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The objectives of this study were to evaluate the results when tissue-engineered vascular grafts (TEVGs) are used as alternatives to autologous pericardium for surgically augmenting the pulmonary artery (PA) or aortic valve.TEVG molds were embedded into subcutaneous spaces for more than 4 weeks preoperatively. Since 2014, 6 patients have undergone PA reconstruction, whereas 1 has undergone aortic valve plasty (AVP) with TEVGs. The time from mold implantation to the operation was 8.9 (range, 6.0-26.4) months. The age and body weight at the time of operation were 2.7 (range, 1.8-9.2) and 11.6 (range, 7.9-24.4) kg, respectively. Concomitant procedures comprised the Rastelli, palliative Rastelli, and Fontan operations in 2, 2, and 1 patient, respectively.The median follow-up period was 14.4 (range, 3-39.6) months. There were no early or late mortalities. Moreover, there were no TEVG-related complications, including aneurysmal changes, degeneration, and infection. In 5 patients who underwent PA augmentation, the postoperative PA configuration was satisfactorily dilated. The reconstructed aortic valve function was good in the patient who underwent AVP. Decreased leaflet flexibility due to leaflet thickening was not observed. One patient had postoperative PA re-stenosis; therefore, re-PA augmentation with TEVGs was performed. On histological examination, TEVGs consisted of collagen fibers and few fibroblasts, and elastic fiber formation and/or smooth muscle cells were not observed.The midterm results of PA reconstruction and AVP with TEVGs were satisfactory. TEVGs might be a useful alternative to autologous pericardium in pediatric cardiovascular surgeries that often require multistage operations.

We developed a handmade expanded polytetrafluoroethylene (ePTFE) pulmonary valvular conduit (PVC) with bulging sinuses and a fan-shaped ePTFE valve for right ventricular outflow tract (RVOT) reconstruction. We aimed to investigate the results of this device in this multicenter study. From 2001 to 2020, 1776 patients underwent RVOT reconstruction using ePTFE PVCs at 65 institutions in Japan. The median age and body weight were 4.1 years (range, 3 days to 67.1 years) and 13.3 (range, 1.8-91.3) kg, respectively. The median PVC size was 18 (range, 8-24) mm. The median Z-value of the ePTFE PVC was 1.1 (range, -3.8 to 5.0). The ePTFE PVC conditions were investigated by cardiac echocardiography and catheterization. The median follow-up period was 3.3 years (range, 0 day to 16.2 years). There were only 9 cases (0.5%) with PVC-related unknown deaths. Reintervention was performed in 283 patients (15.9%), and 190 patients (10.7%) required explantation. Freedom from reintervention and explantation at 5/10 years were 86.7/61.5% and 93.0/69.1%, respectively. At the latest echocardiography, PVC regurgitation grade was better than mild in 88.4% patients. The average peak RVOT gradient was 15.7 ± 15.9 mm Hg at the latest cardiac catheterization. ePTFE PVC infection was detected in only 8 patients (0.5%). Relative stenosis due to somatic growth was the most common cause of PVC explantation. The performance of ePTFE in terms of durability, valvular performance, and the resistance against infection is considerable and may replace conventional prosthetic materials. Further improvement of the ePTFE membrane is essential to prevent valvular dysfunction.

Cone reconstruction for Ebstein’s anomaly, although effective, is challenging in neonatal cases. Very few reports have discussed recurrent regurgitation. Herein, we report a successful redo tricuspid valve repair for recurrent regurgitation 5 years after cone reconstruction. A 5-year-old boy underwent cone reconstruction for Ebstein’s anomaly in the neonatal period. Although tricuspid regurgitation reduced initially, it subsequently worsened. The mechanisms of regurgitation are dilatation of the anteroseptal commissure, indentation in the septal leaflet, and foreshortening of the anterior leaflet. Augmentation of the anterior leaflet using an elliptic autologous pericardium and mending of the gaping commissure and indentation markedly reduced the regurgitation.

