Sub-10-nm-thick SiN nanopore membranes fabricated using the SiO2sacrificial layer process.

In our previous studies, ultrathin SiN membranes down to 3 nm in thickness were fabricated using the poly-Si sacrificial layer process, and nanopores were formed in those membranes. The region of the SiN membrane fabricated using this process was small, and the poly-Si sacrificial layer remained throughout the other region. On the other hand, to reduce the noise of the current through the nanopore, it is preferable to reduce the capacitance of the nanopore chip by replacing the poly-Si layer with an insulator with low permittivity, such as SiO2. Thus, in this study, the fabrication of SiN membranes with thicknesses of 3-7 nm using the SiO2sacrificial layer process was examined. SiN membranes with thicknesses of less than 5 nm could not be formed when the thickness of the top SiN layer deposited onto the sacrificial layer was 100 nm. In contrast, SiN membranes down to 3.07 nm in thickness could be formed when the top SiN layer was 40 nm in thickness. This is thought to be due to the difference in membrane stress. Nanopores were then fabricated in the membranes via dielectric breakdown. The current noise of the nanopore membranes was approximately 3/5 that of membranes fabricated using the poly-Si sacrificial layer process. Last, ionic current blockades were measured when poly(dT)60passed through the nanopores, and the effective thickness of the nanopores was estimated based on those current-blockade values. The effective thickness was approximately 4.8 nm when the deposited thickness of the SiN membrane was 6.03 nm. On the other hand, the effective thickness and the deposited thickness were almost the same when the deposited thickness was 3.07 nm. This suggests it became difficult to form a shape in which the thickness of the nanopore edge was thinner than the deposited membrane thickness as the deposited thickness decreased.

Artificial Intelligence-assisted System Improves Endoscopic Identification of Colorectal Neoplasms.

BACKGROUND & AIMS: Precise optical diagnosis of colorectal polyps could improve the cost-effectiveness of colonoscopy and reduce polypectomy-related complications. However, it is difficult for community-based non-experts to obtain sufficient diagnostic performance. Artificial intelligence-based systems have been developed to analyze endoscopic images; they identify neoplasms with high accuracy and low interobserver variation. We performed a multi-center study to determine the diagnostic accuracy of EndoBRAIN, an artificial intelligence-based system that analyzes cell nuclei, crypt structure, and microvessels in endoscopic images, in identification of colon neoplasms. METHODS: The EndoBRAIN system was initially trained using 69,142 endocytoscopic images, taken at 520-fold magnification, from patients with colorectal polyps who underwent endoscopy at 5 academic centers in Japan from October 2017 through March 2018. We performed a retrospective comparative analysis of the diagnostic performance of EndoBRAIN vs that of 30 endoscopists (20 trainees and 10 experts); the endoscopists assessed images from 100 cases produced via white-light microscopy, endocytoscopy with methylene blue staining, and endocytoscopy with narrow-band imaging. EndoBRAIN was used to assess endocytoscopic, but not white-light, images. The primary outcome was the accuracy of EndoBrain in distinguishing neoplasms from non-neoplasms, compared with that of endoscopists, using findings from pathology analysis as the reference standard. RESULTS: In analysis of stained endocytoscopic images, EndoBRAIN identified colon lesions with 96.9% sensitivity (95% CI, 95.8%-97.8%), 100% specificity (95% CI, 99.6%-100%), 98% accuracy (95% CI, 97.3%-98.6%), a 100% positive-predictive value (95% CI, 99.8%-100%), and a 94.6% negative-predictive (95% CI, 92.7%-96.1%); these values were all significantly greater than those of the endoscopy trainees and experts. In analysis of narrow-band images, EndoBRAIN distinguished neoplastic from non-neoplastic lesions with 96.9% sensitivity (95% CI, 95.8-97.8), 94.3% specificity (95% CI, 92.3-95.9), 96.0% accuracy (95% CI, 95.1-96.8), a 96.9% positive-predictive value, (95% CI, 95.8-97.8), and a 94.3% negative-predictive value (95% CI, 92.3-95.9); these values were all significantly higher than those of the endoscopy trainees, sensitivity and negative-predictive value were significantly higher but the other values are comparable to those of the experts. CONCLUSIONS: EndoBRAIN accurately differentiated neoplastic from non-neoplastic lesions in stained endocytoscopic images and endocytoscopic narrow-band images, when pathology findings were used as the standard. This technology has been authorized for clinical use by the Japanese regulatory agency and should be used in endoscopic evaluation of small polyps more widespread clinical settings. UMIN clinical trial no: UMIN000028843.

