Integrated standardization concept for Angelica botanicals using quantitative NMR

Despite numerous in vitro/vivo and phytochemical studies, the active constituents of Angelica sinensis (AS) have not been conclusively identified for the standardization to bioactive markers. Phytochemical analyses of AS extracts and fractions that demonstrate activity in a panel of in vitro bioassays, have repeatedly pointed to ligustilide as being (associated with) the active principle(s). Due to the chemical instability of ligustilide and related issues in GC/LC analyses, new methods capable of quantifying ligustilide in mixtures that do not rely on an identical reference standard are in high demand. This study demonstrates how NMR can satisfy the requirement for simultaneous, multi-target quantification and qualitative identification. First, the AS activity was concentrated into a single fraction by RP-solid-phase extraction, as confirmed by an alkaline phosphatase, (anti-)estrogenicity and cytotoxicity assay. Next, a quantitative ¹H NMR (qHNMR) method was established and validated using standard compounds and comparing processing methods. Subsequent 1D/2D NMR and qHNMR analysis led to the identification and quantification of ligustilide and other minor components in the active fraction, and to the development of quality criteria for authentic AS preparations. The absolute and relative quantities of ligustilide, six minor alkyl phthalides, and groups of phenylpropanoids, polyynes, and poly-unsaturated fatty acids were measured by a combination of qHNMR and 2D COSY. The qNMR approach enables multi-target quality control of the bioactive fraction, and enables the integrated biological and chemical standardization of AS botanicals. This methodology can potentially be transferred to other botanicals with active principles that act synergistically, or that contain closely related and/or constituents, which have not been conclusively identified as the active principles.     

Limited susceptibility of three different mouse (Mus musculus) lines to Porcine circovirus-2 infection and associated lesions

Porcine circovirus associated disease (PCVAD), a major global problem for pork producers, is characterized microscopically by depletion and histiocytic replacement of follicles in the lymphoid tissues. The objectives of this study were to determine 1) if Porcine circovirus-2 (PCV-2) inoculated mice (Mus musculus) can develop PCV-2 associated lymphoid lesions and serve as a model for PCVAD, and 2) if differences in PCV-2 host susceptibility exist among mice lines. Three groups (n = 48/group) of 4-wk-old male mice were used: BALB/c, C57BL/6, and C3H/HeJ. A 2 × 2 factorial analysis was designed for each group using PCV-2 inoculation and keyhole limpet hemocyanin in incomplete Freund’s adjuvant injections on day 0 and 7 as factors. Necropsies were performed on days 12, 17, 22, 27, 32, and 37. Serum samples collected at each necropsy tested negative for anti-IgG PCV-2 antibodies in all mice at all time points by 2 different PCV-2 enzyme-linked immunosorbent assays (ELISA). The PCV-2 DNA was detected by polymerase chain reaction (PCR) in 93% (100/108) of tissues and 42.6% (46/108) of serum samples from PCV-2-inoculated mice from days 12 to 37. Microscopic lesions consistent with PCV-2 infection were not observed in any mice and PCV-2 DNA and PCV-2 antigen were not detected in tissues by in-situ-hybridization or immunohistochemistry assays, respectively. Based on incidence of PCV-2 DNA in serum samples, the C57BL/6 mouse line was more resistant to PCV-2 infection than the other lines. The results indicate the mouse model likely has limited utility to advance understanding of the pathogenesis of PCV-2 associated lesions, but mice could potentially be important in the epidemiology of PCV-2.     

Infectivity of porcine circovirus type 2 DNA in semen from experimentally-infected boars

Porcine circovirus type 2 (PCV2) is an economically important pathogen. It has been demonstrated that PCV2 DNA can be detected in boar semen by PCR; however, the biological relevance of this is unknown. The objectives of this study were to determine if semen positive for PCV2 DNA is infectious (1) in a swine bioassay, or (2) when used for artificial insemination. For the first objective, 4-week-old pigs were inoculated intraperitoneally with PCV2 DNA-negative (bioassay-control; n = 3), PCV2a DNA-positive (bioassay-PCV2a; n = 3), or PCV2b DNA-positive (bioassay-PCV2b; n = 3) raw semen, or PCV2 live virus (bioassay-positive; n = 3), respectively. Pigs inoculated with PCV2 DNA-positive semen and PCV2 live virus became viremic and developed anti-PCV2 antibodies indicating that the PCV2 DNA present in semen was infectious. For the second objective, three Landrace gilts were inseminated with PCV2 DNA-negative semen (gilts-controls) from experimentally-infected boars, and six gilts were artificially inseminated with semen positive for PCV2a DNA (gilts-PCV2a; n = 3) or PCV2b DNA (gilts-PCV2b; n = 3). Serum samples collected from the gilts in all groups remained negative for anti-PCV2 antibodies for the duration of the experiment. In addition, fetal serum samples from all 105-day-gestation fetuses were negative for anti-PCV2 antibodies or PCV2 DNA. Under the conditions of this study, PCV2 DNA-positive semen was not infectious when used to artificially inseminate gilts; however, it was demonstrated to be infectious in a swine bioassay model and therefore is a potential means of PCV2 transmission amongst swine herds.     

