Tracking and Characterizing Spatiotemporal and Three-Dimensional Locomotive Behaviors of Individual Broilers in the Three-Point Gait-Scoring System.

Gait scoring is a useful measure for evaluating broiler production efficiency, welfare status, bone quality, and physiology. The research objective was to track and characterize spatiotemporal and three-dimensional locomotive behaviors of individual broilers with known gait scores by jointly using deep-learning algorithms, depth sensing, and image processing. Ross 708 broilers were placed on a platform specifically designed for gait scoring and manually categorized into one of three numerical scores. Normal and depth cameras were installed on the ceiling to capture top-view videos and images. Four birds from each of the three gait-score categories were randomly selected out of 70 total birds scored for video analysis. Bird moving trajectories and 16 locomotive-behavior metrics were extracted and analyzed via the developed deep-learning models. The trained model gained 100% accuracy and 3.62 ± 2.71 mm root-mean-square error for tracking and estimating a key point on the broiler back, indicating precise recognition performance. Broilers with lower gait scores (less difficulty walking) exhibited more obvious lateral body oscillation patterns, moved significantly or numerically faster, and covered more distance in each movement event than those with higher gait scores. In conclusion, the proposed method had acceptable performance for tracking broilers and was found to be a useful tool for characterizing individual broiler gait scores by differentiating between selected spatiotemporal and three-dimensional locomotive behaviors.

Perches as Cooling Devices for Reducing Heat Stress in Caged Laying Hens: A Review.

Heat stress is one of the most detrimental environmental challenges affecting the biological process and the related production performance of farm animals, especially in poultry. Commercial laying hens have been bred (selected) for high egg production, resulting in increased sensitivity to heat stress due to breeding-linked metabolic heat production. In addition, laying hens are prone to heat stress due to their inadequate species-specific cooling mechanisms resulting in low heat tolerance. In addition, hens have no sweat glands and feathering covers almost their entire body to minimize body heat loss. The poultry industry and scientists are developing cooling methods to prevent or reduce heat stress-caused damage to chicken health, welfare, and economic losses. We have designed and tested a cooling system using perches, in which chilled water (10 °C) circulates through a conventional perch passing through the layer cages to offer the cooling potential to improve hen health, welfare, and performance during acute and chronic periods of heat stress (35 °C). This review summarizes the outcomes of a multi-year study using the designed cooled perch system. The results indicate that conducting heat from perching hens directly onto the cooled perch system efficiently reduces heat stress and related damage in laying hens. It provides a novel strategy: perches, one key furnishment in cage-free and enriched colony facilities, could be modified as cooling devices to improve thermal comfort for hens during hot seasons, especially in the tropical and subtropical regions.

Microbial assemblages and methanogenesis pathways impact methane production and foaming in manure deep-pit storages.

Foam accumulation in swine manure deep-pits has been linked to explosions and flash fires that pose devastating threats to humans and livestock. It is clear that methane accumulation within these pits is the fuel for the fire; it is not understood what microbial drivers cause the accumulation and stabilization of methane. Here, we conducted a 13-month field study to survey the physical, chemical, and biological changes of pit-manure across 46 farms in Iowa. Our results showed that an increased methane production rate was associated with less digestible feed ingredients, suggesting that diet influences the storage pit's microbiome. Targeted sequencing of the bacterial 16S rRNA and archaeal mcrA genes was used to identify microbial communities' role and influence. We found that microbial communities in foaming and non-foaming manure were significantly different, and that the bacterial communities of foaming manure were more stable than those of non-foaming manure. Foaming manure methanogen communities were enriched with uncharacterized methanogens whose presence strongly correlated with high methane production rates. We also observed strong correlations between feed ration, manure characteristics, and the relative abundance of specific taxa, suggesting that manure foaming is linked to microbial community assemblage driven by efficient free long-chain fatty acid degradation by hydrogenotrophic methanogenesis.

Classifying Ingestive Behavior of Dairy Cows via Automatic Sound Recognition.