Solid-state nanopores towards single-molecule DNA sequencing.

Nanopore DNA sequencing offers a new paradigm owing to its extensive potential for long-read, high-throughput detection of nucleotide modification and direct RNA sequencing. Given the remarkable advances in protein nanopore sequencing technology, there is still a strong enthusiasm in exploring alternative nanopore-sequencing techniques, particularly those based on a solid-state nanopore using a semiconductor material. Since solid-state nanopores provide superior material robustness and large-scale integrability with on-chip electronics, they have the potential to surpass the limitations of their biological counterparts. However, there are key technical challenges to be addressed: the creation of an ultrasmall nanopore, fabrication of an ultrathin membrane, control of the ultrafast DNA speed and detection of four nucleotides. Extensive research efforts have been devoted to resolving these issues over the past two decades. In this review, we briefly introduce recent updates regarding solid-state nanopore technologies towards DNA sequencing. It can be envisioned that emerging technologies will offer a brand new future in DNA-sequencing technology.

Endocytoscopy for the differential diagnosis of colorectal low-grade adenoma: a novel possibility for the "resect and discard" strategy.

BACKGROUND AND AIMS: Endocytoscopy, a next-generation endoscopic system, facilitates observation at a maximum magnification of ×520. To our knowledge, no study has reported high-precision diagnosis of colorectal low-grade adenoma, endoscopically. We aimed to reveal which endocytoscopic findings may be used as indicators of low-grade adenoma and to assess whether a "resect and discard" strategy using endocytoscopy is feasible. METHODS: Lesions diagnosable with endocytoscopy were examined retrospectively between May 2005 and July 2017. A normal pit-like structure in endocytoscopic images was considered a normal pit (NP) sign and used as an indicator of low-grade adenoma. The primary outcome was the diagnostic accuracy of the NP sign for low-grade adenoma. We evaluated agreement rates between endocytoscopic and pathologic diagnosis for surveillance colonoscopy interval recommendation (SCIR) and performed a validation study to verify the agreement rates. RESULTS: For 748 lesions in 573 cases diagnosed as colorectal adenoma using endocytoscopy, the results were as follows: sensitivity of the NP sign for low-grade adenoma, 85.0%; specificity, 90.7%; positive predictive value, 96.6%; negative predictive value, 66.1%; accuracy, 86.4%; and positive likelihood ratio, 9.2 (P < .001). The agreement rate between endocytoscopic and pathologic diagnosis for SCIR was 94.4% (95% confidence interval [CI], 92.2%-96.1%; P < .001) under United States guidelines and 96.3% (95% CI, 94.5%-97.7%; P < .001) under European Union guidelines. All inter- and intraobserver agreement rates for expert and nonexpert endoscopists had κ values ≥0.8 except one nonexpert pair. CONCLUSIONS: Endocytoscopy is an effective modality in determining the differential diagnosis of colorectal low-grade adenoma. (University Hospital Medical Information Network Clinical Trials database registration number: UMIN000018623.).

Real-Time Use of Artificial Intelligence in Identification of Diminutive Polyps During Colonoscopy: A Prospective Study.