OBJECTIVES Pulmonary artery (PA) compression by the neoaorta is a serious complication after the Norwood-type palliation (NP) for hypoplastic left heart syndrome. Either excess patch tailoring or limited use of autologous tissue may cause narrowing of the subaortic space. The chimney technique could theoretically provide a wide subaortic space. METHODS Twenty-nine patients with both pre- and post-NP computed tomography data available of the 37 consecutive patients who underwent NP in our institution were reviewed. Arch height, arch width, sinus of Valsalva diameter, area under the neoaortic arch and arch angle were measured. These patients were divided according to the neo-arch reconstruction technique, chimney reconstruction technique (CR) or conventional direct reconstruction technique (DR). RESULTS Median age and weight at NP were 2.1 months and 3.5 kg, respectively. Twenty-two patients underwent previous bilateral PA banding. During NP, 17 CR and 12 DR were performed. Four patients in the DR group developed PA compression. No neoaortic arch dilatation was found in either group. Post-NP arch width, area under the neo-arch and the arch angle were significantly larger in the CR group. Pre-NP arch height was significantly smaller in DR patients with PA compression than in those without. CONCLUSIONS The chimney technique decreased the risk of PA compression and provided a wider subaortic space and a less acute arch angle. This technique had no short-term effect on the neoaortic root. Small preoperative arch height is a potential risk factor for PA compression in DR, and the chimney technique could be an effective solution.

We evaluated the application of in vivo tissue-engineered vascular graft (in vivo TEVG) in pulmonary artery (PA) reconstruction as a substitute for autologous pericardium. From July 2017 to April 2020, 4 patients (male:female = 2:2) with major aortopulmonary collateral arteries underwent PA reconstruction with in vivo TEVGs. Graft moulds were embedded into the subcutaneous spaces in the first palliative surgery. In the second surgery used in vivo TEVGs were used as patch materials to treat PA stenosis. Preoperative and postoperative PA configurations were evaluated by computed tomography. Patients’ median age and body weight were 1.6 (1–4) years and 8.7 (7.3–15.4) kg, respectively. Two patients underwent PA reconstruction during staged repair and 2 underwent reconstruction during definitive repair. One patient had postoperative PA restenosis due to bronchial compression; re-PA reconstruction with in vivo TEVGs was performed. On histological examination, the in vivo TEVG wall mainly comprised collagen fibres and a small number of fibroblasts. The midterm results of this technique are satisfactory. in vivo TEVGs could be a promising alternative to autologous pericardium for paediatric cardiovascular surgeries requiring multi-stage operations. Clinical trial registration ERB-C-162

OBJECTIVES The aim of this study was to clarify the impact of valved systemic ventricle–pulmonary artery (SV–PA) shunt on outcomes after stage-1 Norwood-type palliation (NP) compared with the modified Blalock–Taussig shunt. METHODS Consecutive patients who underwent NP between 2003 and 2019 were enrolled. SV–PA shunts using the expanded polytetrafluoroethylene valved conduit were implanted in 18 patients (valved SV–PA group), and another 18 patients underwent modified Blalock–Taussig shunt during NP (modified Blalock–Taussig shunt group). All valved conduits were made in our institution in advance. RESULTS No differences in baseline characteristics were found between the groups, except for shunt size. During a median 2.9 (interquartile range 0.4–6.4, maximum 14.2) years of follow-up, 8 (22.2%) patients died across both groups. There were no statistically significant differences in early mortality (5.5% vs 11.1%, P = 0.55) and overall survival rates at 5 years (80.8% vs 71.4%, P = 0.48) in the valved SV–PA and modified Blalock–Taussig shunt groups. No statistically significant difference was observed in the frequency of interventions between the groups (31% vs 33%, P = 1.0). At the time of the bidirectional Glenn procedure, the systemic ventricular end-diastolic volume index was significantly lower (84 ± 24 vs 106 ± 31 ml/m2, P = 0.05) and the ejection fraction was significantly greater (62 ± 8% vs 55 ± 9%, P = 0.03) in the valved SV–PA group. There was no statistically significant difference in the pulmonary artery index (228 ± 85 vs 226 ± 60 mm2/m2, P = 0.92). CONCLUSIONS A valved SV–PA shunt using an expanded polytetrafluoroethylene valved conduit was associated with preserved ventricular function after NP and did not impair pulmonary artery growth by controlling pulmonary regurgitation.

The aim of study was to investigate the relationships among serum diamine oxidase (DAO) activity, postnatal days and the plasma copper (Cu) concentration, using calves with or without diarrhea. In healthy calves, the serum DAO activity was significantly higher at 2 postnatal days than at ≥7 postnatal days, and no significant changes were observed after 7 postnatal days. In addition, no significant correlation was found between serum DAO activity and plasma Cu concentration at all postnatal days in healthy calves. Although, the serum DAO activity in 14 diarrheic calves (66.78 ± 14.37 IU/ml) was lower than that in 19 healthy calves (170.33 ± 97.83 IU/ml, P

