Several Amino Acids and Carnitine Transport Activities of the Epithelial Cells of Bovine Mammary Gland

We investigated several amino acids and carnitine transport activities of bovine mammary gland epithelial cells. Gly, Ala, Gln, Glu, Arg, Leu, cystine and carnitine transport activities at 1 μmol/L substrate concentration were 24.0 ± 3.97, 90.9 ± 13.4, 32.5 ± 9.0, 14.2 ± 5.1, 48.9 ± 11.4, 48.8 ± 5.1, 22.7 ± 6.8 and 2.56 ± 0.96 nmol/mg protein/min, respectively. Na-dependency of transport was observed in Gly, Ala and Gln, but not in Arg, Leu and carnitine. Glu and Cys transport activity without Na condition were reduced to 36%, 63% in Na-free condition, respectively. Carnitine transport activities were low but detectable with or without Na condition. There was no correlation between amino acid transport activities and their concentrations in milk. The data clarified in this paper will be basic data for metabolic analysis of bovine mammary gland

Case report: Abdominal hernia repair using a surgical wire and an autologous omental graft in a Japanese Black calf.

BACKGROUND Hernia formation is a well-known abdominal wall disorder in calves and most often occurs in the umbilical region. In addition, it occurs in the abdominal wall outside the umbilical region. It has been reported to involve acquired factors, such as external force to the lower or lateral abdominal wall, trauma, muscle weakness, and, although rare, congenital or hereditary factors. Although there have been reports on the repair of abdominal wall hernias caused by abnormal muscle formation, there have been no reports on the treatment of abdominal wall hernias caused by abnormal vertebral and rib formation in cattle or other ruminants. In this study, for the first time, we describe a case of successful closure of a hernia in the left flank caused by malformation of the vertebrae and ribs. The repair was performed by narrowing the ribs using a surgical wire and covering the defect with a free omental graft. CASE PRESENTATION A male Japanese Black calf showed a mild bulge of the left abdominal wall and abnormal morphology of the left ribs immediately after birth. At 3 months of age, computed tomography revealed fusion of the 9th, 10th, and 11th thoracic vertebrae and missing formation of the 10th and 11th left ribs at the thoracic vertebral fusion site. Additionally, a 15.2 × 24.4 cm abdominal hernia had formed in the abdominal wall. During surgery, the ribs forming the hernia were narrowed with a surgical wire, and the hernia was covered with a large autologous free omental graft to prevent intra-abdominal organ prolapse. At the monthly follow-up (11 months after surgery), the calf had developed similarly to other calves of the same age on the farm, and no complications were noted. CONCLUSIONS The current case shows that a combination of rib correction using a surgical wire combined with transplantation of a free autologous greater omentum graft was shown to be a possible treatment option for surgical repair of large abdominal wall hernias caused by rib malformation

Amino Acid Transport System N: Molecular Structure, Distribution and Functional Analysis of Canine SLC38A5 (SNAT5) in Lens Epithelial Cells.

Na-dependent of neutral amino acid transport activity in canine lens epithelial cells (LEC) line was investigated. The transporter activity of glutamine was 11.17 ± 3.17 nmol/mg protein/min, and it was reduced by 75% in the absence of sodium. The full-length cDNA sequence of canine sodium-dependent neutral amino acid transporter 5 (SNAT5) was 2151 bp long and was predicted to encode the 536 amino acid polypeptides. The deduced amino acid sequence of canine SNAT5 showed >80% similarities with those of human and mouse. The RT-PCR analysis indicated that SNAT5 was expressed in liver, kidney and LEC, but not in heart and skin

Visualizing bovine leukemia virus (BLV)-infected cells and measuring BLV proviral loads in the milk of BLV seropositive dams

International audienceAbstractBovine leukemia virus (BLV) infects cattle and causes serious problems for the cattle industry, worldwide. Vertical transmission of BLV occurs via in utero infection and ingestion of infected milk and colostrum. The aim of this study was to clarify whether milk is a risk factor in BLV transmission by quantifying proviral loads in milk and visualizing the infectivity of milk. We collected blood and milk from 48 dams (46 BLV seropositive dams and 2 seronegative dams) from seven farms in Japan and detected the BLV provirus in 43 blood samples (89.6%) but only 22 milk samples (45.8%) using BLV-CoCoMo-qPCR-2. Although the proviral loads in the milk tended to be lower, a positive correlation was firstly found between the proviral loads with blood and milk. Furthermore, the infectivity of milk cells with BLV was visualized ex vivo using a luminescence syncytium induction assay (LuSIA) based on CC81-GREMG cells, which form syncytia expressing enhanced green fluorescent protein (EGFP) in response to BLV Tax and Env expressions when co-cultured with BLV-infected cells. Interestingly, in addition to one BLV-infected dam with lymphoma, syncytia with EGFP fluorescence were observed in milk cells from six BLV-infected, but healthy, dams by an improved LuSIA, which was optimized for milk cells. This is the first report demonstrating the infectious capacity of cells in milk from BLV-infected dams by visualization of BLV infection ex vivo. Thus, our results suggest that milk is a potential risk factor for BLV vertical spread through cell to cell transmission

