<p>The approved Japanese measurement method of circulating alkaline phosphatase (ALP) has changed from that of the Japan Society of Clinical Chemistry (JSCC) to that of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). We measured the serum levels of total ALP (t-ALP) and those of the isoenzymes ALP2 and ALP3 in 50 Asian elephant (<i>Elephas maximus</i>) specimens using both methods. The activities determined by the IFCC method were roughly one-third lower than those determined by the JSCC method. We present conversion formulae. Our results enable comparisons of historical and current data on serum ALP activities in endangered, zoo-managed Asian elephants.</p>

DOI： 10.1292/jvms.22-0440

PubMed

<p>This study aimed to elucidate the distribution of enrofloxacin (ERFX) within the bronchoalveolar region of pigs. Six clinically healthy pigs were allocated to intramuscular treatments with either a single dose of 5 mg/kg or 7.5 mg/kg ERFX. Samples of plasma and bronchoalveolar lavage fluid (BALF) were obtained from each pig 0 (before administration), 3, 8, and 24 hr after ERFX administration. The ERFX concentrations in pulmonary epithelial lining fluid (ELF) and BALF cells showed a similar pattern during the experimental period, whereby ERFX concentrations in both ELF and BALF cells were higher than those in the plasma. These results suggest that intramuscularly injected ERFX is well-distributed in the bronchoalveolar region.</p>

DOI： 10.1292/jvms.22-0416

PubMed

<p>Serum levels of creatine kinase (CK) and lactate dehydrogenase (LDH) isoenzymes were evaluated in nine zoo-managed Asian elephants (<i>Elephas maximus</i>) using a commercial agarose gel electrophoresis (AGE) kit. CK was separated into two major fractions, CK-BB and CK-MM, along with a small fraction of macroenzyme-CK type 2 (mCK2); CK-MM was the largest fraction. LDH was separated into five fractions (LDH1–5); LDH3 was the largest fraction. Age was negatively and positively correlated with the percentages of CK-BB and CK-MM, respectively, and negatively correlated with CK-BB and mCK2 activities. These results indicate that an AGE kit can be used to evaluate CK and LDH isoenzymes. Routine isoenzyme testing may enable early detection of disease and physiological changes.</p>

DOI： 10.1292/jvms.22-0550

PubMed

Publishing type：Research paper (scientific journal)   International / domestic magazine：Domestic journal  

Understanding the immune dynamics in the respiratory mucosa of calves is necessary for a good management of bovine respiratory disease. Immune dynamics in the respiratory mucosa in humans and experimental animals has been assessed by flow cytometric analysis of bronchoalveolar lavage fluid (BALF); however, few reports have addressed this subject in calves. The aim of this study was to establish a universal method to analyze bronchoalveolar lavage fluid (BALF) by flow cytometry and to obtain basic knowledge of bovine respiratory mucosal immune dynamics. We investigated the immune cell populations in BALF and evaluated the surface antigen expression of alveolar macrophages in calves using flow cytometer. To further analyze the surface antigen variation observed in alveolar macrophages in detail, stimulation assays were performed in vitro. BALF cells were separated into three distinct populations based on their light scatter plot, which were considered to be macrophages, lymphocytes, and neutrophils. In most individuals, most of the BALF immune cells were alveolar macrophages, but an increased proportion of lymphocytes and neutrophils was observed in some individuals. Analysis of each surface antigen expression in alveolar macrophages showed that CD21 and MHC class II expression changed in response to changes in the leukocyte population. Moreover, when alveolar macrophages were stimulated with interferon-γ in vitro, the expression of CD21 was drastically reduced and MHC class II was increased, suggesting that functional changes in alveolar macrophages themselves are involved in the immune dynamics.