Background: Computer-aided diagnosis (CAD) for colonoscopy may help endoscopists distinguish neoplastic polyps (adenomas) requiring resection from nonneoplastic polyps not requiring resection, potentially reducing cost. Objective: To evaluate the performance of real-time CAD with endocytoscopes (×520 ultramagnifying colonoscopes providing microvascular and cellular visualization of colorectal polyps after application of the narrow-band imaging [NBI] and methylene blue staining modes, respectively). Design: Single-group, open-label, prospective study. (UMIN [University hospital Medical Information Network] Clinical Trial Registry: UMIN000027360). Setting: University hospital. Participants: 791 consecutive patients undergoing colonoscopy and 23 endoscopists. Intervention: Real-time use of CAD during colonoscopy. Measurements: CAD-predicted pathology (neoplastic or nonneoplastic) of detected diminutive polyps (≤5 mm) on the basis of real-time outputs compared with pathologic diagnosis of the resected specimen (gold standard). The primary end point was whether CAD with the stained mode produced a negative predictive value (NPV) of 90% or greater for identifying diminutive rectosigmoid adenomas, the threshold required to "diagnose-and-leave" nonneoplastic polyps. Best- and worst-case scenarios assumed that polyps lacking either CAD diagnosis or pathology were true- or false-positive or true- or false-negative, respectively. Results: Overall, 466 diminutive (including 250 rectosigmoid) polyps from 325 patients were assessed by CAD, with a pathologic prediction rate of 98.1% (457 of 466). The NPVs of CAD for diminutive rectosigmoid adenomas were 96.4% (95% CI, 91.8% to 98.8%) (best-case scenario) and 93.7% (CI, 88.3% to 97.1%) (worst-case scenario) with stained mode and 96.5% (CI, 92.1% to 98.9%) (best-case scenario) and 95.2% (CI, 90.3% to 98.0%) (worst-case scenario) with NBI. Limitation: Two thirds of the colonoscopies were conducted by experts who had each experienced more than 200 endocytoscopies; 186 polyps not assessed by CAD were excluded. Conclusion: Real-time CAD can achieve the performance level required for a diagnose-and-leave strategy for diminutive, nonneoplastic rectosigmoid polyps. Primary Funding Source: Japan Society for the Promotion of Science.

Artificial intelligence and colonoscopy: Current status and future perspectives.

BACKGROUND AND AIM: Application of artificial intelligence in medicine is now attracting substantial attention. In the field of gastrointestinal endoscopy, computer-aided diagnosis (CAD) for colonoscopy is the most investigated area, although it is still in the preclinical phase. Because colonoscopy is carried out by humans, it is inherently an imperfect procedure. CAD assistance is expected to improve its quality regarding automated polyp detection and characterization (i.e. predicting the polyp's pathology). It could help prevent endoscopists from missing polyps as well as provide a precise optical diagnosis for those detected. Ultimately, these functions that CAD provides could produce a higher adenoma detection rate and reduce the cost of polypectomy for hyperplastic polyps. METHODS AND RESULTS: Currently, research on automated polyp detection has been limited to experimental assessments using an algorithm based on ex vivo videos or static images. Performance for clinical use was reported to have >90% sensitivity with acceptable specificity. In contrast, research on automated polyp characterization seems to surpass that for polyp detection. Prospective studies of in vivo use of artificial intelligence technologies have been reported by several groups, some of which showed a >90% negative predictive value for differentiating diminutive (≤5 mm) rectosigmoid adenomas, which exceeded the threshold for optical biopsy. CONCLUSION: We introduce the potential of using CAD for colonoscopy and describe the most recent conditions for regulatory approval for artificial intelligence-assisted medical devices.

Artificial intelligence may help in predicting the need for additional surgery after endoscopic resection of T1 colorectal cancer.

BACKGROUND AND STUDY AIMS: Decisions concerning additional surgery after endoscopic resection of T1 colorectal cancer (CRC) are difficult because preoperative prediction of lymph node metastasis (LNM) is problematic. We investigated whether artificial intelligence can predict LNM presence, thus minimizing the need for additional surgery. PATIENTS AND METHODS: Data on 690 consecutive patients with T1 CRCs that were surgically resected in 2001 - 2016 were retrospectively analyzed. We divided patients into two groups according to date: data from 590 patients were used for machine learning for the artificial intelligence model, and the remaining 100 patients were included for model validation. The artificial intelligence model analyzed 45 clinicopathological factors and then predicted positivity or negativity for LNM. Operative specimens were used as the gold standard for the presence of LNM. The artificial intelligence model was validated by calculating the sensitivity, specificity, and accuracy for predicting LNM, and comparing these data with those of the American, European, and Japanese guidelines. RESULTS: Sensitivity was 100 % (95 % confidence interval [CI] 72 % to 100 %) in all models. Specificity of the artificial intelligence model and the American, European, and Japanese guidelines was 66 % (95 %CI 56 % to 76 %), 44 % (95 %CI 34 % to 55 %), 0 % (95 %CI 0 % to 3 %), and 0 % (95 %CI 0 % to 3 %), respectively; and accuracy was 69 % (95 %CI 59 % to 78 %), 49 % (95 %CI 39 % to 59 %), 9 % (95 %CI 4 % to 16 %), and 9 % (95 %CI 4 % - 16 %), respectively. The rates of unnecessary additional surgery attributable to misdiagnosing LNM-negative patients as having LNM were: 77 % (95 %CI 62 % to 89 %) for the artificial intelligence model, and 85 % (95 %CI 73 % to 93 %; P < 0.001), 91 % (95 %CI 84 % to 96 %; P < 0.001), and 91 % (95 %CI 84 % to 96 %; P < 0.001) for the American, European, and Japanese guidelines, respectively. CONCLUSIONS: Compared with current guidelines, artificial intelligence significantly reduced unnecessary additional surgery after endoscopic resection of T1 CRC without missing LNM positivity.