イヌのグルコース輸送体4（GluT4）の分子構造と組織発現

イヌのグルコース輸送体4（GluT4）の分子構造と組織発現を調べた。イヌGLUT4のcDNA配列全長1637bpであり，510のアミノ酸から構成され，ヒトとマウスより1アミノ酸大きいことが明らかとなった。このアミノ酸はヒトGluT4と95.2％，マウスGluT4と93.7％の相同性を示した。RT-PCRの結果，心臓・骨格筋・脂肪組織に発現がみられた。ウェスタンブロット解析により，GluT4タンパクは50kDaの質量をもつことが示された。イヌのインシュリン感受性グルコース輸送体4の構造が判明したことにより，イヌの糖尿病の病態生化学の理解に寄与するものと考えられる。Molecular structure and distribution of canine glucose transporter 4(GluT4)　were investigated. Canine GluT4 was consisted of 1637 nucleotides encoding 510 amino acids, which is one amino acid longer than human and mouse. Canine GluT4 exhibited 95.2% and 93.7% identity to human and mouse, respectively RT-PCR analysis indicated that its expression was confirmed in heart, skeletal muscle and lipocytes. Westernblot analysis showed the molecular weight of canine GluT4 was 50kDa. Molecular structure and distribution of insulin-sensitive GluT4 will contribute to a better understanding of the patho-physiology of canine diabetes

Arthroscopic approaches to and anatomy of the shoulder joint of cattle: a cadaver study.

BACKGROUND Arthroscopic surgery is described as a minimally invasive technique for diagnosis, exploration and treatment of joint disorders. It allows intraarticular structures to be assessed accurately, thereby improving the diagnostic capabilities, and it broadens the spectrum of surgical techniques feasible for treatment of articular pathologies in cattle. This study aimed to assess for cattle the described arthroscopic approaches to the shoulder joint of horses, and to describe the appearance of the corresponding intraarticular structures of the shoulder joint. Additionally, to perform histological examination where tissues were identified and assessed arthroscopically, but the tissue type was uncertain using cadaveric limbs from cattle of different age categories without any signs of orthopedic diseases of the front limbs. RESULTS An anatomic and arthroscopic investigation with 34-cadaveric forelimbs from 20-cattle was performed. The arthroscope was inserted either immediately cranial or 1-cm caudal to the tendon of the infraspinatus muscle for the cranial and caudal approaches, respectively. The shoulder joints were examined with the limbs in either horizontal non-pulled position, abducted non-pulled position using a three-pod limb holder adjustable in height, or horizontal manually pulled position. Arthroscopy was performed using a rigid 30°arthroscope (18-cm length, 4-mm outer diameter) to view the synovial pouches with their synovial villi and the following structures: cranial rim of the glenoid, cranial portion of the humeral head, incisura-glenoidalis, caudal rim of the glenoid, caudal portion of the humeral head, and cranial and caudal cul-de-sac. Abduction of the limb allowed improved visualization of the lateral portion of the joint. Pulling the limb facilitated investigation of the medial portion of the joint. Generally, the distention range was higher in younger as compared to adult cattle, and visualization of the medial portion of the joint was, therefore, facilitated in younger animals. The main complications observed were subcutaneous fluid extravasations and partial-thickness articular cartilages wear-lines. CONCLUSION The described arthroscopic techniques allowed good overall visualization of the most relevant anatomical structures within the healthy cadaveric joint. Further investigations are warranted to evaluate the diagnostic and therapeutic applications of these techniques and the prognosis of arthroscopic surgery as a tool for the treatment of joint lesions

Comparison of Vaccination Protocols for Bovine Herpesvirus Type1 and Bovine Viral Diarrhea Virus

We investigated the immune responses against bovine herpesvirus type 1 (BHV-1) and bovine viral diarrhea virus (BVDV) by two different vaccination protocols. An attenuated-live vaccine (containing BHV-1 and BVDV-1) and an inactivated vaccine (containing inactivated virus antigens of BHV-1, BVDV-1, and BVDV-2) were used. Two different immunization protocols were investigated: inoculation of live vaccine 28 days after inactivated vaccine inoculation (KL), and inoculation of inactivated vaccine 28 days after live vaccine inoculation (LK). Antibodies against BHV-1, BVDV-1, and BVDV-2 were examined. Antibody titer against BHV-1 was significantly higher in calves vaccinated by the KL protocol 30 days post inoculation (dpi). On the other hand, antibody titer against BVDV-1 was significantly lower in calves vaccinated by the KL protocol 30 dpi. However, approximately equivalent antibody titers were observed using either protocol by 56 dpi. No significant difference in antibody titer against BVDV-2 was observed between the two protocols, with a nearly equivalent immune response acquired by 56 dpi. These results suggest that when combination vaccines are used, the vaccination protocol should be selected depending on the prevalence of infectious diseases in each farm