Identification of biomarkers associated with ferroptosis in diabetic retinopathy based on WGCNA and machine learning.

Objective: Diabetic retinopathy (DR) is a chronic progressive eye disease that affects millions of diabetic patients worldwide, and ferroptosis may contribute to the underlying mechanisms of DR. The main objective of this work is to explore key genes associated with ferroptosis in DR and to determine their feasibility as diagnostic markers. Methods: WGCNA identify the most relevant signature modules in DR. Machine learning methods were used to de-screen the feature genes. ssGSEA calculated the scoring of immune cells in the DR versus control samples and compared the associations with the core genes by Spearman correlation. Results: We identified 2,897 differential genes in DR versus normal samples. WGCNA found tan module to have the highest correlation with DR patients. Finally, 20 intersecting genes were obtained from differential genes, tan module and iron death genes, which were screened by LASSO and SVM-RFE method, and together identified 6 genes as potential diagnostic markers. qPCR verified the expression and ROC curves confirmed the diagnostic accuracy of the 6 genes. In addition, our ssGSEA scoring identified these 6 core genes as closely associated with immune infiltrating cells. Conclusion: In conclusion, we analyzed for the first time the potential link of iron death in the pathogenesis of DR. This has important implications for future studies of iron death-mediated pro-inflammatory immune mechanisms.

Research Note: Evaluation of the efficacy of engineered water nanostructures in inactivating airborne bacteria in poultry houses.

Methods to control microbial contamination in confined livestock facilities are important to the health of both animals and workers. In addition, bacterial contamination is also a food safety issue. The most common disinfection technique employed in livestock facilities is the application of oxidizing agents (e.g., potassium peroxymonosulphate, chlorine, hydrogen peroxide, ozone). However, these techniques are associated with a number of limitations (e.g., toxicity, high cost, corrosiveness). Recently, engineered water nanostructures (EWNS) generated using an electrospray system was found effective in inactivating foodborne bacteria. Thus, this study investigated the efficacy of EWNS generated using a laboratory-scale electrospray system in inactivating bacteria found in poultry facilities. The effects of various operating conditions (distance between the injector and grounded electrode of the electrospray system, applied voltage, liquid pH and conductivity, liquid flow rate, and treatment time) on the efficacy were also assessed. In these various experiments, airborne bacterial samples were collected from a pullet room using tryptic soy agar plates and then exposed to EWNS under varying conditions. After treatment, the plates were incubated at 37°C prior to colony counting. Reductions in bacterial concentrations up to 1.26 logs were obtained. The results indicate that the EWNS generated by the electrospray system can be a potential chemical-free alternative to conventional disinfection methods. Future tests will focus on scaling up the system for larger scale trials.

Diurnal and seasonal variations of odor emissions from broiler and cage-layer barns in the Canadian Prairies.

Odor concentrations (OC) and emissions (OE) were measured for a commercial broiler barn and a cage-layer barn in a cold region (the Canadian Prairies). Seasonal OC and OE profiles were plotted by monthly measurements over the course of a year from March 2015 to February 2016, and diurnal profiles were generated by 2-day measurements in cold, mild, and warm seasons, respectively. Seasonal OC and OE varied for both barns; OC was higher in the cold season, but OE was higher in the mild and warm seasons. The broiler barn had higher annual average OC (718 OU m-3) but slightly lower annual average OE (127 OU s-1 AU-1; AU is per 500 kg of body mass) than the layer barn (574 OU m-3 and 140 OU s-1 AU-1). For the layer barn, OC and OE were reduced by 31% and 33% in the cold season and by 30% and 26% in the mild season after manure removal compared with before manure removal. Statistical results showed increased outdoor temperature and ventilation rate (VR) were associated with decreased OC but increased OE for both barns. Finally, both single linear and multi-linear regression models of OE were developed.

Toxic gas and respirable dust concentrations and emissions from broiler and cage-layer barns in the Canadian Prairies.