Determining ingestive behaviors of dairy cows is critical to evaluate their productivity and health status. The objectives of this research were to (1) develop the relationship between forage species/heights and sound characteristics of three different ingestive behaviors (bites, chews, and chew-bites); (2) comparatively evaluate three deep learning models and optimization strategies for classifying the three behaviors; and (3) examine the ability of deep learning modeling for classifying the three ingestive behaviors under various forage characteristics. The results show that the amplitude and duration of the bite, chew, and chew-bite sounds were mostly larger for tall forages (tall fescue and alfalfa) compared to their counterparts. The long short-term memory network using a filtered dataset with balanced duration and imbalanced audio files offered better performance than its counterparts. The best classification performance was over 0.93, and the best and poorest performance difference was 0.4-0.5 under different forage species and heights. In conclusion, the deep learning technique could classify the dairy cow ingestive behaviors but was unable to differentiate between them under some forage characteristics using acoustic signals. Thus, while the developed tool is useful to support precision dairy cow management, it requires further improvement.

Effects of drying and providing supplemental oxygen to piglets at birth on rectal temperature over the first 24 h after birth.

Neonatal piglets can experience both a decrease in body temperature and hypoxia, increasing risks for pre-weaning mortality. This research evaluated the effects of drying and providing supplemental oxygen to newborn piglets on rectal temperature (RT) over the first 24 h after birth. The study used a CRD with three Intervention Treatments (IT; applied at birth): Control (no intervention), Drying (dried using a desiccant), Oxygen [dried using a desiccant and placed in a chamber (at 40% oxygen concentration) for 20 min]. A total of 42 litters (485 piglets) were randomly allotted to treatments at the start of farrowing. At birth, each piglet was given a numbered ear tag, weighed, and the treatment was applied; RT was measured at 0, 20, 30, 45, 60, 120, and 1440 min after birth. Blood was collected from one piglet from each birth weight quartile within each litter at 24 h after birth to measure plasma immunocrit concentration. There was no effect (P > 0.05) of IT on piglet RT at 0 or 1440 min after birth. Between 20 and 60 min after birth, piglet RT was lower (P ≤ 0.05) for the Control than the Drying treatment, with the Oxygen treatment being intermediate and different (P ≤ 0.05) from the other two IT. The effect of piglet birth weight on responses to IT were evaluated by classifying piglets into Birth Weight Categories (BWC): Light (<1.0 kg), Medium (1.0 to 1.5 kg), or Heavy (>1.5 kg). There were IT by BWC interactions (P ≤ 0.05) for piglet RT at all measurement times between 20 and 120 min after birth. Relative to the Control, the effects of the Drying and Oxygen treatments on RT were greater (P ≤ 0.05) for Light than heavier piglets. Plasma immunocrit concentrations tended (P = 0.07) to be greater for piglets on the Control treatment compared to the other two IT and were lower (P ≤ 0.05) for Light than Heavy piglets, with Medium piglets being intermediate and different (P ≤ 0.05) to the other BWC. In conclusion, drying piglets at birth reduced the extent and duration of RT decline in piglets in the early postnatal period compared to undried piglets, especially for those of low birth weight. However, the combination of drying and placing piglets in an oxygen-rich environment provided no additional benefit over drying alone.

Testing the plastic-wrapped composting system to dispose of swine mortalities during an animal disease outbreak.

Composting has been used to dispose of animal mortalities and infected materials, such as manure and feed, during major animal disease outbreaks. In this study, we adapted the plastic-wrapped mortality composting system developed by the Canadian Food Inspection Agency during the 2004 highly pathogenic avian influenza outbreak to compost swine mortalities. The goals of the study were to evaluate the performance of the plastic-wrapped composting system to dispose of swine mortalities and to field test its ability to eliminate the spread of airborne pathogens through the aeration ducts. Two cover materials, ground cornstalks and woodchips, were tested using passively and actively aerated composting sheds. The mortalities were inoculated with Salmonella spp. and vaccine strains of Bovine herpesvirus-1 and Bovine viral diarrhea virus. Air samples collected from the upper aeration duct (air outlet) during the first 10 d of composting were negative for Salmonella and the viruses tested, which indicated that aerosol transmission of the pathogens was limited. The aeration plenum placed under the mortalities helped to keep conditions aerobic, as O2 concentrations of both passively and actively aerated test units were above 11%. Actively aerated cornstalks had the highest degree-hours (1,462 °C h d-1 ), which was followed by passively aerated cornstalks (1,312 °C h d-1 ), actively aerated woodchips (1,303 °C h d-1 ), and passively aerated woodchips (1,062 °C h d-1 ). After a 7-wk composting period, all three pathogens were inactivated based on quantitative polymerase chain reaction test results. The mortalities were not inoculated with the African swine fever virus, but temperature data showed that if they were, the system had the potential to eliminate this virus.