DOI： 10.1292/jvms.21-0522

PubMed

Publishing type：Research paper (scientific journal)   International / domestic magazine：Domestic journal  

This study analyzed the pharmacokinetics of orbifloxacin (OBFX) in plasma, and its migration and retention in epithelial lining fluid (ELF) and alveolar cells within the bronchoalveolar lavage fluid (BALF). Four healthy calves received a single dose of OBFX (5.0 mg/kg) intramuscularly. Post-administration OBFX dynamics were in accordance with a non-compartment model, including the absorption phase. The maximum concentration (Cmax) of plasma OBFX was 2.2 ± 0.1 μg/ml at 2.3 ± 0.5 hr post administration and gradually decreased to 0.3 ± 0.2 μg/ml at 24 hr following administration. The Cmax of ELF OBFX was 9.3 ± 0.4 μg/ml at 3.0 ± 2.0 hr post administration and gradually decreased to 1.2 ± 0.1 μg/ml at 24 hr following administration. The Cmax of alveolar cells OBFX was 9.3 ± 2.9 μg/ml at 4.0 hr post administration and gradually decreased to 1.1 ± 0.2 μg/ml at 24 hr following administration. The half-life of OBFX in plasma, ELF, and alveolar cells were 6.9 ± 2.2, 7.0 ± 0.6, and 7.8 ± 1.6 hr, respectively. The Cmax and the area under the concentration-time curve for 0-24 hr with OBFX were significantly higher in ELF and alveolar cells than in plasma (P<0.05). These results suggest that OBFX is distributed and retained at high concentrations in ELF and alveolar cells at 24 hr following administration. Hence, a single intramuscular dose of OBFX (5.0 mg/kg) may be an effective therapeutic agent against pneumonia.

DOI： 10.1292/jvms.21-0517

PubMed

Publishing type：Research paper (scientific journal)   International / domestic magazine：International journal  

To understand the pathogenesis of bovine respiratory disease (BRD), it is necessary to elucidate the mechanisms of alveolar macrophage regulation by cytokines and pathogen-associated molecular patterns (PAMPs). Moreover, "non-specific effects (NSEs)" an innate immune regulatory mechanism in response to vaccines containing PAMPs, has recently attracted attention. It may be applied to BRD control, but there is limited knowledge in bovine. To investigate this, we stimulated alveolar macrophages in vitro with lipopolysaccharide (LPS), polyinosinic-polycytidylic acid sodium salt (Poly I:C), interferon gamma (IFN-γ), and modified-live viral (MLV) vaccines, respectively, and analyzed changes in tumor necrosis factor alpha (TNF-α), inducible nitric oxide synthase (iNOS), and interferon beta (IFN-β) mRNA expression levels. mRNA expression levels of TNF-α, iNOS, and IFN-β were significantly increased in bovine alveolar macrophages stimulated by IFN-γ and MLV vaccine; LPS, IFN-γ, and MLV vaccine; and MLV vaccine only, respectively. Additionally, all MLV vaccine-stimulated mRNA expression increases were observed in a concentration-dependent manner. These results revealed in part, the mechanism of bovine alveolar macrophage regulation by cytokines and PAMPs. Understanding the regulatory mechanisms of alveolar macrophages will contribute to understanding the pathogenesis of BRD and preventive and therapeutic BRD management based on NSEs.

DOI： 10.1016/j.vetimm.2021.110378

PubMed

Publishing type：Research paper (scientific journal)   International / domestic magazine：International journal  