The aim of the present study was to investigate whether abnormalities in plasma diamine oxidase (DAO) activity reflect the degree of intestinal mucosal disorder in calves with diarrhea. A total of 50 Holstein calves were enrolled. Thirty-six of the 50 calves presented diarrhea and were sub-classified by severity based on fecal status (0: firm, 1: pasty, 2: loose, and 3: watery) and blood pH (acidemia: blood pH

The present study examined the effects of substituting kraft pulp (KP) with corn silage (CS) on dry matter intake (DMI), ruminal mat and rumen fermentation characteristics, and rumination. Four non-lactating, rumen-cannulated Holstein cows were fed a CS diet comprising 36% grass silage (GS) and 64% CS or a KP diet comprising 36% GS, 57% KP, and 7% soybean meal. DMI was significantly lower in cows fed the KP diet than in those fed the CS diet (p < 0.05), whereas rumination time did not significantly differ between the treatments. Dry matter content in the rumen immediately before and 3 h after feeding was significantly higher in cows fed the KP diet than in those fed the CS diet (p < 0.05). The consistency and thickness of the ruminal mat did not significantly differ between the treatments. The ruminal mean retention time of feed particles tended to be longer in cows fed the KP diet than in those fed the CS diet (p < 0.10). The ruminal digestion rate of KP was comparable to that of GS and CS. Because ruminal mat was formed and rumination was stimulated, KP was considered to have the equivalent physical effectiveness as CS.

OBJECTIVES To compare conventional procedures with the half-turned truncal switch operation (HTTSO) for the management of complete transposition of the great arteries with left ventricular outflow tract (LVOT) obstruction using time-resolved 3-dimensional magnetic resonance phase-contrast imaging. METHODS We identified 2 cases that underwent the Rastelli procedure and one case that underwent the Réparation a l'étage ventriculaire before 2002 [conventional procedures group (group C)], and 16 cases of HTTSO that were performed between 2002 and 2020 [HTTSO group (group H)]. Postoperative haemodynamics were assessed using time-resolved 3-dimensional magnetic resonance phase-contrast imaging in cases in both groups. RESULTS The median follow-up period was 20.4 years in group C, and 6.1 years in group H. In group C, all 3 patients underwent reoperation because of postoperative right ventricular outflow tract obstruction and/or insufficiency. In addition, permanent pacemaker implantation was needed in 1 patient because of complete atrioventricular block complicated by ventricular septal defect enlargement. In group H, reoperation for LVOT/right ventricular outflow tract obstruction was not needed. A time-resolved 3-dimensional magnetic resonance phase-contrast imaging examination revealed high energy loss and wall shear stress in the winding LVOT in the group C. In contrast, low energy loss and wall shear stress, with straight and smooth LVOT, were identified in group H. CONCLUSIONS HTTSO was shown to be superior to conventional procedures because a straight and wide LVOT could be obtained. Therefore, HTTSO should be the first choice for complete transposition of the great arteries with LVOT obstruction.

OBJECTIVES Late complications of arterial switch operations (ASO) for transposition of the great arteries, such as neo-pulmonary artery (PA) stenosis and/or neoaortic regurgitation, have been reported. We developed an alternative reconstruction method called the longitudinal extension (LE) method to prevent PA bifurcation stenosis (PABS). METHODS We identified 48 patients diagnosed with transposition of the great arteries and performed ASO using the Lecompte manoeuvre for neo-PA reconstruction. In 9 consecutive patients (from 2014), the LE method was performed (LE). Before 2014, conventional techniques were performed in 39 patients (C). The median body weight and age in the LE and C groups were 3.0 and 3.1 kg and 12 and 26 days, respectively. In the LE group, 1 patient underwent bilateral PA banding before ASO. In C, PA banding and arch repair were performed in 1 patient each. Patients who received concomitant procedures were included. RESULTS The median follow-up in LE and C groups was 1.9 and 10.1 years, respectively. Early mortality/late death was not found in group LE and in 1 patient in group C. Only 1 case required ascending aorta sliding plasty in LE, and 8 patients needed PA augmentation for PABS in C. The median velocity of right/left PA was measured as 1.6/1.9 m/s in LE and 2.1/2.3 m/s in C, so it showed a lower value in LE. CONCLUSIONS Excellent mid-term results were obtained with the LE method. It was considered a useful procedure in preventing PABS, which is a primary late complication of ASO. Further follow-up and investigations are needed.