Accuracy of diagnosing invasive colorectal cancer using computer-aided endocytoscopy.

Background and study aims Invasive cancer carries the risk of metastasis, and therefore, the ability to distinguish between invasive cancerous lesions and less-aggressive lesions is important. We evaluated a computer-aided diagnosis system that uses ultra-high (approximately × 400) magnification endocytoscopy (EC-CAD). Patients and methods We generated an image database from a consecutive series of 5843 endocytoscopy images of 375 lesions. For construction of a diagnostic algorithm, 5543 endocytoscopy images from 238 lesions were randomly extracted from the database for machine learning. We applied the obtained algorithm to 200 endocytoscopy images and calculated test characteristics for the diagnosis of invasive cancer. We defined a high-confidence diagnosis as having a ≥ 90 % probability of being correct. Results Of the 200 test images, 188 (94.0 %) were assessable with the EC-CAD system. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) were 89.4 %, 98.9 %, 94.1 %, 98.8 %, and 90.1 %, respectively. High-confidence diagnosis had a sensitivity, specificity, accuracy, PPV, and NPV of 98.1 %, 100 %, 99.3 %, 100 %, and 98.8 %, respectively. Conclusion: EC-CAD may be a useful tool in diagnosing invasive colorectal cancer.

Double staining with crystal violet and methylene blue is appropriate for colonic endocytoscopy: an in vivo prospective pilot study.

BACKGROUND AND AIM: Endocytoscopy (EC) at ultra-high magnification enables in vivo visualization of cellular atypia of gastrointestinal mucosae. Clear images are essential for precise diagnosis by EC. The aim of the present study was to evaluate the optimal staining method for EC in the colon. METHODS: Thirty prospectively enrolled patients were allocated 1:1:1 to three distinct staining methods: 0.05% crystal violet (CV) alone, 1% methylene blue (MB) alone, or CV+MB (CM double). Normal rectal mucosae were stained with each dye and videos of EC images were recorded. Visibility of nuclei and gland formation after staining were evaluated as 'recognizable' or 'not recognizable'. Time for each parameter to become 'recognizable' was measured, and the average times for the three staining regimens were compared. RESULTS: MB alone and CM double staining resulted in 'recognizable' (102 ± 27 vs 89 ± 22 s, P=0.263) nuclei within comparable periods of time, whereas CV alone was unable to identify nuclei. Gland formation became 'recognizable' sooner after CM double staining than after MB alone (61 ± 16 vs 108 ± 24 s, P<0.001). CONCLUSIONS: Double staining with CV and MB, which rapidly provided recognizable images of both nuclei and gland formation, is an appropriate staining regimen for colonic EC.

Effects of one-hour daily outdoor access on milk yield and composition and behaviors of tethered dairy cows.

Objective: We investigated the effects of outdoor access for one-hour per day (ODA) on milk yield and composition and behaviors of tethered dairy cows. Methods: Eleven all-day tethered dairy cows were treated the ODA for two weeks. To evaluate the effect of ODA on milk yield, we first calculated the average daily milk yield of each cow for three days during two weeks before the ODA, three days before the ODA, three days at the end of the ODA, and three days during two weeks after the ODA. We then compared the milk yield change during the ODA with that for two weeks before and two weeks after the ODA. The effects of ODA on milk compositions and behaviors were evaluated by comparing the average values for each composition and behavior for the three days before the ODA and the last three days of the ODA. Results: The decrease of milk yield during the two weeks of ODA was significantly higher than that during the two weeks before ODA (p<0.01). The milk fat rate was significantly higher during ODA than before ODA (p<0.05). Lactose rate was significantly lower during ODA than before ODA (p<0.05). The concentrations of milk urea nitrogen, ketone bodies, and free fatty acids in the milk were significantly higher during ODA than before ODA (p<0.05). The mean total duration per day of lying during ODA was significantly lower than that before ODA (p<0.05). The walking steps per one-hour outdoor access were 158.4 ± 54.7. The social behavior during the one-hour outdoor access of the 11 cows was 53 times/h/herd. . Conclusion: Our results suggested that ODA promotes the express of normal behavior in dairy cows, but even one hour of ODA decreases milk production in cows, which may drop producers' profits without some financial supplementation.

