study was conducted in 2008 and 2009 on an upland Falkner silt loam soil to determine the effect of broiler litter placement on runoff nutrient losses from no-till cotton ( L.). Treatments included surface broadcast broiler litter applied manually, subsurface-banded litter applied by tractor-drawn equipment, and no broiler litter, all in combination with or without winter wheat ( L.) cover crop residue. Broiler litter rate was 5.6 Mg ha. The experimental design was a randomized complete block with a split-plot arrangement of treatments replicated three times. In 2008, simulated rainfall was used to generate runoff 27 d after litter application. Subsurface-banded litter reduced runoff total C, N, P, NH, NO, Cu, Zn and water-soluble P (WP) concentrations by 72, 64, 51, 49, 70, 36, 65, and 77%, respectively, compared with surface broadcast. The reductions were greater in 2009 where runoff occurred 1 d after litter application. Bacterial runoff was decreased by one log with subsurface-banded litter compared to surface broadcast. Except for C, NH, N, and WP, the presence of winter cover crop residue did not affect the load or runoff nutrient concentrations in either year. The results indicate that subsurface banding litter to no-till cotton substantially reduces nutrient and bacterial losses in runoff compared with surface broadcasting.

The effect of poultry manure application rate and AlCl(3) treatment on bacterial fecal indicators in runoff.

Increasing costs associated with inorganic fertilizer have led to widespread use of broiler litter. Proper land application, typically limiting nutrient loss, is essential to protect surface water. This study was designed to evaluate litter-borne microbial runoff (heterotrophic plate count bacteria, staphylococci, Escherichia coli, enterococci, and Clostridium perfringens) while applying typical nutrient-control methods. Field studies were conducted in which plots with high and low litter rates, inorganic fertilizer, AlCl(3)-treated litter, and controls were rained on five times using a rain generator. Overall, microbial runoff from poultry litter applied plots was consistently greater (2-5 log(10) plot(-1)) than controls. No appreciable effect on microbial runoff was noted from variable litter application rate or AlCl(3) treatments, though rain event, not time, significantly affected runoff load. C. perfringens and staphylococci runoff were consistently associated with poultry litter application, during early rain events, while other indicators were unreliable. Large microbial runoff pulses were observed, ranging from 10(2) to 10(10) CFU plot(-1); however, only a small fraction of litter-borne microbes were recoverable in runoff. This study indicated that microbial runoff from litter-applied plots can be substantial, and that methods intended to reduce nutrient losses do not necessarily reduce microbial runoff.

Single-step real-time PCR to quantify hepatitis B virus and distinguish genotype D from non-D genotypes.

Hepatitis B virus (HBV) viral load and its genotype play important roles in clinical outcome, management of disease and response to antiviral therapy. In many parts of the world such as Europe or the Middle East, distinguishing HBV genotype D from non-D is most relevant for treatment decisions, because genotype D-infected patients respond poorly to interferon-based therapeutic regimens. Here, we developed an in-house real-time PCR to concordantly assess HBV genotype (D vs non-D) based on melt curve analysis and quantify the viral load. Genotype distinction was established with control plasmids of all HBV genotypes and validated with 57 clinical samples from patients infected with six different HBV genotypes. Our in-house real-time PCR assay could discriminate HBV genotype D from non-D using single-step melt curve analysis with a 2 °C difference in the melt curve temperature in all samples tested. Viral load quantification was calibrated with the WHO HBV international standard, demonstrating an excellent correlation with a commercial kit (r = 0.852; P < 0.0001) in a linear range from 3.2 × 10(2) to 3.2 × 10(10) IU/mL. In conclusion, we developed a rapid, simple and cost-effective method to simultaneously quantify and distinguish HBV genotypes D from non-D with a single-step PCR run and melt curve analysis. This assay should be a useful diagnostic alternative to aid clinical decisions about initiation and choice of antiviral therapy, especially in geographical regions with a high prevalence of HBV genotype D.

Effect of surface incorporation of broiler litter applied to no-till cotton on runoff quality.

