Isolation of Nicoletella semolina from Equine Tracheal Washes

The aim of the present study was to describe the prevalence of Nicoletella semolina in equine airways and its relationships with cytological evaluation of tracheal wash (TW). Samples were collected in the framework of routine bacteriological diagnostics of equine TW between May 2010 and June 2011. N semolina has been isolated, along with either common pathogens or contaminants, from 19 (1.8%) of the 1,054 TW samples. Median TW neutrophil counts (87.0%) in specimens from N semolina-positive horses were significantly different from those from N semolina-“negative” horses (52.0%). The data presented in this report may suggest considering such bacteria in horses clinically suffering from airway inflammation.     

In-Laboratory Diffraction-Enhanced X-Ray Imaging of an Equine Hoof

We describe and demonstrate the first application of a laboratory-based diffraction-enhanced X-ray imaging instrument for noninvasive equine imaging. A formalin-preserved disarticulated forelimb from a near-term aborted miniature horse fetus was imaged with diffraction-enhanced X-ray imaging. The resultant calculated images—absorption, extinction, refraction, and scattering—are presented, and soft-tissues such as the dorsal digital extensor tendon, articular cartilage, as well as various joint, tendon sheath, and bursa recesses are observed in simultaneous registration with the adjacent dense bone tissue. Radiation dose calculations were performed and a calculated surface dose of 0.6 mGy for the soft muscular tissue was determined for the imaging experiment.     

Comparison of Two Methods for Presurgical Disinfection of the Equine Hoof

OBJECTIVES: To determine for equine hooves the normal resident aerobic bacterial population and the efficacy of 2 methods of disinfection. Study Design-Measurement of total bacterial, gram-positive bacterial, and gram-negative bacterial surface populations from the frog, sole, and hoof wall after each step of 2 different preoperative surgical disinfection techniques. ANIMALS: Six adult horses. METHODS: Hoof wall, sole, and frog samples were collected for quantitative bacteriology before, during, and after 2 multistep antiseptic preparation techniques: Method A-6-minute scrub with povidone-iodine soap, followed by 24-hour submersion in povidone-iodine solution-soaked cotton; and Method B-initial removal of superficial layer of hoof capsule before completing Method A disinfection procedures. RESULTS: Removal of the superficial hoof layer, application of the povidone iodine scrub, and completion of the povidone-iodine soak all significantly (P < .0008) decreased total bacterial numbers. Method B had significantly lower bacterial counts than method A at each consecutive step. Final total bacterial counts remained greater than 10(5) bacteria per gram of tissue regardless of preparation method. CONCLUSIONS: The hoof surface hosts a broad spectrum of aerobic gram-positive and -negative bacteria, many of which are potential pathogens. Bacterial numbers can be significantly reduced by removal of the superficial hoof surface, by application of a povidone-iodine scrub, and by use of a 24-hour povidone-iodine soak. However, bacterial populations >10(5) g per tissue persist after these disinfection procedures. CLINICAL RELEVANCE: Regardless of the preparation methods used in this study, bacterial populations capable of inducing wound infection remain on the hoof capsule.     

大鼠表皮干细胞的分离培养

为探索一种简单而又高效的分离大鼠表皮干细胞(ESCs)的方法,采用酶消化法和组织块法对大鼠表皮干细胞进行分离、培养;并用Ⅳ型胶原快速黏附法筛选大鼠表皮干细胞,免疫组织化学方法检测CK15、P63、β1-integrin和α6-integrin表达情况;同时,测定ESCs单细胞克隆与克隆形成率及表皮干细胞生长曲线.结果表明,采用组织块法和酶消化法均能分离得到大鼠表皮干细胞,获得的细胞生长良好,具有较高的克隆形成率,前者可传至第6代,而后者传至3代～4代便分化;干细胞标记物CK15、P63、β2-integrin和α6-integrin均呈阳性;酶消化法和组织块法均能成功地分离、纯化和培养大鼠表皮干细胞,组织块法相对较好.     

大鼠表皮干细胞的分离及鉴定

在试验中采用组织块法分离培养大鼠表皮干细胞,观察所培养的细胞形态,免疫组织化学方法检测CK15、P63、α6-integrin表达情况并用α6-integrin标记表皮干细胞经流式细胞仪检测其标记率,同时对1~6代表皮干细胞在不同培养时间计数。结果表明,采用组织块法能分离得到大鼠表皮干细胞,且获得的细胞生长良好;干细胞标记物CK15、α6-integrin、P63呈阳性。经流式细胞仪检测得知,黏附细胞被α6-integrin标记的几率为80%,而未黏附细胞被α6-integrin标记的几率仅为2%,二者相差较大。     

In Vitro Fermentative Capacity of Equine Fecal Inocula of 9 fibrous Forages in the Presence of Different Doses of Saccharomyces cerevisiae