Dwell Time Prolongation and Identification of Single Nucleotides Passing through a Solid-State Nanopore by Using Ammonium Sulfate Aqueous Solution.

The ionic current blockades when poly(dT)60 or dNTPs passed through SiN nanopores in an aqueous solution containing (NH4)2SO4 were investigated. The dwell time of poly(dT)60 in the nanopores in an aqueous solution containing (NH4)2SO4 was significantly longer compared to that in an aqueous solution that did not contain (NH4)2SO4. This dwell time prolongation effect due to the aqueous solution containing (NH4)2SO4 was also confirmed when dCTP passed through the nanopores. In addition, when the nanopores were fabricated via dielectric breakdown in the aqueous solution containing (NH4)2SO4, the dwell time prolongation effect for dCTP still occurred even after the aqueous solution was displaced with the aqueous solution without (NH4)2SO4. Furthermore, we measured the ionic current blockades when the four types of dNTPs passed through the same nanopore, and the four types of dNTPs could be statistically identified according to their current blockade values.

Effects of one hour daily outdoor access on lying and sleeping postures, and immune traits of tethered cows.

OBJECTIVE: We investigated the effects of outdoor access for 1 h per day on the animal welfare (AW) of tethered cows, in terms of lying and sleeping postures, and immune function. METHODS: A total of five dry cows were tethered all day indoors (tethering) for 30 days and then tethered indoors with 1 h daily outdoor access (ODA-1h) for 30 days. To analyze the effects of ODA-1h, we calculated the total duration and bout frequency per day, and bout duration of lying and sleeping postures during the last five days of each treatment period. We also analyzed the populations of T cells, B cells, and NK cells in peripheral blood mononuclear cells (PBMC) by fluorescence-activated cell sorting and determined the concanavalin A (Con A) -induced proliferation rate of T cells. RESULTS: The mean total time per day of lying during the ODA-1h treatment was significantly shorter than that during the tethering treatment (p<0.001). The Con A-induced proliferation rate of T cells during the ODA-1h treatment was significantly higher than that during the tethering treatment (p = 0.007). The proportion of NK cells in PBMC during the ODA-1h treatment tended to be higher than that during the tethering treatment (p = 0.062). CONCLUSION: Although ODA-1h may decrease lying time, it increases the available space for tethered cows towards that typically found in grazing and free barn feeding systems. This increased available space promotes the expression of normal behaviors such as walking and social behaviors except lying and may also improve the immune function of tethered dry cows, thereby improving their overall welfare.

Meat the challenge: Segmentation and profiling of Japanese beef mince and its substitutes consumers.

Shifts in protein production methods are an emerging challenge toward realizing a sustainable society. This paper aims to examine preferences among Japanese consumers regarding attributes of beef mince and its substitutes, to develop consumer segments based on these preferences, and to explore the segment with higher acceptance of replacement from conventional products. This paper also aims to explain intersegment differences from consumer heterogeneity in human values, scientific literacy, and sociodemographic viewpoints for a deeper understanding of consumer behavior in each segment. The results of an online choice experiment involving 4421 consumers in Japan, using food labels on mince showed that Japanese-origin organic beef was associated with the highest utility among the five production methods mentioned. Five consumer segments were identified with latent class analysis: novelty accepters, generous customers, attribute-economy balancers, price-conscious, and conservatives, which vary in preference in choice behavior, sociodemographic, human values, and scientific literacy.

Effect of different cow floor mat on the pressure in anterior knees of Holstein dry cows determined using pressure measurement sheet.