This study monitored concentrations and emissions of ammonia (NH3), hydrogen sulfide (H2S), and respirable dust for a commercial broiler and a cage-layer barn in the Canadian Prairies over a year between March 2015 and February 2016. Seasonal concentration and emission profiles were acquired by monthly measurements, while diurnal profiles were generated in different seasons. The indoor air quality was evaluated considering both the individual and the additive health effect (respiratory irritation) of the three air pollutants. In winter, both 8-h and 15-min exposure limits (threshold concentrations) of NH3 were exceeded in the broiler barn; the highest additive level was more than two times of the limit. Seasonal average emissions of NH3, H2S, and respirable dust were 57 g d-1 AU-1, 1.35 g d-1 AU-1, and 1.99 g d-1 AU-1, respectively, for the layer barn, all with higher levels in the mild and warm seasons than in the cold season. The emission data were only obtained for the worst-case scenarios (last week of the production cycle) of the broiler barn, with annual averages of 92 g d-1 AU-1 for NH3, 1.19 g d-1 AU-1 for H2S, and 4.32 g d-1 AU-1 for respirable dust, with obvious higher NH3 levels in winter. Additionally, manure removal once every 3-4 days for the layer barn reduced NH3 emissions by 62% and 90% in the cold and mild seasons, respectively. This study also found significant negative influence of outdoor T (Tout) on NH3 emissions for the broiler barn but positive impact of Tout on NH3 emissions for the layer barn.

Biogas production estimation using data-driven approaches for cold region municipal wastewater anaerobic digestion.

The objective of this study was to estimate biogas (including methane, carbon dioxide and hydrogen sulphide) production rates from the anaerobic digesters at the Saskatoon Wastewater Treatment Plant (SWTP), Saskatchewan, Canada. Average daily ambient temperatures typically fluctuate between -40 °C and 30 °C over the year making the management of the SWTP processes challenging. Operating parameters were taken from 2014 to 2016 including volatile fatty acids (VFAs), total solids, fixed solids, volatile solids, pH, and inflow rate. The input parameters were processed using two methods including a correlation test and principal component analysis (PCA) to determine highly correlated variables prior to use in models. The two models used to estimate biogas production rates are a multi-layered perceptron feed forward artificial neural network (ANN) and an adaptive network-based fuzzy inference system (ANFIS) with grid partition (GP), subtractive clustering (SC) and fuzzy c-means clustering (FCMC). The models using PCA processed variables had reasonable performances with shorter model processing times, while reducing model input data. Among various structures of ANN and ANFIS models for estimation of biogas generation, the ANFIS-FCMC results had better agreement with the observed data. Its average approximation of emission rates of CH4, CO2 and H2S from the wastewater digesters were 3,086, 6,351, and 41.5 g/min, respectively. Our group is assessing similar estimation methodology for the remaining SWTP wastewater treatment processes that are more highly impacted by the seasonal temperature variations including primary and secondary treatment processes.

Dispersion modeling of odour, gases, and respirable dust using AERMOD for poultry and dairy barns in the Canadian Prairies.

For determining setback distances considering multiple air pollutants, a comprehensive study was conducted to simulate the atmospheric dispersion of odour, ammonia (NH3), hydrogen sulfide (H2S), and respirable dust using an US EPA air dispersion model AERMOD for a commercial dairy, broiler, and cage-layer barn in the Canadian Prairies. The simulation was conducted using five years of meteorological data. Setback distances were determined with the input of varying monthly emission rates of all four air pollutants and odour impact criteria specifically developed for all three odour sources. Results showed the layer barn had the greatest odour impact area (maximum 3023 m for an annual average odour concentration of 0.01 OU m-3) followed by the broiler and dairy barns. Due to the prevailing south wind for all three barns, odour traveled farthest in the north. Using the suggested odour impact criteria by the Government of Saskatchewan defined for all odour sources, maximum setback distances were decreasing from 1941 to 641 m for the layer barn and from 980 to 320 m for the broiler barn along with the increasing of odour concentration (OC) thresholds (1-6 OU m-3), all in the north direction. While for the dairy barn, setback distances were determined only under an OC limit of 1 OU m-3; maximum 205 m in the north and minimum 171 m in the south. Using the newly developed odour impact criteria specifically for the three odour sources, maximum setback distance of 558 m in the north was determined for the layer barn under an odour threshold of 9 OU m-3. Additionally, the results suggest the use of odour impact criteria for determining setback distance rather than using gas/respirable threshold limits set in ambient air quality standards as the former always requires much greater setback distances than the latter.

Diurnal and seasonal variations of greenhouse gas emissions from a commercial broiler barn and cage-layer barn in the Canadian Prairies.