Surface application of broiler litter to no-till cotton could lead to degradation of water quality. Incorporation of broiler litter into the top surface soil (0.05 m) could alleviate this risk. A 2-yr field study was conducted on a silt loam upland soil to determine the effect of incorporation of broiler litter into the soil surface on nutrient and bacterial transport in runoff. The experimental design was a randomized complete block with four treatments and three replications. Treatments were (i) unfertilized control; (ii) surface-appliedbroiler litter at 7.8 Mg ha(-1) without incorporation; (iii) surface-applied broiler litter at 7.8 Mg ha(-1) with immediate incorporation; and (iv) inorganic fertilizer N (urea ammonium nitrate, 32% N) and inorganic fertilizer P (triple superphosphate) at the recommended rate. Phosphorus was surface appliedat 25 kg ha(-1) and N was injected at 101 kg ha(-1) into the soil using a commercial liquid fertilizer applicator. Runoff was collected from small runoff plots (2.4 m by 1.6 m) established at the bottom side of main plots (13.7 m by 6.0 m). Incorporation of broiler litter reduced total N (TN), NO3-N, water soluble P (WSP), and total P (TP) concentrations in runoffby 35, 25, 61, and 64%, respectively, and litter-associated bacteria by two to three orders of magnitude compared with unincorporated treatment. No significant difference in total suspended solids (TSS) in runoffwas obtained between incorporated and unincorporated treatments. Incorporation of broiler litter into the surface soil in the no-till system immediately after application minimized the potential risk for surface nutrient losses and bacteria transport in runoff.

Laboratory and field evaluation of broiler litter nitrogen mineralization.

Two studies were conducted for this research. First, a laboratory incubation to quantify broiler litter N mineralization with the following treatments: two soil moisture regimes, constant at 60% water fill pore space (WFPS) and fluctuating (60-30% WFPS), three soil types, Brooksville silty clay loam, Ruston sandy loam from Mississippi, and Catlin silt loam from Illinois. Second, a field incubation study to quantify broiler litter N mineralization using similar soils and litter application rates as the laboratory incubation. Broiler litter was applied at an equivalent rate of 350 kg total N ha(-1) for both studies except for control treatments. Subsamples were taken at different timing for both experiments for NO3-N and NH4-N determinations. In the laboratory experiment, soil moisture regimes had no significant impact on litter-derived inorganic N. Total litter-derived inorganic N across all treatments increased from 23 mg kg(-1) at time 0, to 159 mg kg(-1) at 93 d after litter application. Significant differences were observed among the soil types. Net litter-derived inorganic N was greater for Brooksville followed by Ruston and Catlin soils. For both studies and all soils, NH4-N content decreased while NO3-N content increased indicating a rapid nitrification of the mineralized litter N. Litter mineralization in the field study followed the same trend as the laboratory study but resulted in much lower net inorganic N, presumably due to environmental conditions such as precipitation and temperature, which may have resulted in more denitrification and immobilization of mineralized litter N. Litter-derived inorganic N from the field study was greater for Ruston than Brooksville. Due to no impact by soil moisture regimes, additional studies are warranted in order to develop predictive relationships to quantify broiler litter N availability.

Swine effluent application timing and rate affect nitrogen use efficiency in common bermudagrass.

Bermudagrass [Cynodon dactylon (L.) Pers.] hay production is integral to manure management on southeastern swine farms. But swine effluent timing must be synchronized with crop nitrogen (N) demands to decrease the potential for soil N accumulation and nitrate (NO(3)) leaching. Field studies were conducted on a Prentiss sandy loam (coarse-loamy, siliceous, semiactive, thermic Glossic Fragiudult) to determine N-use efficiency (NUE) and residual soil NO(3)-N. Two rates of 10 and 20 cm yr(- 1) ( approximately 260 and 480 kg ha(-1) N, respectively) were applied in four timing treatments: April to September (full season), April to May, June to July, and August to September. Plots were harvested every 7 to 9 wk beginning in June, and soil was sampled in fall after a killing frost and the following spring. Annual uptake of N and P were least in the August to September timing treatment. Doubling the effluent rate increased N uptake 112% in 2000 (from 130 to 276 kg ha(-1)) and 53% in 2001 (from 190 to 290 kg ha(-1)), suggesting 10-cm did not meet crop N demands. Due to low rainfall and decreased forage yield in 2000, doubling the effluent rate led to increased soil NO(3)-N to 30-cm depth in fall 2000 and spring 2001. Averaged across timing treatments, soil NO(3)-N at 5-cm depth ranged from 8.5 mg kg(-1) in non-irrigated controls to 39.6 mg kg(-1) with 20-cm effluent. Results indicate low NUE in the order of 30 to 38% for applications in August to September increase the risk to surface and ground water quality from excess N remaining in soil.