OBJECTIVES In Japan, homograft and bovine jugular vein are available in very limited institutions for the reconstruction of the right ventricular outflow tract, and handmade expanded polytetrafluoroethylene (ePTFE)-valved conduits have been widely used instead. This study aimed to clarify the long-term outcomes and the durability of the ePTFE-valved conduits purely by narrowing down to those with large sizes to eliminate the influence of the body growth. METHODS Between January 2002 and December 2015, patients who underwent right ventricular outflow tract reconstruction in 34 Japanese institutions using ePTFE-valved conduits with a diameter of ≥18 mm were included. All the valved conduits were made in the authors’ institution and delivered to each participating institution. RESULTS Overall, 502 patients were included. Early mortality was 1.4% and not related to conduit failure. The overall survival rate was 98.2% at 5 years and 96.6% at 10 years. Freedom from conduit explantation was 99.5% at 5 years and 89.0% at 10 years. Three patients (0.13 per 100 patient-years) developed infective endocarditis of the conduit, and only 1 patient required conduit removal. Pulmonary insufficiency was mild or less in 480 (96%) patients, and conduit stenosis was mild or less in 436 (88%) patients at the latest follow-up. CONCLUSIONS By narrowing the analyses down to only ePTFE conduits with a large size, satisfactory long-term outcomes of these conduits with a fan-shaped valve and bulging sinuses were shown. These conduits would be among the optimal choices for right ventricular outflow tract reconstruction.

Recently, number of adult congenital heart disease(ACHD) patients are rapidly increasing, but statistical evidences in adult congenital heart surgery are not sufficient because of its short history in addition to the complicated pathophysiology of ACHD. Therefore, a "heart team" including surgeons, pediatricians, and cardiologists are required, and blood flow imaging and 4-dimensional (4D) imaging plays an essential role in perioperative patient management. Enhanced 3-dimensional computed tomography( 3D-CT) scan is useful to determine cardiopulmonary bypass strategies in reoperation, and 4D flow magnetic resonance imaging( MRI) is useful in assessment of complicated hemodynamics. Diastolic function of systemic ventricle is very important in right side heart valve surgery or univentricular heart surgery, echocardiography is essential to examine the valve structure, and electrophysiological mapping is useful in concomitant arrhythmia surgery. This article describes the indications, procedures, and perioperative managements of surgery in the right ventricular outflow tract, tricuspid valve, aortic root, Fontan circulation, and systemic right ventricle.

A 78-year-old woman underwent partial hepatectomy for the treatment of hepatocellular carcinoma(HCC).After surgery, she was further treated with radiofrequency ablation(RFA)and transcatheter arterial embolization(TAE)for the intrahepatic reccurence of HCC.Thirty months after surgery, her tumor marker levels increased and computed tomography (CT)revealed an intrahepatic mass and right renal tumor which extended into the renal vein.We therefore suspected a reccurence of HCC and right renal cell carcinoma which extended into the renal vein, and thus performed right nephrectomy before again performing TAE to treat the reccurence of HCC.The pathological findings showed HCC metastasis in the kidney. Nephrectomy should be performed as soon as possible in patients with a renal vein tumor thrombus in order to prevent the occurrence of a pulmonary embolism.We herein describe a very rare oncologic emergency case and review the relevant literature.

We report a case of an elderly patient diagnosed with intrahepatic cholangiocellular carcinoma with cholangitis.An 88- year-old woman presented with fever.Computed tomography examination showed a hepatic tumor and dilation of the peripheral bile duct.Cholangiocarcinoma was diagnosed.Biliary tract drainage and palliative care were suggested because of her age.However, her family asked for a second opinion about operative management and consulted our hospital.Radical operations for intrahepatic cholangiocellular carcinoma that has spread to the left lobe are usually accompanied by widespread lymph node dissection and extensive hepatectomy with biliary tract reconstruction.However, in this case, the patient was very elderly and was able to safely undergo an operation by reduction of the excision range and by omitting lymph node dissection.We conclude that appropriate surgery and postoperative care to prevent complications are necessary when performing very invasive surgery in elderly patients.

A 54-year-old man was presented at our hospital with weight loss.He diagnosed with colorectal cancer, multiple liver metastases and para-aortic lymph node metastasis.After undergoing colostomy, he was treated sequentially with mFOLFOX6 plus bevacizumab(Bmab), FOLFIRI plus Bmab or Pmab, according to the guideline.Since these chemotherapy resulted in progressive disease, regorafenib was administered as a salvage-line treatment.PET -CT showed only para-aortic lymph node swelling with high FDG uptake.Severe adverse effects were developed shortly after regorafenib treatment so he requireda reduction in dose.Three years after treatment with regorafenib, the response of the target lesion was stable disease according to the RECIST criteria.Tumor growth had been controlled for a long time.