Baseline emission values of greenhouse gases were not well established for commercial poultry barns in cold regions, including Canada, due to a lack of well-designed field studies. Emission factors of carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O), were acquired for a commercial broiler barn and cage-layer barn in the Canadian Prairies climate. Between March 2015 and February 2016, monthly measurements throughout the year for the layer barn and over 6 flocks for the broiler barn, and diurnal measurements in the mild, warm, and cold seasons for both barns were conducted, respectively. The ventilation rate was estimated based on a CO2 mass balance method; thus CO2 emissions were quantified by the CIGR (2002) models. The CH4 and N2O emissions present at low levels from global perspective for both barns; the cold climate proved to be a major reason for the lower CH4 emission from the layer barn. Considerable seasonal effect was observed only for N2O emissions from the broiler barn, and for CH4 and N2O emissions from the layer barn, both with higher emissions in the mild and warm seasons than in the cold season. The big diurnal variations of CO2 emissions for the layer barn demonstrated the uncertainty of the seasonal results by snapshot measurements and correction factors (from -20.9% to -22.5%) were obtained. Besides, the difference of CH4 and N2O concentrations and emissions as well as CO2 concentrations between best-case (the first day after manure removal) and worst-case conditions (the last day before manure removal) was not obvious for the layer barn. Additionally, changes of temperature and ventilation rate were likely to have more impact on N2O emission for the broiler barn and more impact on CH4 emission for the layer barn than on the other two gas emissions, both with positive correlations.

Synthesis, characterization and activity performance of nickel-loaded spent FCC catalyst for pine gum hydrogenation.

A Ni-based catalyst supported over a spent fluid catalytic cracking (FCC) catalyst was prepared by a wet impregnation method. The catalytic characteristic was investigated in pine gum hydrogenation. Optimum conditions for catalyst preparation were obtained as: 15% Ni loading, 723 K calcination temperature, and 723 K reduction temperature for 2.5 h. The characterization results indicate that the specific surface area of the spent catalyst increased from 65.70 m2 g-1 to 67.78 m2 g-1 after calcination. The H2-TPR profile of the spent FCC catalyst exhibited two reduction peaks at 1123 °C and 670 °C. The TG curves showed that the second and third steps occur at 440 K and 550 K, respectively, having a total weight loss of 16%. The NiO grains possess rhombus particles on the surface or between the layers of the supported NiO catalyst. After activation in H2 flow, the metallic Ni loads on the support with no covalent bond between them. The NH3-TPD results indicated that the spent FCC catalyst held an obvious distribution of weakly acidic sites, and the acid sites became stronger after loading the catalyst with Ni. The hydrogenation products of pine gum were identified by GC-MS and a reaction mechanism was proposed. This study demonstrated its cost-effectiveness, environment-friendly nature and that the utilization of a spent FCC catalyst can be effectively applied as an alternative approach to pine gum hydrogenation on an industrial level.

A polysaccharide from green tea (Camellia sinensis L.) protects human retinal endothelial cells against hydrogen peroxide-induced oxidative injury and apoptosis.

Oxidative damage of retinal pigment epithelium (RPE) cells is involved in the pathogenesis age related macular degeneration (AMD). The purpose of this study was to evaluate the potential protective effect of a purified green tea polysaccharide (GTWP) against hydrogen peroxide (H2O2) induced oxidative stress and apoptosis in human retinal pigment epithelial cells (ARPE-19 cells). Human ARPE-19 cells were treated with 1 h of 500 µM H2O2 before incubation with GTWP for 24 h. Pretreatment of GTWP decreased H2O2-induced cell death and cell apoptosis, and efficiently suppressed the intracellular ROS production and malondialdehyde (MDA) generation induced by H2O2 treatment. Moreover, a loss of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) activities were restored to normal level in H2O2-induced ARPE-19 cells upon GTWP (100 µg/ml) exposure. Also, the tendency of increased protein expression of Bax and cleaved-caspsae-3, as well as decrease of Bcl-2 protein in ARPE-19 cells challenged with H2O2 was changed to individual opposite way, thus inhibiting the apoptotic cell death. Our results demonstrated that GTWP protected RPE cells against oxidative injury through activation of anti-apoptotic and endogenous antioxidant enzymes signaling pathway, suggesting GTWP has attractive therapeutic potential to AMD.

Relationships between odor properties and determination of odor concentration limits in odor impact criteria for poultry and dairy barns.