BACKGROUND: Medical interventions for subchondral bone cysts in horses have been extensively studied. This study investigated the regeneration of articular cartilage and subchondral bone with scaffold-free three-dimensional (3D) constructs of equine synovial membrane-derived mesenchymal stem cells (SM-MSCs) isolated from three ponies and expanded until over 1.0 × 107 cells at passage 2 (P2). RESULTS: SM-MSCs were strongly positive for CD11a/CD18, CD44, and major histocompatibility complex (MHC) class I; moderately positive for CD90, CD105, and MHC class II; and negative for CD34 and CD45 on flow cytometry and differentiated into osteogenic, chondrogenic, and adipogenic lineages in the tri-lineage differentiation assay. After culturing SM-MSCs until P3, we prepared a construct (diameter, 6.3 mm; height, 5.0 mm) comprising approximately 1920 spheroids containing 3.0 × 104 cells each. This construct was confirmed to be positive for type I collagen and negative for type II collagen, Alcian blue, and Safranin-O upon histological analysis and was subsequently implanted into an osteochondral defect (diameter, 6.8 mm; depth, 5.0 mm) at the right femoral medial condyle. The contralateral (left femoral) defect served as the control. At 3 and 6 months after surgery, the radiolucent volume (RV, mm3) of the defects was calculated based on multiplanar reconstruction of computed tomography (CT) images. Magnetic resonance (MR) images were evaluated using a modified two-dimensional MR observation of cartilage repair tissue (MOCART) grading system, while macroscopic (gross) and microscopic histological characteristics were scored according to the International Cartilage Repair Society (ICRS) scale. Compared to the control sites, the implanted defects showed lower RV percentages, better total MOCART scores, higher average gross scores, and higher average histological scores. CONCLUSIONS: Implantation of a scaffold-free 3D-construct of SM-MSCs into an osteochondral defect could regenerate the original structure of the cartilage and subchondral bone over 6 months post-surgery in horses, indicating the potential of this technique in treating equine subchondral bone cysts.

DOI： 10.1186/s12917-021-03126-y

PubMed

We investigated bovine respiratory disease complex by collecting bronchoalveolar lavage fluid (BALF) for bacteriological and virological examinations from fattening cattle following their introduction from market, and monitored their treatment records for any subsequent history of medical interventions. A cohort of 10 steers underwent BALF collection in April 2016, and <i>Pasteurella multocida</i> and <i>Mycoplasma bovis</i> were isolated in samples from eight and one animal, respectively, in bacteriology, and <i>Respiratory Syncytial</i> (RS) virus genetic material was detected in a sample from one animal in virology. A second cohort of 10 steers from the same herd underwent BALF collection in December 2016, and <i>Pasteurella multocida</i> and <i>Mycoplasma bovis</i> were isolated in samples from four and one animal, respectively, in bacteriology, and RS virus genetic material was detected in samples from all 10 animals in virology. Monitoring post-introduction treatment showed that the cohort with ubiquitously detected RS virus tended to require earlier medical interventions. These findings indicate the presence of pneumonia-causing bacteria or viruses in the bronchoalveolar region of cattle after introduction, which may be important for the development of preventive measures and treatment strategies.

DOI： 10.12935/jvma.74.497

CiNii Article

Other URL： http://id.ndl.go.jp/bib/031657317

市場導入肥育素牛から気管支肺胞洗浄液(BALF)を採取し、微生物学的に検査するとともに、供試牛の導入後治療状況を調査した。BALFからの細菌分離検査では、Pasteurella multocida及びMycoplasma bovisが分離された。また、ウイルス遺伝子検査では牛Respiratory syncytial(RS)ウイルス遺伝子が検出された。供試牛の導入後治療状況を調査した結果、牛RSウイルス遺伝子が全頭に確認された牛群では導入後に牛呼吸器病症候群治療を受けるまでの期間が早かった。これらの知見は、導入後の牛の気管支肺胞領域に肺炎原因菌あるいは原因ウイルスが存在することを示しており、予防としてのワクチネーションの徹底や使用する抗菌剤の的確な選定を含め、治療方針の決定に重要であると考えられた。(著者抄録)

Publishing type：Research paper (scientific journal)   International / domestic magazine：International journal  

A 27-y-old Anglo-Arabian gelding with bay coat color was presented with a swelling of the left maxillary region. Fenestration on the left maxilla revealed that the left maxillary sinus was filled with black-red tissue. A portion of the tissue was excised and diagnosed histologically as malignant melanoma. Genotyping of the STX17 gene for gray coat color revealed that the horse did not have the "gray" factor. The horse was euthanized ~3 mo after first presentation. During autopsy, a black-to-gray mass extended from the left nasal cavity to the surrounding paranasal sinus and invaded the hard palate, cribriform plate, and the cranial portion of the left olfactory bulb. Moreover, identical black nodules were present in lymph nodes from the mandible to the larynx, and in the spleen, liver, kidney, and adrenal glands. However, masses were not found in the skin, perineal region, or pelvic cavity. All of the black-to-gray nodules were malignant melanomas that were histologically identical to the initial biopsy; tumor emboli were also found in the kidney. Sinonasal mucosal melanoma is a rare disease in horses.

