Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

The relationship between the size of caudolateral curvilinear osteophyte of the canine femoral neck and the radiographic view

Caudolateral curvilinear osteophyte (CCO), an osteophyte at the site of joint capsule attachment on the caudal aspect of the femoral neck, has been advocated as a radiographic criterion for coxofemoral subluxation. The correlation between the presence of CCO on radiographs (radiographic-CCO), the size of the CCO (CCO index) on three-dimensional computed tomographic (CT) images, and hip evaluation using transverse CT images was assessed in 22 Border Collies. CCOs were detected on the radiographs and CT images of 32% and 100% femurs, respectively. The CCO index correlated significantly with radiographic-CCO, but a large CCO index did not necessarily imply that the CCO was visible on radiographs. Hence, radiographic-CCO findings should be used cautiously in hip evaluation of Border Collies.     

In vitro effects of plant and mushroom extracts on immunological function of chicken lymphocytes and macrophages

1.?The present study was conducted to examine the effects of organic extracts from milk thistle (Silybum marianum), turmeric (Curcuma longa), reishi mushroom (Ganoderma lucidum), and shiitake mushroom (Lentinus edodes) on innate immunity and tumor cell viability.

2.?Innate immunity was measured by lymphocyte proliferation and nitric oxide production by macrophages, and the inhibitory effect on tumor cell growth was assessed using a non-radioactive assay. For measuring the cytokine levels in the HD11 macrophages which were treated with extracts of turmeric or shiitake mushroom, the levels of mRNAs for interferon-α (IFN- α), interleukin-1β (IL-1β), IL-6, IL-12, IL-15, IL-18, and tumor necrosis factor superfamily 15 (TNFSF15) were quantified by real time RT-PCR.

3.?In vitro culture of chicken spleen lymphocytes with extracts of milk thistle, turmeric, and shiitake and reishi mushrooms induced significantly higher cell proliferation compared with the untreated control cells. Stimulation of macrophages with extracts of milk thistle and shiitake and reishi mushrooms, but not turmeric, resulted in robust nitric oxide production to levels that were similar with those induced by recombinant chicken interferon-γ. All extracts uniformly inhibited the growth of chicken tumor cells in vitro at the concentration of 6·3 through 100 µg/ml. Finally, the levels of mRNAs encoding IL-1β, IL-6, IL-12, IL-18, and TNFSF15 were enhanced in macrophages that were treated with extracts of turmeric or shiitake mushroom compared with the untreated control.

4.?These results document the immunologically-based enhancement of innate immunity in chickens by extracts of plants and mushrooms with known medicinal properties in vitro. In vivo studies are being planned to delineate the cellular and molecular mechanisms responsible for their mechanism of action.     

茶多酚对高酒糟日粮肉鸡生产性能和抗氧化特性的影响

本研究旨在评估添加茶多酚对高酒糟(distillers dried grain with solubles DDGS)日粮肉鸡生产性能和抗氧化特性的影响。将400只21日龄肉鸡随机分为5个处理,每个处理4个重复,每个重复20只。正对照组为玉米-豆粕型日粮,负对照组为玉米-豆粕-DDGS型日粮(15%DDGS),处理组分别在负对照日粮中添加100、300、500 mg/kg茶多酚。结果表明,与正对照组相比,后期日粮中添加15%DDGS对肉鸡生产性能无显著影响(P>0.05),但血液的GSH-Px降低8.31%(P<0.01),胸腿肉T-AOC含量降低,腿肉MDA值显著升高(P<0.05)。添加茶多酚提高了血液的T-AOC和GSH-Px活性(P<0.05)。随着茶多酚添加剂量的升高,T-AOC和GSH-Px有升高的趋势,但各水平组之间差异不显著;茶多酚添加300 mg/kg以上能显著降低胸腿肌MDA值(P<0.05),提高T-AOC含量,改善肉质的抗氧化特性。本研究结果表明,在肉鸡高DDGS日粮中添加茶多酚可以改善肉鸡机体及肉质的抗氧化特性。     

Distribution and characterization of class 1 integrons in Salmonella enterica serotype Gallinarum biotype Gallinarum