Livestock odor properties have not been well understood and the role of hedonic tone (HT) in establishing appropriate odor impact criteria has not been investigated. Five odor properties, including odor concentration (OC), intensity (OI), HT, persistence, and character descriptor, were studied for odorous air from a commercial dairy barn, layer barn, and broiler barn by taking measurements in all four seasons. The seasonal OC of the dairy, layer, and broiler barns averaged 447±162OUm-3, 583±216OUm-3, and 766±148OUm-3, respectively. Correspondingly, OI and HT averaged 2.7±0.5 and -2.6±0.5 for the dairy barn, 2.9±0.4 and -2.9±0.5 for the layer barn, and 3.2±0.4 and -3.1±0.4 for the broiler barn. Significant correlations were observed among OC, OI, and HT for all three odors (P<0.01). Increased OC was accompanied by increased OI but decreased HT. The relationships between OC and OI, and between OC and HT were derived in both Weber-Fechner law and Stevens' power law, while the best relationship between OI and HT turned out to be in a cubic polynomial model for the dairy-barn odor and a quadratic polynomial model for the two poultry barn odors. Based on OI-OC and HT-OC relationships in Weber-Fechner law, a reference table of OC limits was generated with 3 set values for OI (0, 1, and 2) and HT (0, -1, and -2) for all three odor sources, which may provide references in establishing appropriate odor impact criteria to meet different land use purposes. The comparison of the OC limits was made using relationships for all odor samples and for odor below 320OUm-3 (OI=3), indicating no significant difference. Slightly lower OCs from the former were suggested for use in stricter odor impact criteria.

[Metabolic fingerprint analysis of RAW264.7 inflammatory cell model by using UPLC-Q-TOF/MS].

In order to reveal the properties of polar metabolome in inflammatory cells, we selected LPS-induced RAW264.7 inflammatory cell models as the carrier for the research of metabolic fingerprint analysis. In this study, an ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics protocol was optimized for the extraction of polar metabolites from RAW264.7 cell line. Then orthogonal partial least squares discriminant analysis (OPLS-DA) was used to process the metabolic data, and finally, a total of 17 metabolites were selected and identified. The results showed that MeOH-CHCl3-H2O (8∶1∶1) was chosen as the optimal extraction solvent to achieve higher number of chromatographic peaks, with the best relative extraction efficiency and stability. Comparing with the normal cells, the inflammatory cells presented an abnormal metabolism in protein, carbohydrate, nucleotide and phospholipids. In this study, a UPLC-Q-TOF/MS-based metabolomics protocol for the polar metabolites from RAW264.7 cell line was developed, which may provide important information for the study of mechanism of inflammation and the anti-inflammatory drugs.

Diurnal and seasonal variations of odor and gas emissions from a naturally ventilated free-stall dairy barn on the Canadian prairies.

This study characterized the seasonal concentration (C) and emission (E) patterns of odor, ammonia (NH3), and hydrogen sulfide (H2S) over the course of a whole year and their diurnal patterns in cold, warm, and mild seasons for a naturally ventilated free-stall dairy barn. It was found that seasonal odor and NH3 and H2S emissions varied greatly: from 17.2 to 84.4 odor units (OU) sec-1 AU-1 (AU: animal unit, 500 kg of animal body mass), from 0.27 to 0.92 mg sec-1 AU-1, and from 3 to 105 µg sec-1 AU-1, respectively. The overall concentrations of odor and NH3 were higher in the winter, whereas the emissions were higher in the mild and warm seasons. Diurnal variation was most significant for odor emission (OE) in the mild season when the ratio of maximum (279.2 OU sec-1 AU-1) to minimum value (60.5 OU sec-1 AU-1) was up to 4.6. The indoor air quality was also evaluated by considering not only the health effect of individual gases, but also the additive effect of NH3 and H2S. Results showed that the indoor air quality was poorest in cold seasons when NH3 C could exceed the threshold limit set out in occupational health regulation, and in fact could worsen due to the additive effect of the two gases. Further, it was suggested NH3 was a good indicator for predicting odor concentration (OC) or OE. The impact of climatic parameters on odor and gases were also examined, and it was found ventilation rate (VR) negatively affected OC and NH3 C, but positively impacted OE and NH3 E. Using 70% of the total data, a multilinear model for OE was developed as a function of VR and indoor relative humidity and was validated to be acceptable using the rest of the data. IMPLICATIONS: Diurnal and seasonal variations of odor, NH3, and H2S concentrations and emissions were monitored for a naturally ventilated dairy barn in a cold region. The emission factors were calculated and indoor air quality was evaluated. The overall odor and NH3 concentrations were higher in winter, whereas emissions were higher in the mild and warm seasons. Diurnal variation was most significant for odor emission in the mild season, when the ratio of maximum to minimum value was up to 4.6. The results can be used to estimate odor and gas emissions from other dairy barns in Canada and other cold regions.