Wireless 'under the skull' epidural EEG and behavior in piglets during nitrous oxide or carbon dioxide gas euthanasia.

Consciousness is central to animal welfare concerns. Its assessment is most often conducted based on behavior, with a poor understanding of the correspondence between behavior and the neurobiological processes that underlie the subjective experience of consciousness. Recording of brain electrical activity using electrodes placed under the skull improves EEG recording by minimizing artifacts from muscular or cardiac activities, and it can now be combined with wireless recording in free-moving animals. This experiment investigated the correspondence between wireless 'under the skull' epidural EEG and the behavior of 18 five-week-old female piglets undergoing nitrous oxide (N2O) or carbon dioxide (CO2) gradual fill gas euthanasia at 25% replacement rate per minute of the chamber volume. Piglets exposed to CO2 had a peak in EEG total power ('Ptot') during the flailing stage, whereas piglets exposed to N2O had a higher EEG 95% spectral edge frequency ('F95') during their initial explorative behavior phase and a drop in EEG median frequency ('F50') after loss of posture. Loss of posture without righting attempt, as the last behavioral state observed during euthanasia, preceded the onset of transitional EEG on average by 0.9 and 3.1 min (for CO2 and N2O treatments, respectively), and the onset of isoelectric EEG by 4.5 and 6.2 min (for CO2 and N2O treatments, respectively). Paddling movements occurred shortly before and during transitional EEG but never during isoelectric EEG, whereas gasps persisted after the EEG had become isoelectric. The dynamics of EEG spectral changes were complex to interpret in relation to the degree of consciousness, but isoelectric EEG as an unequivocal indicator of unconsciousness appeared several minutes after loss of posture with no righting attempt. This leaves a window of uncertainty in regards to the potential for consciousness after loss of posture during gradual fill gas euthanasia in piglets.

Pig barns ammonia and greenhouse gas emission mitigation by slurry aeration and acid scrubber.

Livestock production is associated with several gaseous pollutant emissions to the environment. These emissions can degrade local and regional air quality, contribute to surface water eutrophication and acid rain, and contribute to the greenhouse gas footprint of the production sector. Modern production systems must balance animal welfare and environmental pollution potential with economic reality, which is a great challenge to maintain as global demand for animal protein increases. Accordingly, gaseous emission technologies were the main target for this research, in which mitigating gas emissions of ammonia, nitrous oxide, and methane from pig production facilities via slurry aeration system was tested. Five treatments with different airflow rates in the test room were examined continuously over a period of 6 weeks and the results were compared with the control room. Test results indicate that the highest mitigation potentials were 12, 57.6, and 10.4% for nitrous oxide, methane, and ammonia, respectively. Subsequently directing exhaust air into a sulfuric acid air scrubber at 3.0 pH further reduced total ammonia emissions by 80 to 87%.

Factors Affecting Trailer Thermal Environment Experienced by Market Pigs Transported in the US.

Extreme weather conditions challenge pig thermoregulation during transport and are addressed by the National Pork Board (NPB) Transport Quality Assurance® (TQA) program that provides guidelines for trailer boarding, bedding, and misting. These guidelines are widely applied, yet very little is known about the microenvironment within the trailer. In this study, TQA guidelines (V4) were evaluated via extensive thermal environment measurements during transport in order to evaluate spatial variability and implications on ventilation pattern. Effects of trailer management strategies including bedding, boarding, and misting were examined and the trailer was monitored for interior temperature rise and THI responses within six separate zones. The trailer thermal environment was not uniformly distributed in the colder trips with the top front and bottom zones were the warmest, indicating these zones had the majority of outlet openings and experienced air with accumulated sensible and latent heat of the pigs. Relatively enhanced thermal environment uniformity was observed during hot trips, suggesting that ventilation patterns and ventilation rate were different for colder vs. warmer weather conditions. Misting applied prior to transport cooled interior air temperature, but also created high THI conditions in some cases. Neither boarding and bedding combinations in the TQA nor boarding position showed impacts on trailer interior temperature rise or spatial distribution of temperature inside the trailer.