Ten lactating cows were used to determine the effect of feeding non-fibrous carbohydrate (NFC) supplement before grazing on feed intake and nitrogen (N) utilization throughout a grazing season. The experiment was conducted from June to September. Cows grazed twice a day (2.5 h × 2) under a set stocking system and were fed NFC supplement (1 kg/4 kg of milk yield) 2 h before grazing (PRE) or immediately after grazing (POST). Cows were also fed a grass and corn silage mixture ad libitum. Herbage dry matter intake (DMI) was greater for PRE than for POST throughout the experiment and decreased from June to September. Conversely, silage DMI was less for PRE than for POST throughout the experiment and increased over the grazing season. Consequently, total DMI for PRE did not differ from that for POST. Milk urea-N concentration and urinary urea-N excretion in June did not differ between the treatments, whereas that after July was higher for PRE than for POST. Proportion of urinary N excretion to absorbed N intake in June was lower for PRE than for POST, but that after July was higher for PRE than for POST. Feeding NFC supplement before grazing would improve N utilization when cows eat large amounts of herbage high in N.

This study evaluated the suitability of easily digested fiber sources as a supplemental fiber to improve overall fiber digestion in ruminants. First, the degradation of five fibrous feedstuffs and the stimulatory effects on rumen bacteria were examined in situ. Chickpea and lablab bean husks were selected for their potential use due to their large degradable fraction (>94%), which had a stimulatory effect on fibrolytic rumen bacteria such as Fibrobacter succinogenes. Second, a possible improvement in the digestibility of rice straw diet by husk supplementation was monitored in vivo. Four dietary treatments comprising RS (rice straw and concentrate), CHM (RS supplemented with Myanmar chickpea husk), CHE (RS with Egyptian chickpea husk) and LH (RS with lablab bean husk) were allocated to four wethers. The digestibility of acid detergent fiber was 3.1-5.5% greater in CHM and LH than RS. Total volatile fatty acid concentration was higher in LH than other treatments. Acetate proportion was higher in LH than RS. Ruminal abundance of F. succinogenes was 1.3-1.5 times greater in CHM and LH than RS. These results suggest that bean husk supplementation, especially lablab bean husk, might improve the nutritive value of rice straw diet by stimulating fibrolytic bacteria.

Six Thoroughbred horses were used to determine the mean retention time (MRT) of digesta in the different segments of the hindgut. The horses were fed timothy hay or silage in equal amounts (1.6% bodyweight [BW]/day) every 3 h/day. Hay or silage labeled with the rare earth elements cerium, dysprosium, neodymium, ytterbium, lanthanum, samarium and praseodymium, were fed to the horses 36, 30, 24, 18, 12, 6 and 3 h before slaughter, respectively. Just after slaughter, digesta samples in the different segments were collected and determined for the contents of the rare earth elements. Using these data, a method to calculate the MRT in the different segments was proposed. There were no significant differences between diets in the MRT of each segment. The averaged MRT of hay and silage in the cecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon and small colon was 2.9, 3.1, 5.9, 1.0, 4.0, and 4.0 h, respectively. The dry matter weight of the digesta was related to the MRT in the right ventral colon (r = 0.94, P = 0.005), left ventral colon (r = 0.87, P = 0.03), left dorsal colon (r = 0.98, P

To obtain basic knowledge about selecting horses for therapeutic riding, the influence of equine conformation on rider oscillation and relationships between these factors and the evaluation on horses as the therapeutic riding were studied. Thirty-five riding horses were used. Equine conformation was estimated by 24 indices. Rider oscillation was measured by an accelerometer fixed at the rider’s waist. The spatial position of the oscillation was estimated by a double integration of the acceleration. Horses were evaluated for therapeutic riding by a Riding for the Disabled Association instructor as a rider. Evaluations were on a scale of 1 to 5, with 5 being the highest score for 27 items. Horses were classified into 4 groups: the short and narrow (SN), short and wide (SW), tall and narrow (TN), and tall and wide (TW). The frequencies of rider oscillation both at walk and trot were higher (P

Under rotational grazing by lactating dairy cows, effects of grass height at the start of a grazing season on herbage intake and milk production were evaluated. A 1.87 ha grass-clover mixed sward was divided evenly into two paddocks, and a different group of six lactating Holstein cows were rotationally grazed on each paddock for 5 h a day. Grazing was commenced when mean grass height in the two paddocks reached 15 cm (S15) and 20 cm (S20), respectively. Mean grass height throughout the grazing period tended to be higher in S20 than S15, and in particular, the difference at June between the treatments was significant (P