A new spot quality control for protein macroarray based on immunological detection.

Protein macroarray is a new, simple and multiple biochemistry detection system, in which the test spots are more than 1mm diameter and results directly visible and instrumentation is not necessary. This technology, however, possesses recognized problems with spot quality and uniformity, issues that can limit its application. Previous methods have been developed for spot quality control, but they are complicated or require specific instrumentation. Therefore, we have developed a spot quality control buffer supplement with Ponceau S (SQCB-PS) as a direct and visible means of monitoring spot quality on nitrocellulose membrane prior to hybridization. In this report, 1% (w/v) Ponceau S and 10% (v/v) glycerol in spotting buffer were found to be optimal for spot quality control and food-borne pathogens multiple detection. Furthermore, the sensitivity and specificity of the protein macroarray were not compromised by spotting buffer with Ponceau S. Under optimal conditions, this visible spot quality control method makes the detection more reliable, and should enhance the wider use of the technique.

The molecular mechanisms of interactions between bioactive peptides and angiotensin-converting enzyme.

The ability of milk protein derived Ile-Pro-Ala (IPA), Phe-Pro (FP) and Gly-Lys-Pro (GKP) peptides to inhibit angiotensin I-converting enzyme (ACE), a protein with an important role in blood-pressure regulation, were verified in vitro and in vivo. This work elucidates the modes and molecular mechanisms of the interaction of IPA, FP and GKP with ACE, including mechanisms that bind the peptides to the cofactor Zn(2+). It was observed that the best docking poses obtained for IPA, FP and GKP were at the ACE catalytic site with very similar modes of interaction, including the interaction with Zn(2+). The interactions, including H-bonds, hydrophobic, hydrophilic, and electrostatic interactions, as well as the interaction with Zn(2+), were responsible for the binding between the bioactive peptides and ACE.

Development of a livestock odor dispersion model: part I. Model theory and development.

A livestock odor dispersion model (LODM) was developed to predict mean odor concentration, odor frequency, instantaneous odor concentration, and peak odor concentration from livestock operations. This model is based on the Gaussian fluctuating plume model and has the ability to consider the instantaneous concentration fluctuations and the differences between odor and traditional air pollutants. It can predict odor frequency from the routine hourly meteorological data input and deal with different types of sources and multiple sources. Also, the relationship between odor intensity and odor concentration was incorporated into the model.

Development of a livestock odor dispersion model: part II. Evaluation and validation.

A livestock odor dispersion model (LODM) was developed to predict odor concentration and odor frequency using routine hourly meteorological data input. The odor concentrations predicted by the LODM were compared with the results obtained from other commercial models (Industrial Source Complex Short-Term model, version 3, CALPUFF) to evaluate its appropriateness. Two sets of field odor plume measurement data were used to validate the model. The model-predicted mean odor concentrations and odor frequencies were compared with those measured. Results show that this model has good performance for predicting odor concentrations and odor frequencies.

Annual variations of odor concentrations and emissions from swine gestation, farrowing, and nursery buildings.

To obtain annual odor emission profiles from intensive swine operations, odor concentrations and emission rates were measured monthly from swine nursery, farrowing, and gestation rooms for a year. Large annual variations in odor concentrations and emissions were found in all the rooms and the impact of the seasonal factor (month) was significant (P < 0.05). Odor concentration was low in summer when ventilation rate was high but high in winter when ventilation rate was low, ranging from 362 (farrowing room in July) to 8934 (nursery room in December) olfactory unit (OU) m(-3). This indicates that the air quality regarding odor was significantly better in summer than that in winter. Odor emission rate did not show obvious seasonal pattern as odor concentration did, ranging from 2 (gestation room in November) to 90 (nursery room in April) OU m(-2) sec(-1); this explains why the odor complaints for swine barns have occurred all year round. The annual geometric mean odor concentration and emission rate of the nursery room was significantly higher than the other rooms (P < 0.05). In order to obtain the representative annual emission rate, measurements have to be taken at least monthly, and then the geometric mean of the monthly values will represent the annual emission rate. Incorporating odor control technologies in the nursery area will be the most efficient in reducing odor emission from the farm considering its emission rate was 2 to 3 times of the other areas. The swine grower-finisher area was the major odor source contributing 53% of odor emission of the farm and should also be targeted for odor control. Relatively positive correlations between odor concentration and both H2S and CO2 concentrations (R(2) = 0.58) means that high level of these two gases might likely indicate high odor concentration in swine barns.