This experiment was conducted to evaluate in vitro effects of equine fecal inocula fermentative capacity on 9 fibrous forages in the presence of Saccharomyces cerevisiae. The fibrous feeds were corn stover (Zea mays), oat straw (Avena sativa), sugarcane bagasse and leaves (Saccharum officinarum), llanero grass leaves (Andropogon gayanus), Taiwan grass leaves (Pennisetum purpureum), sorghum straw (Sorghum vulgare), and steria grass leaves (Cynodon plectostachyus). Fibrous feed samples were incubated with several doses of S. cerevisiae; 0 (control), 1.25 (low), 2.5 (medium) and 5 (high) mg/g dry matter (DM) of a commercial yeast product containing 1 × 10¹⁰/g. Fecal inoculum was collected from 4 adult horses were fed on an amount of commercial concentrate and oat hay ad libitum. Gas production (GP) was recorded at 2, 4, 6, 8, 10, 12, 24, and 48 hours post inoculation. An interaction occurred between feeds and yeast dose for fecal pH (P < .01), asymptotic GP (b, ml/g DM); rate of GP (c, /hr); initial delay before GP began (L, hours), GP at 4 hours and 48 hours (P < .01), and GP at 8 hours (P < .01) and at 24 hours (P < .01). Differences in fecal fermentation capacity between the tropical and template grass (P < .05) occurred for fecal pH, c, and GP during first 12 hours, whereas differences occurred (P < .05) between the agriculture byproducts and the grasses for fecal pH, b, and GP from 8 to 48 hours. Fermentation capacity between straws versus not straws (P < .05) differed for fecal pH, b, and GP after 12 hours between straws versus not straws. Addition of S. cerevisiae to Z. mays stover reduced (P < .01) fecal pH and the c fraction with a higher (P < .01) b fraction versus the other feeds. From 4 to 24 hours, S. officinarum bagasse improved GP to the highest values versusS. officinarum leaves. After 24 hours, Z. mays stover had the highest GP, whereas C. plectostachyus leaves had the lowest. There were no differences among the yeast doses for all measured parameters with the exception of L values (linear effect; P < .01). The Z. mays stover had the highest nutritive compared to the other fibrous feeds. However, addition of S. cerevisiae at 2.5 to 5.0 g/kg DM improved fecal fermentation capacity of low-quality forages.     

Comparison of in vivo and in vitro survival and fecundity rates of the poultry red mite, Dermanyssus gallinae

To assist in the testing of possible antigens in developing a vaccine against the poultry red mite (Dermanyssus gallinae De Geer), a rapid and reliable in vitro screening method is critical. This short paper describes how D. gallinae survival and fecundity rates in an in vivo feeding device compared to that of mites fed using an in vitro method. Results showed that survival of fed D. gallinae females and mites overall was greater in vitro, although there was no difference between male survival and fecundity between in vivo and in vitro designs. The in vitro feeding device described therefore has the potential to provide reliable results, comparable to those obtained by in vivo testing, to allow for the rapid screening of D. gallinae antigens.     

马脂肪间充质干细胞的分离培养与生物学特性试验

为马的关节炎、肌腱和韧带损伤的临床治疗提供种子细胞,本试验通过采集马颈部脂肪组织,采用Ⅰ型胶原酶消化法分离脂肪间充质干细胞,并进行传代培养;绘制细胞生长曲线、测定细胞群体倍增时间;通过流式细胞术检测P3代细胞表面标记物,RT-PCR法扩增细胞表面标记物目的基因片段;油红O和茜素红染色法测定脂肪间充质干细胞的成脂和成骨诱导分化能力。结果发现:马脂肪间充质干细胞体外培养条件下呈长梭形和典型的旋涡状,生长状态良好、折光性强;经测定细胞生长曲线呈典型的S型,符合Logistic生长曲线规律;P3、P6和P9代细胞群体倍增时间分别为21.5 h、26 h、36 h;流式细胞术检测结果显示,P3代细胞高表达间充质干细胞表面标志物CD44、CD90和CD105,不表达造血系细胞表面标志物CD45;PCR扩增得到CD44、CD90、CD105和CD73特异性目的片段;油红O和茜素红染色证明,马脂肪间充质干细胞具有成脂和成骨诱导分化能力。     

Evaluation of the in vitro growth-inhibitory effect of epoxomicin on Babesia parasites