A Two-Step Process of Nitrous Oxide before Carbon Dioxide for Humanely Euthanizing Piglets: On-Farm Trials.

Current methods of euthanizing piglets are raising animal welfare concerns. Our experiment used a novel two-step euthanasia method, using nitrous oxide (N2O) for six minutes and then carbon dioxide (CO2) on compromised 0- to 7-day-old piglets. A commercial euthanasia chamber was modified to deliver two euthanasia treatments: the two-step method using N2O then CO2 (N2O treatment) or only CO2 (CO2 treatment). In Experiment 1, 18 piglets were individually euthanized. In Experiment 2, 18 groups of four to six piglets were euthanized. In the N2O treatment, piglets lost posture, indicating the onset of losing consciousness, before going into CO2 where they showed heavy breathing and open-mouth breathing; whereas piglets in the CO2 treatment did not lose posture until after exhibiting these behaviors (p ≤ 0.004). However, piglets in the N2O treatment took longer to lose posture compared to the CO2 treatment (p < 0.001). Piglets in the N2O treatment displayed more behavioral signs of stress and aversion: squeals/minute (p = 0.004), escape attempts per pig (p = 0.021), and righting responses per pig (p = 0.084) in a group setting. In these regards, it cannot be concluded that euthanizing piglets for 6 min with N2O and then CO2 is more humane than euthanizing with CO2 alone.

Relationships between chemical structure, mechanical properties and materials processing in nanopatterned organosilicate fins.

The exploitation of nanoscale size effects to create new nanostructured materials necessitates the development of an understanding of relationships between molecular structure, physical properties and material processing at the nanoscale. Numerous metrologies capable of thermal, mechanical, and electrical characterization at the nanoscale have been demonstrated over the past two decades. However, the ability to perform nanoscale molecular/chemical structure characterization has only been recently demonstrated with the advent of atomic-force-microscopy-based infrared spectroscopy (AFM-IR) and related techniques. Therefore, we have combined measurements of chemical structures with AFM-IR and of mechanical properties with contact resonance AFM (CR-AFM) to investigate the fabrication of 20-500 nm wide fin structures in a nanoporous organosilicate material. We show that by combining these two techniques, one can clearly observe variations of chemical structure and mechanical properties that correlate with the fabrication process and the feature size of the organosilicate fins. Specifically, we have observed an inverse correlation between the concentration of terminal organic groups and the stiffness of nanopatterned organosilicate fins. The selective removal of the organic component during etching results in a stiffness increase and reinsertion via chemical silylation results in a stiffness decrease. Examination of this effect as a function of fin width indicates that the loss of terminal organic groups and stiffness increase occur primarily at the exposed surfaces of the fins over a length scale of 10-20 nm. While the observed structure-property relationships are specific to organosilicates, we believe the combined demonstration of AFM-IR with CR-AFM should pave the way for a similar nanoscale characterization of other materials where the understanding of such relationships is essential.

Experimental confirmation of the atomic force microscope cantilever stiffness tilt correction.

The tilt angle (angle of repose) of an AFM cantilever relative to the surface it is interrogating affects the effective stiffness of the cantilever as it analyzes the surface. For typical AFMs and cantilevers that incline from 10° to 15° tilt, this is thought to be a 3%-7% stiffness increase correction. While the theoretical geometric analysis of this effect may have reached a consensus that it varies with cos-2 θ, there is very little experimental evidence to confirm this using AFM cantilevers. Recently, the laser Doppler vibrometry thermal calibration method utilized at NIST has demonstrated sufficient stiffness calibration accuracy, and precision to allow a definitive experimental confirmation of the particular trigonometric form of this tilt effect using a commercial microfabricated AFM cantilever specially modified to allow strongly tilted (up to 15°) effective cantilever stiffness measurements.