To analyze limb pain in tethered dairy cows, we compared three cow floor mat materials (pasture mat® [PM], KKM rubber mat® [RM], and woodchip [WC]) with soil ground considering the pressure applied to the anterior knee of Holstein cows. Three Holstein dry cows (791.3 ± 44.6 kg, 3.8 ± 1.1 years old) moored at the Education and Research Center of Alpine Field Science of the Faculty of Agriculture, Shinshu University, were tested. Pressure applied to the anterior knee of the cow by four different floor mat materials was compared using a commercially available pressure measurement sheet. The sustained pressures applied to the anterior knee of the test cow were 7.2 ± 1. 5, 29.3 ± 5.1, 17.0 ± 1.4, and 27.7 ± 2.5 MPa in the control, RM, PM, and WC, respectively, showing significant lower values (P < 0.05) than in the control. Sustained pressure was significantly lower in the PM than in the other treatments (P < 0.05). The RM was 41.7% cheaper than the PM. The pressure applied to the anterior knee of cow is lower on PM than on RM and WC. PM was more costly than RM but the sustained pressure was much lower on PM.

Effects of starvation-induced negative energy balance on endoplasmic reticulum stress in the liver of cows.

OBJECTIVE: Endoplasmic reticulum (ER) stress engages the unfolded protein response (UPR) that serves as an important mechanism for modulating hepatic fatty acid oxidation and lipogenesis. Chronic fasting in mice induced the UPR activation to regulate lipid metabolism. However, there is no direct evidence of whether negative energy balance (NEB) induces ER stress in the liver of cows. This study aimed to elucidate the relationship between the NEB attributed to feed deprivation and ER stress in bovine hepatocytes. METHODS: Blood samples and liver biopsy tissues were collected from 6 non-lactating cows before and after their starvation for 48 h. The blood non-esterified fatty acids (NEFA), ß-hydroxybutyric acid (BHBA) and glucose level were analyzed. Real-time quantitative polymerase chain reaction and Western blotting were used to explore the regulation of genes associated with UPR and lipid metabolism. RESULTS: The starvation increased the plasma BHBA and NEFA levels and decreased the glucose level. Additionally, the starvation caused significant increases in the mRNA expression level of spliced X-box binding protein 1 (XBP1s) and the protein level of phosphorylated inositol-requiring kinase 1 alpha (p-IRE1α; an upstream protein of XBP1) in the liver. The mRNA expression levels of peroxisome proliferator-activated receptor alpha and its target fatty acid oxidation- and ketogenesis-related genes were significantly upregulated by the starvation-mediated NEB. Furthermore, we found that the mRNA expression levels of lipogenic genes were not significantly changed after starvation. CONCLUSION: These findings suggest that in the initial stage of NEB in dairy cows, the liver coordinates an adaptive response by activating the IRE1 arm of the UPR to enhance ketogenesis, thereby avoiding a fatty liver status.

[A case of organizing pneumonia with a reversed halo sign following postoperative irradiation for breast cancer].

A 69-year-old woman received radiation therapy of 50 Gy following surgery for left breast cancer. Eleven months later, chest computed tomography revealed infiltrative shadows with air bronchograms in both lower lung fields and a reversed halo sign in the right lower lobe. Transbronchial lung biopsy revealed polypoid granulation tissue in the terminal air spaces, which was consistent with organizing pneumonia (OP). Prednisolone therapy resolved the radiographic abnormalities. The reversed halo sign was believed to be specific to cryptogenic organizing pneumonia. We reported a case of OP with a reversed halo sign following postoperative irradiation for breast cancer.

Effects of adding a commercial capsaicin fertilizer on the feeding behavior of sika deer (Cervus nippon).

We investigated the inhibitory effect of capsaicin fertilizer on feeding in deer. We tested four captive adult female deer. In Experiment 1, in addition to the treatment (intact) containing only a solid feed (HC), we mixed the fertilizer not containing capsaicin (F) or the capsaicin fertilizer (CF) in the solid feed. In addition, the solid feed was put on a wire net that capsaicin fertilizer was placed 5 cm below (SCF). We investigated their feeding behavior response. In Experiment 2, we changed the amount of substance (fertilizer and capsaicin fertilizer) mixed in the HC. We mixed different amounts (0, 50, 100, and 200 g) of the treatments other than the intact with HC and presented them to the deer, and investigated their feeding behavior response. In Experiment 1, intake in the F and CF decreased (p < .05). In Experiment 2, HC intake was significantly lower in the 100 and 200 g CF (p < .05). However, HC intake relatively increased by the last day in the CF 200 g too. The capsaicin fertilizer decreased the feeding behavior of deer by directly touching the mucous membranes of the deer nose and lips. However, the effects were decreased over time.