Prevalence and radiologic and histologic appearance of vesicourachal diverticula in dogs without clinical signs of urinary tract disease

OBJECTIVE: To determine prevalence and radiologic and histologic appearance of vesicourachal diverticula in dogs without clinical signs of urinary tract disease. DESIGN: Original study. ANIMALS: 50 dogs between 4 months and 17 years old representing 22 breeds that had been euthanatized for unrelated reasons; none of the dogs had a history or clinical signs of urinary tract disease. PROCEDURE: Retrograde positive-contrast radiography was performed, and radiographs were examined for macroscopic diverticula. Necropsy specimens from the urinary bladder vertex were examined by means of light microscopy for diverticula and signs of inflammation. RESULTS: 17 of the 50 (34%) dogs had vesicourachal diverticula, and 1 additional dog had a urachal cyst. Fifteen of the 17 diverticula were macroscopic; surface area of the diverticulum could be measured radiographically in 13 of these dogs and ranged from 1 to 90 mm2. The remaining 2 diverticula were microscopic. Sixteen diverticula were intramural and 1 was extramural. Light microscopic signs of bladder wall inflammation could be detected in 5 dogs, 4 of which had macroscopic diverticula. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a high percentage of dogs without clinical signs of urinary tract disease may have vesicourachal [corrected] diverticula. Further studies are needed to determine the clinical relevance of vesicourethral diverticula in dogs.     

Protein-losing enteropathy caused by Lawsonia intracellularis in a weanling foal

A 5-month-old Morgan filly was presented to the Atlantic Veterinary College with a history of lethargy, fever, depression, anorexia, and dependent ventral edema. Diagnostic tests revealed severe inflammation, hypoproteinemia, and thickened small intestinal loops. Protein-losing enteropathy caused by Lawsonia intracellularis was diagnosed and treated successfully with erythromycin-rifampin.     

Aqueous misdirection and ciliary block (malignant) glaucoma after cataract removal in a llama

Ciliary block (malignant) glaucoma is caused by the posterior flow of aqueous humor into or behind the vitreous. It is a rare complication of anterior segment surgery in humans. This is a report of ciliary block glaucoma occurring as a postoperative complication of cataract surgery in a llama. Medical management was ineffective in lowering intraocular pressure (IOP). Posterior capsulotomy and anterior hyaloid disruption with a 22-gauge needle, in addition to medical management maintained IOP in the normal range until anterior vitrectomy could be performed. After vitrectomy, glaucoma resolved and medical treatment was no longer necessary.     

猪圆环病毒病及其相关疾病

猪Ⅱ型圆环病毒(PCV-2)仅靠其自身的力量极少可引起健康猪出现临床疾病。本文主要介绍PCV-2的协同感染因子,因为这些或/和其它环境因子对诱发与PCV-2有关的完整临床症状是非常重要的。     

Prevalence of Campylobacter spp. and Yersinia spp. in the pig production

The aim of this study was to determine the prevalence of Campylobacter spp. and Yersinia spp. in a total of 1,040 faecal samples taken from animals at different ages from four farrowing and twelve fattening herds. In the farrowing unit, faeces were collected from 68 sows (faecal samples) and 256 suckling piglets (rectal swab samples). Further samples were collected from 362 growing and 354 finishing pigs (rectal swab samples). Additionally, 56 feed and environmental samples were collected. During the slaughtering process, 122 pigs and their carcasses respectively, were sampled three times. Finally, 86 meat and minced meat samples were taken from 34 retail stores. Campylobacter spp. were isolated in sows (33.8 %), piglets (80.9 %), growing (89.2 %) and finishing (64.7 %) pigs. Yersinia spp. were detected in growing (15.2 %) and finishing (13.3 %) pigs only. After twelve hours of chilling neither Campylobacter spp. nor Yersinia spp. were detected. In raw meat samples, Campylobacter spp. were isolated from one liver sample and Yersinia enterocolitica from two meat samples. Common slaughter techniques and hygiene procedures may be effective tools to reduce the risk of contamination and recontamination of meat products since Campylobacter spp. and Yersinia spp. were found only sporadically in raw meat samples.     

Morphological and molecular identification of Eutypa lata on grapevine in Serbia

Journal of Plant Diseases and Protection - During a survey from 2004 to 2016, symptoms of grapevine dieback were observed in six vineyards in Serbia. Symptoms initially appeared as small,...     

火鸡球虫病的防制

给火鸡造成重大经济损失的球虫病极其普遍 ,但往往难以诊断。尤其当该病对火鸡日增重和死亡率影响不明显时 ,对混饲抗球虫药的防制效果则难以判定。因此 ,一旦怀疑球虫病发生时 ,就广泛采用常规混饲抗球虫药的方法进行“诊断性治疗”。一般 3～ 5周龄左右可能是火鸡球虫病的第1个高发期。但最近有报道表明 ,7周龄左右也可发病 ,但早期暴发过球虫病的鸡有可能产生抵抗力。在德国进行的 1次球虫病试验中 ,把 1群健康火鸡放入 1栋曾严重感染过球虫病的火鸡舍 ,为确保自然感染 ,此鸡舍仅采取了简单消毒措施 38d后 ,将这批火鸡从试验场地被移到铺…