Alan M. Lefcourt

Circadian and ultradian rhythms of body temperature and peripheral concentrations of insulin and nitrogen in lactating dairy cows

To investigate possible circadian and ultradian periodicities for peripheral insulin and urea in lactating dairy cows, integrated 15-min blood samples were taken sequentially over 48 hr from six cows. In addition, radiotelemetry measurements of body temperature were averaged over the same 15-min periods. Cows were housed in an environmental chamber at 19 degrees C with lights on 0700 to 2300 hr; fed daily at 0900 hr; and milked at 0800 and 2000 hr. For five of the six cows, body temperature showed a circadian rhythm peaking at 2323 hr with an amplitude of 0.34 degree C. For the sixth cow, body temperature was 180 degrees out-of-phase, peaking at 1230 hr with an amplitude of 0.12 degree C. Circadian rhythms for insulin and urea were consistent for all six cows peaking at 1743 hr with an amplitude of 0.74 ng/ml for insulin and at 1034 hr with an amplitude of 3.83 mM for urea. Body temperature and insulin also displayed episodic increases that often exceeded the amplitudes of circadian rhythms. For body temperature, a broad increase in spectral power was seen for periods between 100 and 175 min; time intervals between peaks averaged around 100 min. For insulin, power spectra for individual cows universally indicated rhythms with periods of approximately 45 and 80 min; time intervals between peaks averaged approximately 65 min. For urea, almost all spectral energy was confined to the 24-hr rhythm, although there was evidence of a low-amplitude, 60-min rhythm. In conclusion, when animals are acclimated to a rigidly controlled environment and frequent blood sampling is accomplished with minimal intervention, it is possible to detect rhythms inherent in the regulation of metabolic variables.

Multispectral laser-induced fluorescence imaging system for large biological samples

A laser-induced fluorescence imaging system developed to capture multispectral fluorescence emission images simultaneously from a relatively large target object is described. With an expanded, 355-nm Nd:YAG laser as the excitation source, the system captures fluorescence emission images in the blue, green, red, and far-red regions of the spectrum centered at 450, 550, 678, and 730 nm, respectively, from a 30-cm-diameter target area in ambient light. Images of apples and of pork meat artificially contaminated with diluted animal feces have demonstrated the versatility of fluorescence imaging techniques for potential applications in food safety inspection. Regions of contamination, including sites that were not readily visible to the human eye, could easily be identified from the images.

Milking- and suckling-induced secretion of oxytocin and prolactin in parturient dairy cows

Abstract Serum concentrations of prolactin were unaffected by either suckling or milking on Day 2 or 3 postpartum in cows housed with their calves following parturition. In contrast, among cows housed without their calves milking elicited a four- to sixfold increase in serum prolactin concentrations. Serum oxytocin levels increased in response to both suckling and milking among cows housed with their calves with suckling being a more potent stimulus (257 ± 32 vs 189 ± 23 pg/ml at peak). However, the greatest increase in oxytocin levels accompanied milking in cows housed without their calves (375 ± 36 pg/ml at peak). These results suggest that stimuli associated with the presence or the absence of the calf can alter maternal secretion of oxytocin and prolactin. Greater understanding of factors which regulate secretion of these hormones may result in techniques to modify milk synthesis and milk ejection in dairy cows.

Failure of cortisol injected prior to milking to inhibit milk ejection in dairy cattle

To test the potential for cortisol to inhibit milk ejection directly, 18 Holstein cows were divided equally into control and treatment groups based on milk yields. For treated animals, a single injection of cortisol was made into the saphenous vein 15 min before milkings. Increasing amounts of cortisol (0, 25, 50, and 100 mg) were injected for one morning and one evening milking, with the exception that the treated cows received only one 100 mg injection. Control animals received injections of 0.9% (w/v) NaCl. Cortisol injections had no effect on milk yields. However, a potential inhibitory mechanism might involve a delay, perhaps due to the necessity of synthesizing a regulatory protein. Therefore, to test the potential for increased cortisol over a period of hours to inhibit milk ejection, six of the nine cows in the treatment group were injected with 100 mg of cortisol at 3.25, 2.25, 1.25 and 0.25 h before sequential morning and evening milkings. In blood samples taken 1 min before and after injections, base-line cortisol concentrations averaged 10.2 mg/ml; after injection they were 984.1 ng/ml, and before subsequent injections they were 37.6 ng/ml. Again cortisol injections had no effect on milk yields.

Detection of fecal contamination on apples with nanosecond-scale time-resolved imaging of laser-induced fluorescence

Detection of apples contaminated with feces is a public health concern. We found that time-resolved imaging of apples artificially contaminated with feces allowed optimization of timing parameters for detection. Dairy feces were applied to Red Delicious and Golden Delicious apples. Laser-induced fluorescence responses were imaged by use of a gated intensified camera. We developed algorithms to automatically detect contamination iteratively by using one half of the apples and validated them by applying the optimized algorithms to the remaining apples. Results show that consideration of the timing of fluorescence responses to pulsed-laser excitation can enhance detection of feces on apples.

