Partial purification and characterization of chicken interleukin-2.

Chicken interleukin 2 (IL-2) activity was partially purified from conditioned medium produced by culturing chicken splenic lymphocytes in the presence of concanavalin A. The purification procedure included sequential steps of gel filtration chromatography, reverse-phase high-pressure liquid chromatography, and phenyl-sepharose chromatography. Two peaks of IL-2 activity with apparent mol. wt. ranges of 36-39 kD and 17.5-25 kD were eluted from the Sephadex G100 gel filtration column. An increase in IL-2 spec. act. from 14 U mg-1 to between 2000 and 20,000 U mg-1 was obtained for the Sephadex G100 column peaks when subjected to the subsequent steps of the purification procedure. Alkylative reduction of the higher mol. wt. Sephadex G100 column peak (followed by re-chromatography with Sephadex G100), resulted in generation of the lower (17.5 kD) mol. wt. peak, indicating that chicken IL-2 is capable of either dimerizing or forming aggregates with other proteins. Elution of the lower mol. wt. IL-2 activity from a non-reducing sodium dodecyl sulfate-polyacrylamide gel demonstrated an apparent mol. wt. for chicken IL-2 of 20 kD, which confirmed the range of 17.5-25 kD seen with gel filtration.     

圈养红斑羚冬季活动节律及活动时间分配

2010年冬季,采用瞬时扫描取样法对上海动物园繁殖场内5只红斑羚进行活动节律和活动时间分配的研究。观察结果显示:冬季红斑羚有54.17%的时间趴卧休息,其次是摄食行为(22.17%)和站立行为(15.34%),运动行为、修饰行为和其他行为占的比例非常小,分别为5.32%、2.71%和0.29%。卧息行为一天中有2个高峰,分别是7:00之前和摄食之后10:00~13:00时之间,持续时间较长。摄食行为也有2个高峰,分别是9:00~10:00时和14:00时以后。站立行为和运动行为伴随着摄食行为,同样在8:00~9:00时和14:00~15:00时形成了2个高峰,只是运动行为所占比例很小,高峰期不明显。修饰行为和其他行为是随机发生的,分布比较平均。     

三种绦虫蛋白质等电聚焦电泳比较分析

应用PAGE-IEF分析了三种绦虫的可溶性蛋白质。猪囊尾蚴头节囊壁、囊液,细颈囊尾蚴头节囊壁、囊液和曼氏迭宫绦虫成虫蛋白质区带数分别为30、25、29、18和24～29条,主带(峰)数分别为10、9、9、7和12个,面积最大峰分别为J、B、H、F和E峰。五种样品电泳谱在总带数、各pH区带数、主带(峰)数和分布、面积最大峰都明显不同,可供鉴定和鉴别。细颈囊尾蚴蛋白质电泳分析与猪囊尾蚴的比较分析,以及曼氏迭宫绦虫蛋白质电泳分析系国内外首次报道。     

Separation of parasite antigens by molecular exclusion, anion exchange, and chromatofocusing utilizing FPLC protein fractionation systems

Excretory-secretory products (ESP) were harvested from balanced salt solutions in which adult Fasciola hepatica had been incubated for 4-6 h at 37 degrees C. The ESP was fractionated by standard low pressure molecular exclusion chromatography and FPLC (fast protein liquid chromatography) using the principles of molecular exclusion, anion exchange, and chromatofocusing. The dot-enzyme-linked immunosorbent assay (Dot-ELISA) was used to demonstrate the immunoreactivity of eluted fractions. Compared to Sephacryl S-200, separation by Superose-6 (FPLC) was faster and resolved more peaks (four with Sephacryl S-200 and nine with Superose-6). Peaks from Sephacryl S-200 were resolved by the first anion exchange (Mono Q) separation into seven peaks; when these peaks were subjected to a second anion exchange, 15 peaks were resolved. Thirty-eight peaks were resolved by chromatofocusing (Mono P) in the pH range 7-4. Immunoreactive fractions from narrow-range (single pH unit) chromatofocusing were identified by the Dot-ELISA. The FPLC system proved to be a means of rapid and high resolution separation of F. hepatica antigens.     