Fowl typhoid caused by Salmonella enterica subsp. enterica serotype Gallinarum biotype Gallinarum is the most important chicken disease in Korea. Due to appearance of new or multiple antibiotics resistances in the recently isolated strains, it was difficult to control the disease using antibiotics in our country. Therefore, the prevalence and genetic contents of class 1 integrons in biotype Gallinarum isolated between 1992 and 2001 were investigated by PCR and direct sequencing, respectively. Out of 90 strains, 35 (39%) carried class 1 integrons. The 1.0, 1.6 and 2.0kbp amplicons were amplified in 32 strains (36%), 2 strains (2%) and 1 strain (1%), respectively. The 1.0, 1.6 and 2.0 kbp amplicons contained one (aadA1a), two (aadB-aadA1b) and three cassettes (dhfrXII-orfF-aadA2), respectively, providing resistances against aminoglycosides (aadA1a, aadA1b, aadB, and aadA2) and trimethoprim (dhfrXII). The integron-carrying strains of biotype Gallinarum appeared in 1996 and acquired additional cassettes in 2000. Although the resistances to ampicillin, tetracycline and chloramphenicol are unrelated to class 1 integrons, relatively high prevalence of integron in biotype Gallinarum may be a dormant threat to the chemotherapy of the disease in the near future because of potency to acquire additional antibiotics resistances.     

马立克氏病病毒致瘤基因meq功能研究的概述

马立克氏病病毒 (Ｍａｒｅｋ’ｓＤｉｓｅａｓｅＶｉｒｕｓ,ＭＤＶ)之众多已得到鉴定的基因中 ,目前有 3个基因即 132 ｂｐｒ、ｐｐ38、ｍｅｑ被认为可能与致瘤相关。其中 ,ｍｅｑ与 132 ｂｐｒ两个基因是ＭＤＶ 1所特有 ,已证实 132 ｂｐｒ的拷贝数与毒株的致弱程度有关 ,ｐｐ38则被证实可抑制机体的免疫应答。ｍｅｑ基因由于具有与致瘤基因ｊｕｎ/ ｆｏｓ家族类似的分子结构 ,因此 ,我们有理由认为ｍｅｑ基因在ＭＤＶ致病、致瘤中可能发挥重要的作用。本课题在国家自然科学基金和广西自然科学基金的资助下 ,历经 5年由三个单位共同攻关…     

Cutaneous lymphoma in a juvenile dog

Abstract: An 18-month-old male Doberman Pinscher was referred to the Veterinary Medical Teaching Hospital of the College of Veterinary Medicine for an erythemic nodular mass on the right forelimb. The mass was diagnosed as cutaneous lymphoma, based on cytologic examination of a mass aspirate and histopathology. Using immunohistochemistry the neoplastic cells were positive for CD3 but negative for CD79a, E-cadherin, and pancytokeratin, confirming their origin as T lymphocytes. No tumor recurrence was noted 18 months after surgery. To our knowledge, this is the first report of a solitary nodular form of cutaneous lymphoma in a young dog.     

Genotypic prevalence of the adhesin involved in diffuse adherence in Escherichia coli isolates in pre-weaned pigs with diarrhoea in Korea

A total of 1002 Escherichia coli strains isolated from pre-weaned pigs with diarrhoea on 1114 swine farms were screened for the presence of the adhesin involved in diffuse adherence (AIDA) gene by polymerase chain reaction (PCR). Escherichia coli isolates that carried AIDA genes were also tested by PCR for the detection of five fimbriae (F4, F5, F6, F18 and F41), heat-stable (STa, STb) and heat-labile (LT) enterotoxin, enteroaggregative E. coli heat-stable enterotoxin 1 (EAST1), and Shiga toxin 2 oedema disease (Stx2e) genes. Twenty-three (2.3%) of the 1002 E. coli isolates carried the gene for AIDA. Among 23 isolates shown to carry genes for AIDA, three carried the AIDA gene as the only shown virulence factor. Other isolates carried other virulence factor genes in addition to AIDA. Four isolates carried genes for at least one of the fimbrial adhesins and enterotoxins. Sixteen isolates carried genes for enterotoxins only. The AIDA may represent an additional virulence determinant in pre-weaned pigs with diarrhoea.