Multispectral fluorescence lifetime imaging of feces-contaminated apples by time-resolved laser-induced fluorescence imaging system with tunable excitation wavelengths

We recently developed a time-resolved multispectral laser-induced fluorescence (LIF) imaging system capable of tunable wavelengths in the visible region for sample excitation and nanosecond-scale characterizations of fluorescence responses (lifetime imaging). Time-dependent fluorescence decay characteristics and fluorescence lifetime imaging of apples artificially contaminated with a range of diluted cow feces were investigated at 670 and 685 nm emission bands obtained by 418, 530, and 630 nm excitations. The results demonstrated that a 670 nm emission with a 418 nm excitation provided the greatest difference in time-dependent fluorescence responses between the apples and feces-treated spots. The versatilities of the time-resolved LIF imaging system, including fluorescence lifetime imaging of a relatively large biological object in a multispectral excitation-emission wavelength domain, were demonstrated.

Safety Inspection of Cantaloupes and Strawberries Using Multispectral Fluorescence Imaging Techniques

Fluorescence is widely used for investigation of biological materials, and in recent years it has also been used to monitor food quality and safety. In this study the hyperspectral imaging system developed at the Instrumentation and Sensing Laboratory (ISL), United States Department of Agriculture (USDA) was used to evaluate the potential for detection of animal fecal contamination on cantaloupes and strawberries. Samples obtained from a local store were rinsed and allowed to air dry. Subsequently, fresh cow feces collected from the USDA dairy were diluted 1:10, 1:50, and 1:100 by weight with H2O and applied to the samples. Samples were excited using a UVA source and fluorescence responses were measured from 430 to 770 nm. Results indicate that fluorescence images at 680nm exhibited the greatest contrast between treated and untreated surfaces. For cantaloupes, contrast was improved using ratio images where images at 660 nm were divided by images at 560 nm. For strawberries, contrast was improved using the ratio of images at 680 nm and 745 nm. When data were analyzed for principle components (PC), the first four PC images exhibited useful results for contamination detection. Leaves present a problem when trying to identify fecal contamination on strawberries, however PC-3 provided contrast between them, creating ideal conditions for leaf masking. For cantaloupes, the PC-2 image differentiated contaminated regions. PC-1 presented distinctive differences between the contaminated spots and cantaloupe scarred tissue, suggesting possible use for such discrimination.

A Simple Multispectral Imaging Algorithm for Detection of Defects on Red Delicious Apples

Purpose: A multispectral algorithm for detection and differentiation of defective (defects on apple skin) and normal Red Delicious apples was developed from analysis of a series of hyperspectral line-scan images. Methods: A fast line-scan hyperspectral imaging system mounted on a conventional apple sorting machine was used to capture hyperspectral images of apples moving approximately 4 apples per second on a conveyor belt. The detection algorithm included an apple segmentation method and a threshold function, and was developed using three wavebands at 676 nm, 714 nm and 779 nm. The algorithm was executed on line-by-line image analysis, simulating online real-time line-scan imaging inspection during fruit processing. Results: The rapid multispectral algorithm detected over 95% of defectiv e apples and 91% of normal apples investigated. Conclusions: The multispectral defect detection algorithm can potentially be used in commercial apple processing lines.

A survey of unmanned ground vehicles with applications to agricultural and environmental sensing

Unmanned ground vehicles have been utilized in the last few decades in an effort to increase the efficiency of agriculture, in particular, by reducing labor needs. Unmanned vehicles have been used for a variety of purposes including: soil sampling, irrigation management, precision spraying, mechanical weeding, and crop harvesting. In this paper, unmanned ground vehicles, implemented by researchers or commercial operations, are characterized through a comparison to other vehicles used in agriculture, namely airplanes and UAVs. An overview of different trade-offs of configurations, control schemes, and data collection technologies is provided. Emphasis is given to the use of unmanned ground vehicles in food crops, and includes a discussion of environmental impacts and economics. Factors considered regarding the future trends and potential issues of unmanned ground vehicles include development, management and performance. Also included is a strategy to demonstrate to farmers the safety and profitability of implementing the technology.

Multispectral fluorescence imaging techniques for nondestructive food safety inspection

The use of spectral sensing has gained acceptance as a rapid means for nondestructive inspection of postharvest food produce. Current technologies generally use color or a single wavelength camera technology. The applicability and sensitivity of these techniques can be expanded through the use of multiple wavelengths. Reflectance in the Vis/NIR is the prevalent spectral technique. Fluorescence, compared to reflectance, is regarded as a more sensitive technique due to its dynamic responses to subtle changes in biological entities. Our laboratory has been exploring fluorescence as a potential means for detection of quality and wholesomeness of food products. Applications of fluorescence sensing require an understanding of the spectral characteristics emanating from constituents and potential contaminants. A number of factors affecting fluorescence emission characteristics are discussed. Because of relatively low fluorescence quantum yield from biological samples, a system with a powerful pulse light source such as a laser coupled with a gated detection device is used to harvest fluorescence, in the presence of ambient light. Several fluorescence sensor platforms developed in our laboratory, including hyperspectral imaging, and laser-induced fluorescence (LIF) and steady-state fluorescence imaging systems with multispectral capabilities are presented. We demonstrate the potential uses of recently developed fluorescence imaging platforms in food safety inspection of apples contaminated with animal feces.