Four Holstein cows were used to determine the effect of timing of the feeding of a corn silage (CS)-based supplement on the feed intake, milk production and nitrogen utilization of grazing dairy cows. The cows were fed the supplement 2 h before grazing (pre-grazing) or immediately after grazing (post-grazing). Cows were grazed for 5 h per day under a rotational grazing system. There was no difference in the herbage and total feed intake between treatments. The milk protein yield for pre-grazing tended to be higher than that for post-grazing, whereas the milk yield did not differ between treatments. The total nitrogen intake for pre-grazing tended to be higher than that for post-grazing (P = 0.06). There was no difference in the urinary nitrogen output between treatments, whereas the proportion of urinary nitrogen output : total nitrogen intake for pre-grazing tended to be lower than that for post-grazing (P = 0.06). The milk nitrogen output and nitrogen retention for pre-grazing tended to be higher than that for post-grazing (milk nitrogen, P = 0.06; nitrogen retention, P = 0.05). Nitrogen utilization of grazing dairy cows was improved by feeding a CS-based supplement before grazing.

The effects of supplementary corn silage (CS) of either 2 or 4 kg of dry matter (DM; S + 2 and S + 4, respectively) above the energy requirement for milk production and maintenance for grazing dairy cows (S) were determined. Time-restricted grazing was used to compare the feed intake, milk production, and nitrogen and energy use of lactating cows. The experiment was carried out on two different pastures using a 3 × 3 Latin square design for each pasture. Cows were grazed for 5 h on a rotational grazing system and were fed concentrate (1 kg per 5 kg of milk yield). Herbage intake was measured using a weighing technique. To calculate the energy and nitrogen use, whole feces and urine were collected. There was no statistical effect of the pastures. Herbage intake decreased by the addition of CS (P = 0.02). The reduction of herbage DM intake per unit consumption of supplementary CS towards the S group were 0.80 and 0.45 kg for the S + 2 and S + 4 groups, respectively. The total DM intake for the S + 4 group was higher than that for the S and S + 2 groups (P = 0.02). Milk yield did not differ among treatments, even though the total DM intake for the S + 4 group was higher than that of the S and S + 2 groups. Nitrogen and energy use did not differ with the addition of CS.

To clarify the effects of sheep pasturing to meadow in open space on plant height, standing crop, and people's impression, six sheep were pastured in the grass (about 800m&sup2) in the campus of Hokkaido University for 13 days. Although the plant height or standing crop could be controlled by sheep grazing, the extent differed depending on the plant species. Influences of variability of the growth level of each plant species and the palatability of each plant species on the control of plants were considered. In further experiments, effects of the timing of pasturage or palatability on the control of plant height or standing crop should be confirmed. In the survey, it was clarified that respondents had regarded the sheep favorably or the scenery including sheep, and generally agreed with the use of sheep in grass management. However, there is a possibility that many of respondents have been interested in sheep pasturage in advance, because there were several reports by the media before pasturage. Attitudes of residents or tourists to the sheep grazing should be further evaluated.

A near-infrared (NIR) spectroscopic sensing system was constructed on an experimental basis. This system enables NIR spectra of unhomogenized milk to be obtained during milking over a wavelength range of 600 to 1050 nm. The NIR system can be used forreal-time monitoring of fat, protein, lactose, somatic cell count, and milk urea nitrogen during milking with sufficient precision and accuracy. By using this NI Rsystem, dairy farm management could proceed to the next step in the transition to artificial intelligence in dairy farming based on information of each individual cow.

Seventy-six animals including cattle, sheep, horses, 6 species of zoo animals were employed for collection of fresh feces in order to detect verotoxigenic Esherichia coli (VTEC) by safe, quick and sensitive PCR-based molecular methods. Bacterial cell disruption with bead-beating followed by bacterial DNA purification with hydroxyapatide chromatography and gel filtration allowed DNA preparation from animal feces with high recovery and purity. A mountain goat was firstly shown by PCR and seqiencing to shed verotoxin 2 gene (vt2) that was used to generate vt2 probe and second primer set for nested PCR to attempt more sensitive detection. Most sensitive nested PCR revealed that 45% of tested cattle and 47% of tested zoo animals were VTEC-positive, while least sensitive normal PCR detected VTEC from none of these animals except a mountain goat. Moderately sensitive detection by PCR in combination with hybridization suggested that the VTEC density varied between the VTEC-positive cattle.