DOI： 10.1177/1040638720978561

PubMed

Kind of work：Joint Work  

Repository URL： http://hdl.handle.net/10466/00017577

Publishing type：Research paper (scientific journal)   International / domestic magazine：Domestic journal  

This study aimed to analyze the pharmacokinetics of enrofloxacin (ERFX) and its metabolite ciprofloxacin (CPFX) in plasma, as well as their migration to, and retention in, the epithelial lining fluid (ELF) and alveolar cells within the bronchoalveolar fluid (BALF). Four healthy calves were subcutaneously administered a single dose of ERFX (5 mg/kg). ERFX and CPFX dynamics post-administration were analyzed via a non-compartment model, including the absorption phase. The Cmax of plasma ERFX was 1.6 ± 0.4 µg/ml at 2.3 ± 0.5 hr post-administration and gradually decreased to 0.14 ± 0.03 µg/ml at 24 hr following administration. The mean residence time between 0 and 24 hr (MRT0-24) in plasma was 6.9 ± 1.0 hr. ERFX concentrations in ELF and alveolar cells peaked at 3.0 ± 2.0 hr and 4.0 ± 2.3 hr following administration, respectively, and gradually decreased to 0.9 ± 0.8 µg/ml and 0.8 ± 0.5 µg/ml thereafter. The plasma half-life (t1/2) of ERFX was 6.5 ± 0.7 hr, while that in ELF and alveolar cells was 6.5 ± 3.6 and 7.4 ± 4.3 hr, respectively. The Cmax and the area under the concentration-time curve for 0-24 hr for ERFX were significantly higher in alveolar cells than in plasma (P<0.05). These results suggest that ERFX is distributed at high concentrations in ELF and is retained at high concentrations in alveolar cells after 24 hr in the BALF region; hence, ERFX may be an effective therapeutic agent against pneumonia.

DOI： 10.1292/jvms.20-0110

PubMed

Publishing type：Research paper (scientific journal)   International / domestic magazine：Domestic journal  

The purpose of this study was to clarify the distribution of marbofloxacin (MBFX) within the bronchoalveolar region of pigs. Four clinically healthy pigs were intramuscularly injected with a single dose of MBFX (2 mg/kg). Samples of plasma and bronchoalveolar lavage fluid (BALF) were obtained for each pig at 0 (before administration), 3, 8 and 24 hr after administration of MBFX. As a result, the MBFX concentrations in pulmonary epithelial lining fluid (ELF) and in alveolar cells showed a similar pattern of concentrations during the experimental period. The MBFX concentrations both in ELF and alveolar cells were higher than in plasma. These results suggest that intramuscularly injected MBFX was well distributed in the bronchoalveolar region.

DOI： 10.1292/jvms.20-0239

PubMed

2週齢の黒毛和種仔ウシ(n=11)にβカロテン20mg/日を8週齢まで経口投与し、末梢血白血球への影響を調べた。投与群の血清βカロテン濃度は、4、8、12週齢時に非投与群に比べ有意に高値を示した。血清レチノール濃度に群間差はなく、2週齢時から12週齢時まで両群とも類似の漸増パターンを示した。このことから、βカロテン20mg/日を黒毛和種仔ウシに投与しても、血清レチノール濃度に大きな影響はないと考えられた。末梢血の総白血球数、単核球数、顆粒球数、およびCD3陽性(+)、CD8+、CD14+、CD21+、CD335+細胞数に有意な群間差はみられず、有意な経時変化も認められなかった。末梢血CD4+細胞数は、非投与群と比較して、投与群で4週齢時に有意に高値を示した。黒毛和種仔ウシにおいて、βカロテンの経口投与によって血清βカロテン濃度が上昇することで、末梢血の免疫担当細胞数が変化すると考えられ、βカロテン投与は末梢血白血球組成に影響を及ぼす可能性が示唆された。