Some physicochemical properties of the virus of rabbit haemorrhagic disease.

Purified and concentrated preparations of virus from liver extracts of infected rabbits contain virus specific components with sedimentation coefficients of about 175, 110 and sometimes 133S and more slow units. Full and empty virus particles with a diameter of about 34 nm were shown electron microscopically in the corresponding 175 and 110S fractions of the sucrose density gradient. The average of buoyant density of the 175, 133, 110S and more slow units are 1.36, 1.32 and 1.31 g/ml respectively. The extinction coefficient E260 nm is 4.3 +/- 0.7 cm2/mg. The RNA content is 17 +/- 4%. SDS-PAGE shows a "65" kD protein as a single or major component. Beside smaller polypeptides with lower intensities, the 67 kD polypeptide reacts positively in the Western blot with polyclonal antibodies of rabbits. The molecular weight of the virus is 15 +/- 4 x 10(6)D. The pH stability of the 175S unit was also tested.     

Blood acid-base status in fattening pigs in exhausting physical stress of high intensity]

Reported in this paper are changes in the acid-base state and lactic concentration of blood and muscle samples taken from 28 pigs prior to, during, and after high-intensity exhausting stress applied at ambient temperatures between 17 degrees C and 21 degrees C and at 35 degrees C. Close correlations were found to exist between acid-base states, on the one hand, and lactic acid concentrations, on the other. The lactic acid concentrations reached in blood plasma were between 30 and 40 mHol/1, pH being below 7.0 and BE values being smaller than -22 mHol/1. Close relationships were established between the lactic acid concentration of venous blood plasma, blood pH, and the pH of the musculus longissimus dorsi. Respiratory compensation was of minor importance with ambient temperatures between 17 degrees C and 21 degrees C, but it was of considerable effectiveness at 35 degrees C.     

Gonadotrophin concentrations during growth and maturation in domestic turkeys

1. Circulating immuno-reactive-luteinising hormone (LH) and follicle stimulating hormone (FSH) were measured between hatching and either 17 or 30 weeks of age in turkeys. 2. In one experiment both sexes were reared together. The pattern of changes in gonadotrophin concentration with age consisted of high LH concentrations at 3 to 8 and 16 to 19 weeks old and FSH peaks between weeks 3 and 10 and at weeks 18 and 19. 3. In two separate experiments male and female birds were reared separately for 17 weeks. In these turkeys the LH concentration was high for 3 weeks following hatching and also after 10 weeks, while FSH was elevated at weeks 10 and 15 in females and at weeks 7 and 13 in males. 4. There were lower plasma LH concentrations in turkeys which had received testosterone implants while the FSH concentration was elevated. 5. Pinealectomy in female turkeys led to increased FSH concentrations at 2 weeks but depressed FSH concentrations thereafter.     

家蚕孵化酶样基因BmHEL的原核表达及蛋白纯化与功能鉴定

孵化酶(hatchingenzyme,HE)是后生动物卵孵化过程中起关键作用的一类蛋白酶。在已完成全长967bp的家蚕孵化酶样基因(BmHEL)克隆及其转录特性分析的基础上,研究了该基因在原核表达系统的表达、产物纯化及其对家蚕卵壳的降解功能。以全长BmHEL质粒DNA为模板,用PCR方法分别获得了该基因的开放阅读框(BmHELORF,885bp)、缺失信号肽编码区(BmHELa,834bp)和推测成熟酶功能编码区(BmHELb,624bp)的3个片段,并亚克隆入原核表达载体pET28a(+),在大肠杆菌(E.coli)BL21中进行表达。表达的3种外源蛋白均以包涵体形式存在,获得含有6×His-Tag标签的融合蛋白,其分子质量分别为33.4、31.8、24.0kD。通过Westernblot方法检测到了用Ni-NTA亲和层析柱纯化的目的蛋白。纯化复性后的3种孵化酶对家蚕卵壳底物有一定降解活性,其中BmHELORF的活性最高,BmHELb的活性最低。在pH7.1反应条件下3种酶的活性比pH9.5时高。研究结果在蛋白质水平上进一步证实家蚕孵化酶样蛋白参与了蚕卵孵化这一重要生命过程。     