Characteristics of Trailer Thermal Environment during Commercial Swine Transport Managed under U.S. Industry Guidelines.

Transport is a critical factor in modern pork production and can seriously affect swine welfare. While previous research has explored thermal conditions during transport, the impact of extreme weather conditions on the trailer thermal environment under industry practices has not been well documented; and the critical factors impacting microclimate are not well understood. To assess the trailer microclimate during transport events, an instrumentation system was designed and installed at the central ceiling level, pig level and floor-level in each of six zones inside a commercial swine trailer. Transport environmental data from 34 monitoring trips (approximately 1-4 h in duration each) were collected from May, 2012, to February, 2013, with trailer management corresponding to the National Pork Board Transport Quality Assurance (TQA) guidelines in 31 of these trips. According to the TQA guidelines, for outdoor temperature ranging from 5 °C (40 °F) to 27 °C (80 °F), acceptable thermal conditions were observed based on the criteria that no more than 10% of the trip duration was above 35 °C (95 °F) or below 0 °C (32 °F). Recommended bedding, boarding and water application were sufficient in this range. Measurements support relaxing boarding guidelines for moderate outdoor conditions, as this did not result in less desirable conditions. Pigs experienced extended undesirable thermal conditions for outdoor temperatures above 27 °C (80 °F) or below 5 °C (40 °F), meriting a recommendation for further assessment of bedding, boarding and water application guidelines for extreme outdoor temperatures. An Emergency Livestock Weather Safety Index (LWSI) condition was observed inside the trailer when outdoor temperature exceeded 10 °C (50 °F); although the validity of LWSI to indicate heat stress for pigs during transport is not well established. Extreme pig surface temperatures in the rear and middle zones of the trailer were more frequently experienced than in the front zones, and the few observations of pigs dead or down upon arrival were noted in these zones. Observations indicate that arranging boarding placement may alter the ventilation patterns inside the trailer.

Accurate flexural spring constant calibration of colloid probe cantilevers using scanning laser Doppler vibrometry.

Calibration of the flexural spring constant for atomic force microscope (AFM) colloid probe cantilevers provides significant challenges. The presence of a large attached spherical added mass complicates many of the more common calibration techniques such as reference cantilever, Sader, and added mass. Even the most promising option, AFM thermal calibration, can encounter difficulties during the optical lever sensitivity measurement due to strong adhesion and friction between the sphere and a surface. This may cause buckling of the end of the cantilever and hysteresis in the approach-retract curves resulting in increased uncertainty in the calibration. Most recently, a laser Doppler vibrometry thermal method has been used to accurately calibrate the normal spring constant of a wide variety of tipped and tipless commercial cantilevers. This paper describes a variant of the technique, scanning laser Doppler vibrometry, optimized for colloid probe cantilevers and capable of spring constant calibration uncertainties near ±1%.

A simple methodology to measure ammonia flux generated in naturally ventilated poultry houses / Una metodología simple para medir el flujo de amoniaco generado en instalaciones avícolas sometidas a ventilación natural / Uma metodologia simples para quantificação do fluxo de amônia gerada em aviários submetidos à ventilação natural

Background: quantifying the rate of pollutant emissions, such as ammonia (NH3), from naturally ventilated animal production buildings in structures, such as those used in South America, is challenging mainly due to the challenge of determining building ventilation rate. Some methods have been developed to reduce this difficulty, but most are costly and complex, preventing their implementation.. Objective: the present work aimed to adapt and validate the Saraz method for the determination of ammonia emissions (SMDAE) from naturally ventilated poultry houses. Methods: to validate the proposed method, the ammonia emission rate (ER; NH3 mass emitted per unit time) was computed using the adjusted equation proposed by Wheeler et al.(2006; ER2). For evaluation purposes, the data obtained from the two measuring methods (SMDAE and ER2)were statistically analyzed. Results: the proposed method was considered adequate for use under conditions of natural ventilation with wind speeds greater than 0.1 m/s-1 and NH3 concentrations greater than 1 ppm. Conclusion: the method demonstrated a good correlation between its values and those obtained by the standard equation for calculating emissions based on NH3 concentration, air speed and temperature, which makes the Saraz method a reliable approach to develop ammonia inventories for naturally-ventilated poultry facilities.