In this study, nasopharyngeal swabs (Swab) and bronchoalveolar lavage fluids (BALF) were collected from 50 Japanese black cattle diagnosed with severe chronic pneumonia, and identification of the causative bacteria of pneumonia and its antibiotic susceptibilities were investigated. <i>Mycoplasma bovis</i> and <i>Pasteurella multocida</i> were mainly isolated from Swab and BALF. However, the rates at which the same bacterial species were isolated from both specimens in the same cow were relatively low. In addition, <i>M. bovis</i> and <i>P. multocida</i> isolated from BALF were mainly sensitive to fluoroquinolones, but the antibiotic susceptibilities of the same two species isolated from Swab were low. From the above, <i>M. bovis</i> and <i>P. multocida</i> were mainly isolated from the nasopharyngeal area and bronchoalveolar area of cattle with severe chronic pneumonia, but the antibiotic susceptibility differs depending on the collection site, even if the same bacterial species is a possibility. It seems necessary to be careful about estimating the pneumonia causing bacteria by Swab in calves with severe pneumonia.

DOI： 10.12935/jvma.73.31

CiNii Article

Other URL： http://id.ndl.go.jp/bib/030169943

本研究では、臨床的に重症慢性肺炎と診断された黒毛和種牛50頭から鼻咽頭スワブ(Swab)及び気管支肺胞洗浄液(BALF)を採取し、細菌分離とともに、その薬剤感受性を調査した。Swab及びBALFからはおもにMycoplasma bovis及びPasteurella multocidaが分離されたが、同一牛において両検体から同一細菌種が分離された割合は比較的低かった。また、BALFから分離されたM.bovis及びP.multocidaは、おもにフルオロキノロン系抗菌薬に感受性であったが、Swabから分離された同2菌種の同系抗菌薬に対する薬剤感受性は低かった。以上より、重症慢性肺炎罹患牛の鼻咽頭領域及び気管支肺胞領域からは、おもにM.bovis及びP.multocidaが分離されるが、同一供試牛から分離された同一菌種の細菌であっても、薬剤感受性が異なる可能性があることから、重症慢性肺炎罹患牛においてSwabによる肺炎原因菌の推定には慎重を要すると思われた。(著者抄録)

Publishing type：Research paper (scientific journal)   International / domestic magazine：Domestic journal  

The purpose of this study was to determine the effect of beta-carotene supplementation to Japanese Black calves on the peripheral blood leukocyte population. Twenty-two Japanese Black calves were alternately assigned to two groups. Eleven calves received 20 mg/d of beta-carotene orally from 2 to 8 wk of age (BC group), and the other 11 calves did not receive (control group). The serum beta-carotene concentration in the BC group at 4, 8 and 12 wk of age were significantly higher than those in the control group (p<0.05). The numbers of CD4+ cells in the BC group were significantly higher than those in the control group at 4 wk of age (p<0.05). These results confirmed that beta-carotene supplementation to Japanese Black calves affected the peripheral blood leukocyte population.

DOI： 10.3177/jnsv.66.381

PubMed

Publishing type：Research paper (scientific journal)   International / domestic magazine：Domestic journal  

The purpose of this study was to clarify the distribution of marbofloxacin (MBFX) within the bronchoalveolar region of calves. Four clinically healthy calves were intramuscularly injected with a single dose of MBFX (2 mg/kg). Samples of plasma and bronchoalveolar lavage fluid (BALF) were obtained for each calf at 0 (before administration), 1, 2, 6 and 24 hr after injection of MBFX. The injections and series of sample collections were conducted and repeated again after two weeks. The results show that the MBFX concentrations in the pulmonary epithelial lining fluid (ELF) were significantly higher than that in plasma and in alveolar cells at 2 hr after injection (P<0.05). For concentrations of MBFX within the ELF, the mean area under the MBFX concentration curve calculated during the 0 to 24 hr timeframe (AUC0-24) was significantly higher than the mean determined from samples collected from the plasma (P<0.05). These results suggest that intramuscularly injected MBFX was well distributed in the bronchoalveolar region.

DOI： 10.1292/jvms.18-0761

PubMed