北海道和種馬に同一原料牧草（チモシー）から調製した無細切の乾草およびサイレージを乾物で同量（2.2％BW）給与し，消化率，咀嚼活動，糞の粒度分布および消化管内容物の平均滞留時間を比較検討した．乾物および繊維成分の消化率には飼料間で差はなかった．乾物摂取量あたりの咀嚼回数は乾草よりサイレージが多かったが（4.6 vs. 5.3回/gDM, P＜0.05），糞の粒度分布は乾草摂取時よりもサイレージ摂取時が大粒子画分（2.36mm以上）の割合が高く（29 vs. 51％, P＜0.05），小粒子画分（0.30mm以上1.18mm未満）の割合が低かった（35 vs. 23％, P＜0.05）．消化管内容物の平均滞留時間は，乾草よりもサイレージが，液相（21.1 vs. 16.9h, P＜0.01）および飼料全体（23.7 vs. 21.8h, P＜0.05）ともに短く，また両飼料とも液相よりも飼料全体が長かった（P＜0.01）．葉部および茎部の滞留時間には両飼料とも差がなかった．

Fractal analysis was applied to grazing paths in non-patchy environments to clarify the behavioral information available from the analysis and to evaluate the effect of social factors on grazing paths. The grazing paths of Holstein Friesian cows in groups of 4 (GROUP-4) and 10 cows (GROUP-10) on a spatially homogeneous pasture were observed. Application of fractal analysis could quantify three aspects of grazing paths of cows: (1) hierarchical scales, (2) structure and (3) sinuosity by scale. In this study, regardless of the group size, the hierarchical scale of grazing paths was approximately 10 m. The paths showed different patterns below and above the 10 m scale. For both group sizes, fractal dimension (an index of sinuosity) below the 10 m scale was smaller than fractal dimension above 10 m scale suggesting that the paths below the 10 m scale were straighter than that above the 10 m scale. The fractal dimensions below and above the 10 m scale were significantly larger for GROUP-4 than for GROUP-10 (P0.05). Sinuosity of paths of GROUP-4 was higher than that of GROUP-10 over the entire scale. From these three aspects, fractal analysis can clarify the factors related to grazing paths of herbivores.

Ten lactating cows were used to determine the effect of feeding non-fibrous carbohydrate (NFC) supplement before grazing on feed intake and nitrogen (N) utilization throughout a grazing season. The experiment was conducted from June to September. Cows grazed twice a day (2.5 h × 2) under a set stocking system and were fed NFC supplement (1 kg/4 kg of milk yield) 2 h before grazing (PRE) or immediately after grazing (POST). Cows were also fed a grass and corn silage mixture ad libitum. Herbage dry matter intake (DMI) was greater for PRE than for POST throughout the experiment and decreased from June to September. Conversely, silage DMI was less for PRE than for POST throughout the experiment and increased over the grazing season. Consequently, total DMI for PRE did not differ from that for POST. Milk urea-N concentration and urinary urea-N excretion in June did not differ between the treatments, whereas that after July was higher for PRE than for POST. Proportion of urinary N excretion to absorbed N intake in June was lower for PRE than for POST, but that after July was higher for PRE than for POST. Feeding NFC supplement before grazing would improve N utilization when cows eat large amounts of herbage high in N.

This study evaluated the suitability of easily digested fiber sources as a supplemental fiber to improve overall fiber digestion in ruminants. First, the degradation of five fibrous feedstuffs and the stimulatory effects on rumen bacteria were examined in situ. Chickpea and lablab bean husks were selected for their potential use due to their large degradable fraction (94%), which had a stimulatory effect on fibrolytic rumen bacteria such as Fibrobacter succinogenes. Second, a possible improvement in the digestibility of rice straw diet by husk supplementation was monitored in vivo. Four dietary treatments comprising RS (rice straw and concentrate), CHM (RS supplemented with Myanmar chickpea husk), CHE (RS with Egyptian chickpea husk) and LH (RS with lablab bean husk) were allocated to four wethers. The digestibility of acid detergent fiber was 3.1-5.5% greater in CHM and LH than RS. Total volatile fatty acid concentration was higher in LH than other treatments. Acetate proportion was higher in LH than RS. Ruminal abundance of F. succinogenes was 1.3-1.5 times greater in CHM and LH than RS. These results suggest that bean husk supplementation, especially lablab bean husk, might improve the nutritive value of rice straw diet by stimulating fibrolytic bacteria.