Antecedentes: la cuantificación de la tasa de emision de contaminantes tales como el amoniaco (NH3) en instalaciones de producción animal con ventilación natural como las utilizadas en América del Sur es un reto, en parte debido a la dificultad para determinar la tasa de ventilación. Algunos métodos han sido desarrollados para reducir esta dificultad, pero la mayoría de ellos son costosos y complejos, lo que impide su aplicación. Objetivo: el presente trabajo tuvo como objetivo adaptar y validar el método Saraz para calcular el flujo de amoniaco (SMDAE) generado en galpones avícolas con ventilación natural. Métodos: para validar el método propuesto, el flujo de amoniaco (ER; masa de NH3 emitida por unidad de tiempo) fue comparado con los resultados obtenidos por medio de la ecuación ajustada propuesta por Wheeler et al. (2006; ER2). Para su evaluación, los valores obtenidos con los dos métodos (SMDAE y ER2) fueron comparados mediante análisis estadístico. Resultados: el método propuesto se considera adecuado para ser utilizado en las condiciones de ventilación natural con vientos superiores a 0,1 m/s-1 y concentraciones de NH3 superiores a 1 ppm. Conclusión: el método presentó una buena correlación entre los valores determinados y los obtenidos mediante la ecuación estandar para el cálculo de emisiones basada en la concentración de NH3, velocidad del aire y temperatura, lo que lo convierte en una alternativa confiable para hacer inventarios de amoníaco en instalaciones avícolas con ventilación natural.

Antecedentes: a quantificação da taxa de emissão de poluentes, tais como a amônia (NH3) em instalações animais abertas e com ventilação natural, como são usadas na América do Sul, é um desafio, em parte devido a dificuldade de determinação da taxa de ventilação. Assim, alguns métodos têm sido desenvolvidos para reduzir esta dificuldade, mas a maioria deles são onerosos e complexos, inviabilizando a sua aplicação. Objetivo: o presente trabalho objetivou-se adaptar e validar o método Saraz para determinar o fluxo de NH3 (SMDAE) emitida dos galpões avícolas submetidos à ventilação natural. Métodos: para fazer a validação do método proposto, o fluxo de amônia (ER; massa de NH3 gerada por unidade de tempo) foi comparada com os resultados obtidos por meio de equação ajustada proposta por Wheeler et al. (2006; ER2). Para a avaliação proposta, os valores obtidos com os dois métodos usados (SMDAE e ER2) foram comparados mediante analise estadística. Resultados: o método proposto mostrou poder ser usado com confiabilidade em condições de ventilação natural com ventos maiores que 0,1 m/s-1 e concentrações de NH3 maiores que 1 ppm. Conclusão:o método apresentou uma boa correlação entre os valores encontrados pelo método proposto e aqueles obtidos na equação característica para o cálculo de emissões a qual é baseada no conhecimento da concentração de NH3, velocidade e temperatura do ar, o que torna o método como uma alternativa viável para fazer inventários de amônia em instalações aviárias com ventilação natural.

Smart Electronic Laboratory Notebooks for the NIST Research Environment.

Laboratory notebooks have been a staple of scientific research for centuries for organizing and documenting ideas and experiments. Modern laboratories are increasingly reliant on electronic data collection and analysis, so it seems inevitable that the digital revolution should come to the ordinary laboratory notebook. The most important aspect of this transition is to make the shift as comfortable and intuitive as possible, so that the creative process that is the hallmark of scientific investigation and engineering achievement is maintained, and ideally enhanced. The smart electronic laboratory notebooks described in this paper represent a paradigm shift from the old pen and paper style notebooks and provide a host of powerful operational and documentation capabilities in an intuitive format that is available anywhere at any time.