Determination of setback distances for livestock operations using a new livestock odor dispersion model (LODM).

Setback distance has been used as an effective tool to avoid odor nuisance from livestock operations. Many setback distances were guidelines that were determined by empirical methods that are considered to be lack of science base. Air dispersion models have been used to determine setback distances; however, these models do not consider the short-time fluctuations of odor. A livestock odor dispersion model (LODM) was developed to consider the short-time variations of odor and predict occurrence frequency for certain levels of odor. In this study, this model was used to predict the occurrence frequency for various levels of odor in the vicinity (10 km) of a swine farm. Using selected odor criteria, setback distances between the swine farm and nearby communities were defined. Results indicate that the LODM can be used as an effective tool to determine setback distances.

Seasonal odor, ammonia, hydrogen sulfide, and carbon dioxide concentrations and emissions from swine grower-finisher rooms.

Seasonal odor and gas (ammonia [NH3], hydrogen sulfide [H2S], and carbon dioxide [CO2]) concentrations and emission rates (OGCERs) from swine facilities are vital for providing accurate source emissions and reducing the uncertainty of setback distances on the basis of emission data. In this study, a repeated measurement experimental method and a split-block statistical model were used to obtain seasonal OGCER profiles from two types of swine grower-finisher rooms in Saskatchewan, Canada, over a 12-month period. The results indicate that the OGCERs were significantly affected by the sampling month and ambient temperature (P < 0.05), which indicates that monthly OGCERs should be measured and used as representative monthly or seasonal values in air dispersion models to reduce uncertainties in setback calculations. It was also found that the seasonal OGCERs from the rooms with fully slatted floors were 6.3-40.6% higher than those with partially slatted floors. The seasonal OGCERs (except for the NH3 concentrations in October, November, and January; the CO2 concentrations in August; and the CO2 emission rates in December) between these two rooms for each measuring month did not differ significantly (P > 0.05). The measured gas concentrations were generally below the permissible exposure limits (PELs) established by the Occupational Safety and Health Administration (OSHA) throughout the year except for the NH3 concentrations in cold weather (December, January, and February).

Diurnal odor, ammonia, hydrogen sulfide, and carbon dioxide emission profiles of confined swine grower/finisher rooms.

The objective of this study was to obtain diurnal variation profiles of odor and gas (ammonia [NH3], hydrogen sulfide [H2S], carbon dioxide [CO2]) concentrations and emission rate (OGCER) from confined swine grower/ finisher rooms under three typical weather conditions (warm, mild, and cold weather) in a year. Two grower/ finisher rooms, one with a fully slatted floor and the other with partially slatted floors, were measured for 2 consecutive days under each weather condition. The results revealed that the diurnal OGCER in the room with a fully slatted floor was 9.2-39.4% higher than that with a partially slatted floor; however, no significant differences in the diurnal OGCER were found between these two rooms, except for the NH3 concentrations in August, the NH3 and H2S concentrations and emissions in October, and odor concentrations and emissions in February (p > 0.05). The OGCER variations presented different diurnal patterns as affected by time of day, season, type of floor, ventilation rate, animal growth cycles, in-house manure storage, and weather conditions. Significant diurnal fluctuations in the OGCER (except for the odor concentrations and H2S emissions) were observed in August (p < 0.05); all of the gas emissions in October and the CO2 concentrations and emissions in February also showed significant diurnal variations (p < 0.05). These significant diurnal variations indicated that the OGCER during different periods of a day should be monitored when quantifying OGCER concentrations and emissions; for example, source emission data used in air dispersion modeling to decrease the great incertitude of setback determination using randomly measured data.