Cetaceans are well adapted to their hyperosmotic environment by properly developed osmoregulatory ability. A question here is how they regulate water and mineral balances in marine habitats. In the present study, we determined blood and urine levels of various chemicals involved in osmoregulation, compared them with those in artiodactyls, and characterized the values in the whales. Blood and urine samples obtained from baleen whales of common minke (Balaenoptera acutorostrata), sei (B. borealis), and Bryde's whales (B. brydei), and toothed whales of sperm whales (Physeter macrocephalus) were analyzed for osmolality, major electrolytes, urea, steroid hormones and glucose. The urine osmolality and Na(+) concentrations in the cetaceans were much higher than those in the cattle. Furthermore, the cetaceans had 5 to 11-fold urea in plasma than the cattle, and 2 to 4-fold urea in urine. There were no significant difference in the plasma concentrations of corticosteroids between the cetaceans and the cattle. The present results indicate that the osmoregulatory parameters seem to be not affected by the reproductive stage and sex steroid hormones. The concentrations of urea in plasma and urine of the baleen whales were higher than those of the sperm whales, indicating a possibility that their osmoregulatory mechanisms may be correlated to their feeding habits. The present results suggest that cetaceans have unique osmoregulatory mechanisms by which they excrete strongly hypertonic urine to maintain fluid homeostasis in marine habitats.

Written for presentation at the 2005 ASAE Annual International Meeting Sponsored by ASAE, Recently, there has been a need by dairy farmers for a method to assess milk quality in real time during milking. We have constructed an on-line near-infrared (NIR) spectroscopic sensing system on an experimental basis. The NIR system can be used for real-time monitoring of milk quality items such as fat, protein, lactose, somatic cell count, and milk urea nitrogen during milking with sufficient precision and accuracy. We tried to improve the robustness of calibration (CAL) models for measurement of milk quality items using NIR spectrum data obtained from two dairy herds. When CAL models developed from data obtained from one herd were used for validation of data obtained from the same herd, the milk quality items could be measured with high levels of accuracy. On the other hand, when the CAL models were used for validation of data obtained from the other herd, the levels of accuracy in measurements of all milk quality items except fat were low. The low levels of accuracy may be caused by factors such as differences in cow individuality, lactation stage, calving times, feeding stage and experimental period (temperatures in the dairy barn). To develop robust calibration models for measurement of milk quality items, therefore, data acquisition from various milk spectra caused by these factors is necessary.

The hepatic synthesis of acute phase proteins in ruminants has been suggested to be regulated by some mechanisms different from those in other species such as rodents and human. To explore possible regulatory factors unique to ruminants, we examined effects of interleukin (IL)-6, IL-1 and tumor necrosis factor (TNF), on haptoglobin (Hp) synthesis using a primary culture system of bovine hepatocytes. After bovine primary cultured hepatocytes were incubated in the presence of various concentrations of the cytokines, the synthesis and mRNA level of haptoglobin and albumin were measured by labeling with [35S]-methionine and immunoprecipitation, and by Northern blot analysis, respectively. Hp synthesis was dose-dependently increased by recombinant human (rh) IL-6, and also by rhTNF-alpha, but to a less extent, while it was not affected by rhIL-1 beta. The stimulatory effect is mainly pretranslational, because mRNA level of Hp changed in parallel with protein synthesis. In contrast, albumin synthesis was suppressed by these three cytokines similarly. These results are inconsistent with the previously proposed view that TNF and IL-1 overlap in their pathways leading to the transcriptional activation of many acute phase protein genes. In conclusion, there is a species-specific unique signaling system, especially for TNF, in transcriptional activation of bovine Hp gene.

We describe a technique for isolation and primary culture of bovine hepatocytes, and their metabolic characterization. Hepatocytes were isolated from the caudate lobe of bovine liver by perfusion with 0.25 mM ethylene-glycol tetraacetic acid and 0.05% collagenase. The viability and yield of the cells were 70-92% and 0.1-3.6 x 10(7) cells/g liver, respectively. When the isolated hepatocytes were cultured in Williams' medium E, they began to spread in 3 hr and formed monolayers in 24 hr. These monolayers were retained for at least 6 days. To monitor the metabolic activities specific to liver, synthesis and secretion of albumin were measured by labeling with [35S]-methionine and immunoprecipitation. This activity was low in isolated hepatocytes, but increased after culturing 1-3 days, and decreased again after 6 days. Glycogenolytic activity was also assessed by measuring glucose release to the medium by stimulation with epinephrine. The glycogenolytic response to epinephrine was also enhanced by culturing the hepatocytes 1-3 days, but was decreased after 6 days. Since the isolated bovine hepatocytes retained the liver-specific activities of albumin synthesis and glycogenolysis for several days in culture, these cells are useful for cellular and molecular studies on the functions of bovine liver.