Performance of an image analysis processing system for hen tracking in an environmental preference chamber.

Image processing systems have been widely used in monitoring livestock for many applications, including identification, tracking, behavior analysis, occupancy rates, and activity calculations. The primary goal of this work was to quantify image processing performance when monitoring laying hens by comparing length of stay in each compartment as detected by the image processing system with the actual occurrences registered by human observations. In this work, an image processing system was implemented and evaluated for use in an environmental animal preference chamber to detect hen navigation between 4 compartments of the chamber. One camera was installed above each compartment to produce top-view images of the whole compartment. An ellipse-fitting model was applied to captured images to detect whether the hen was present in a compartment. During a choice-test study, mean ± SD success detection rates of 95.9 ± 2.6% were achieved when considering total duration of compartment occupancy. These results suggest that the image processing system is currently suitable for determining the response measures for assessing environmental choices. Moreover, the image processing system offered a comprehensive analysis of occupancy while substantially reducing data processing time compared with the time-intensive alternative of manual video analysis. The above technique was used to monitor ammonia aversion in the chamber. As a preliminary pilot study, different levels of ammonia were applied to different compartments while hens were allowed to navigate between compartments. Using the automated monitor tool to assess occupancy, a negative trend of compartment occupancy with ammonia level was revealed, though further examination is needed.

Intermittent contact resonance atomic force microscopy.

The intermittent contact resonance atomic force microscopy (ICR-AFM) mode proposed here is a new frequency modulation technique performed in scanning force controlled AFM modes like force volume or peak force tapping. It consists of tracking the change in the resonance frequency of an eigenmode of a driven AFM cantilever during scanning as the AFM probe intermittently contacts a surface at a controlled applied maximum force (setpoint). A high speed data capture was used during individual oscillations to obtain detailed contact stiffness-force curve measurements on a two-phase polystyrene/poly(methyl methacrylate) film with sub-micrometer size domains. Through a suitable normalization, the measurements were analyzed by linear fits to provide an improved quantitative characterization of these materials in terms of their elastic moduli and adhesive properties.

Moisture effects on gas-phase biofilter ammonia removal efficiency, nitrous oxide generation, and microbial communities.

We established a four-biofilter setup to examine the effects of moisture content (MC) on biofilter performance, including NH3 removal and N2O generation. We hypothesized that MC increase can improve NH3 removal, stimulate N2O generation and alter the composition and function of microbial communities. We found that NH3 removal efficiency was greatly improved when MC increased from 35 to 55%, but further increasing MC to 63% did not help much; while N2O concentration was low at 35-55% MC, but dramatically increased at 63% MC. Decreasing MC from 63 to 55% restored N2O concentration. Examination of amoA communities using T-RFLP and real-time qPCR showed that the composition and abundance of ammonia oxidizers were not significantly changed in a "moisture disturbance-disturbance relief" process in which MC was increased from 55 to 63% and then reduced to 55%. This observation supported the changes of NH3 removal efficiency. The composition of nosZ community was altered at 63% MC and then was recovered at 55% MC, which indicates resilience to moisture disturbance. The abundance of nosZ community was negatively correlated with moisture content in this process, and the decreased nosZ abundance at 63% MC explained the observation of increased N2O concentration at that condition.

Accurate spring constant calibration for very stiff atomic force microscopy cantilevers.

There are many atomic force microscopy (AFM) applications that rely on quantifying the force between the AFM cantilever tip and the sample. The AFM does not explicitly measure force, however, so in such cases knowledge of the cantilever stiffness is required. In most cases, the forces of interest are very small, thus compliant cantilevers are used. A number of methods have been developed that are well suited to measuring low stiffness values. However, in some cases a cantilever with much greater stiffness is required. Thus, a direct, traceable method for calibrating very stiff (approximately 200 N/m) cantilevers is presented here. The method uses an instrumented and calibrated nanoindenter to determine the stiffness of a reference cantilever. This reference cantilever is then used to measure the stiffness of a number of AFM test cantilevers. This method is shown to have much smaller uncertainty than previously proposed methods. An example application to fracture testing of nanoscale silicon beam specimens is included.