Epoxomicin potently and irreversibly inhibits the catalytic activity of proteasomal subunits. Treatment of proliferating cells with epoxomicin results in cell death through accumulation of ubiquinated proteins. Thus, epoxomicin has been proposed as a potential anti-cancer drug. In the present study, the inhibitory effects of epoxomicin on the in vitro growth of bovine and equine Babesia parasites were evaluated. The inhibitory effect of epoxomicin on the in vivo growth of Babesia microti was also assessed. The in vitro growth of five Babesia species that were tested was significantly inhibited (P < 0.05) by nanomolar concentrations of epoxomicin (IC₅₀ values = 21.4 ± 0.2, 4 ± 0.1, 39.5 ± 0.1, 9.7 ± 0.3, and 21.1 ± 0.1 nM for Babesia bovis, Babesia bigemina, Babesia ovata, Babesia caballi, and Babesia equi, respectively). Epoxomicin IC₅₀ values for Babesia parasites were low when compared with diminazene aceturate and tetracycline hydrochloride. Combinations of epoxomicin with diminazene aceturate synergistically potentiated its inhibitory effects in vitro on B. bovis, B. bigemina, and B. caballi. In B. microti-infected mice, epoxomicin caused significant (P < 0.05) inhibition of the growth of B. microti at the non-toxic doses of 0.05 and 0.5 mg/kg BW relative to control groups. Therefore, epoxomicin might be used for treatment of babesiosis.     

西门塔尔牛表皮干细胞的分离培养与生物学特性研究

旨在建立西门塔尔牛表皮干细胞(epidermal stem cell, EpSCs)分离培养体系,探究其生物学特性,为西门塔尔牛种质资源保存提供新方法,为干细胞治疗研究提供种子细胞。采用3～4月龄西门塔尔牛背部表皮,通过酶消化法和组织贴壁法两种方法分离培养西门塔尔牛EpSCs,绘制EpSCs生长曲线探讨其增殖能力。通过免疫荧光鉴定EpSCs表面标记物(ITG β1、P63和CD71),通过RT-PCR分析EpSCs特异性基因(ITG β1、KRT19和ITG α6)表达情况。通过体外诱导EpSCs分化为脂肪细胞、成骨细胞和软骨细胞检测其分化潜能。结果显示,组织块贴壁法获得的EpSCs纯度较低,酶消法获得的EpSCs纯度较高,细胞传代至P28代时开始逐渐老化。EpSCs细胞生长曲线呈“S”型。免疫荧光结果显示EpSCs表面特异性标记物ITG β1、P63和CD71呈阳性表达,RT-PCR结果显示EpSCs高度表达ITG β1、KRT19和ITG α6,不表达CD31基因。EpSCs诱导分化脂肪细胞时,产生可被油红O着色的脂滴,经鉴定细胞阳性表达脂肪细胞PPAR-γ和LPL特异性基因。Ep...     

蹄变形奶牛血液流变学的研究

为了更深入探讨奶牛变形蹄的发病机理及对奶牛变形蹄的预防和治疗提供更科学的理论依据。选择长沙地区20头中国荷斯坦牛，系统检测了变形蹄和正常蹄奶牛在夏季（6月份）和冬季（12月份）血液流变学主要指标。夏季时，1S-1、3S-1、5S-1、10S-1、30S-1、60S-1切变率下，变形蹄奶牛全血粘度高于正常蹄奶牛，有显著差异（P〈0．05）。变形蹄奶牛低切全血还原粘度极显著高于正常蹄奶牛（P〈0．01）。变形蹄奶牛红细胞压积高于正常蹄奶牛，差异显著（P〈0．05）。冬季时，5S^-1、10S^-1、30S^-1、200S^-1切变率下，变形蹄奶牛全血粘度高于正常蹄奶牛（P〈0．05）。变形蹄奶牛血浆粘度高于正常蹄奶牛，有显著差异（P〈0．05）。变形蹄奶牛红细胞沉降率较快，也有显著差异（P〈0．05）。变形蹄奶牛与全血粘度增高和红细胞聚集性增强具有相关性。     

Methods for Equine Preantral Follicles Isolation: Quantitative Aspects

The aim of this study was to test the use of mechanical and mechanical‐enzymatic methods, saline solution (SS), and PBS solution for the manipulation and isolation of mare ovarian preantral follicles (PAFs). The ovaries were subjected to mechanical isolation (mixer) alone or in association with enzymatic digestion (collagenase). Incubation times of 10 and 20 min were employed. In the first group, 4.1 ± 4.9 PAFs were harvested with the mechanical‐enzymatic method vs 71.1 ± 19.2 with the mechanical procedure, showing a significant difference between methods; using SS and PBS, these numbers were 35.7 ± 34.3 and 39.6 ± 39.6, respectively, with no significant difference between solutions. In the second group, there was significant difference between methods, with 7.1 ± 10.6 follicles harvested with the mechanical‐enzymatic method vs 63.2 ± 22.9 with the mechanical procedure; using SS and PBS, means were 35.5 ± 36.4 and 34.9 ± 31.1, respectively. The mechanical method proved more effective than the mechanical‐enzymatic approach. Both SS and PBS can be used as a media for equine PAFs preparation.