Using broiler litter and swine manure lagoon effluent in sawdust-based swine mortality composts: Effects on nutrients, bacteria, and gaseous emissions.

Disposition of mortalities challenges confined animal feeding operations (CAFOs), especially sow (farrowing) farms, which experience mortalities daily. Regulations and transportation costs may preclude incineration, landfill burial, and rendering; therefore, swine CAFOs in Mississippi in the Mid-South U.S. often compost mortalities. In this study, a farm-standard composting mix of sawdust (S) and water (W) was compared with mixes where N was supplied by broiler litter (L) and water was replaced with swine lagoon effluent (E). The objective was to assess the effects of these manure byproducts: 1) on nutrients and bacteria in composts destined for land application; and 2) on emissions of ammonia and greenhouse gases. Three replications of four mixes (SW, SLW, SE, SLE) were compared in microcosms comprising modified plastic recycling bins. The experiment was repeated three times in different seasons in one year. Mixes were compared for differences in temperature, water content, nutrients (C, N, P, K, Ca, Mg, Na, Mn, Fe, Cu, Zn), bacteria (Gram-, Gram+, Clostridium perfringens, Salmonella, Listeria, Escherichia coli), and emissions (NH3, CO2, CH4, N2O). Litter addition increased composting temperatures initially and after aerations; increased nutrient concentrations, except C, in start mixes and all except C and N, in finish mixes; increased Gram+ bacteria, Salmonella, and E. coli in start mixes, but only Gram+s in finish mixes; and increased emissions. Effluent addition increased early composting temperatures; had no effect on nutrients or bacteria, except increased C. perfringens in start, but not finish mixes; and had no effect on emissions. Nutrients in finish composts did not differ among mixes for N (average 3.3%), but litter composts had more P and K, and lower N:P than composts without litter. Improving mortality composting is of global importance as increasing livestock populations and intensive animal production systems require practical, safe, environmentally sound disposal of carcasses.

Swine effluent irrigation rate and timing effects on bermudagrass growth, nitrogen and phosphorus utilization, and residual soil nitrogen.

Maximizing utilization of effluent nutrients by forage grasses requires a better understanding of irrigation rate and timing effects. This study was conducted in 1998 and 1999 on a Vaiden silty clay (very-fine, smectitic, thermic Aquic Dystrudert) soil to determine the effects of swine lagoon effluent irrigation rate and timing on bermudagrass [Cynodon dactylon (L.) Pers.] growth, nitrogen (N) and phosphorus (P) recovery, and postseason soil profile NO3(-)-N. Treatments consisted of swine effluent irrigation at the rates of 0, 5, 10, 15, and 20 ha-cm. Two additional treatments included 2.5 ha-cm applied on 1 September and 1 October in addition to a base summer rate of 10 ha-cm. In both years for early to mid-season irrigation, bermudagrass dry matter yield quadratically increased with increasing swine effluent irrigation rates. Averaged across years, effluent irrigation in October resulted in 30% less dry matter than in September. For late-season irrigation, apparent N recovery averaged 59% less and P recovery averaged 46% less with a delay in irrigation from 1 September to 1 October. The greatest quantity of soil NO3(-)-N was associated with both the greatest effluent rate and October irrigation treatments. Minimal yield benefit was obtained when effluent was applied at rates greater than 10 ha-cm during the summer months. Late-season irrigation, especially after 1 October for areas with similar climatic conditions, should be avoided to maximize synchronization of nutrient availability with maximum growth rates to minimize potential offsite movement of residual soil N and P.