Comparison and partial characterization of the protein profiles and outer membrane antigens of Actinobacillus species isolated from ram lambs with epididymitis

Three monoclonal antibodies (LG17, LG30, LG33) were used in the indirect fluorescent antibody test, the ELISA, and the immunoelectrotransfer blot technique to identify group-specific and strain-specific epitopes on the outer membranes of Actinobacillus seminis, A actinomycetemcomitans, and 17 field isolates of Actinobacillus spp. The field isolates had been obtained by bacteriologic culture of specimens from ram lambs with epididymitis. Only antibody LG33 consistently had specificity for an outer membrane epitope shared by most of the bacterial isolates tested. Staining of polyacrylamide gels with periodic acid-Schiff reagent, Sudan black B, and Coomassie brilliant blue R250 indicated that target antigens for antibodies LG17 and LG33 contained carbohydrate and lipoprotein components, respectively. The chemical composition of the LG30 target antigen was not determined because of its instability after exposure to sodium dodecyl sulfate. Discontinuous-gradient polyacrylamide gel electrophoresis in sodium dodecyl sulfate and spectrophotometric scans of the gels were used to analyze n-octyl-beta-D-glucopyranoside protein extracts from A seminis, A actinomycetemcomitans, and 13 representative field isolates of Actinobacillus spp. Bacterial isolates could be grouped according to their protein profiles. The first group consisted of A seminis, A actinomycetemcomitans, and 7 field isolates of Actinobacillus spp, all of which shared common protein bands with molecular masses of approximately 94 kilodaltons (kD), 64 kD, 60 kD, 52 kD, 44 kD, and 26 kD. The second group was composed of 6 field isolates, each with unique protein profiles; isolates had relatively few protein bands in common. These data suggested that members of the genus Actinobacillus cultured from ram lambs with epididymitis probably include a number of various strains.     

Concentrations of oestradiol-17 beta in plasma and milk and progesterone in plasma during the oestrus cycle and in early pregnancy in goats

Pre-ovulatory peaks in oestradiol-17 beta concentrations were observed on days 1 or 2 and post-ovulatory peaks between days 4 and 7, both in jugular venous plasma and defatted milk, day 1 being the day of the onset of oestrus in the goats. Mean values of the magnitudes of these concentration peaks and of their timing (relative to oestrus) during the oestrus cycle did not differ significantly (P greater than 0.05) from those when the goats were mated and became pregnant. Pre-ovulatory oestradiol-17 beta peaks were invariably greater than the corresponding post-ovulatory peaks, as were peak concentrations in plasma relative to those in defatted milk collected on the same day. Mean intervals between the pre- and post-ovulatory peaks in oestradiol-17 beta concentrations were respectively 4.2 days for plasma and 4.0 days for defatted milk. Concentrations of oestradiol-17 beta in jugular venous plasma and defatted milk were strongly correlated: rank correlation coefficients for the three goats studied were 0.871, 0.668 and 0.739. It is suggested that in goats, as in cattle, ovarian follicular oestradiol-17 beta secretion approaching pre-ovulatory level is restored by 4 days after oestrus and its rapid decline after this time may be due to the inhibitory influence of the rapidly rising plasma progesterone concentration.     

Eperythrozoon ovis — A blood parasite of sheep

Eleven goats given dog-derived Sarcocystis sporocysts, showed illness from day 18. Doses of 5 × 10⁶ sporocysts caused a progressive temperature rise, with peaks at 6,11 and 18 days. Necropsy at 18 and 19 days revealed multiple petechiation and schizogony in endothelial cells. In goats given smaller doses, Hb, PCV and TP levels fell from day 17. IFAT titres rose from day 28.     

Determination of the estrogenic activity of wild phytoestrogen-rich Pueraria mirifica by MCF-7 proliferation assay

The aim of this study was to evaluate the estrogenic activity of tuberous samples of wild, phytoestrogen-rich Pueraria mirifica collected from 28 out of 76 provinces of Thailand by MCF-7 proliferation assay. The plant extracts were administered to MCF-7, ER alpha positive human mammary adenocarcinoma cell cultures, for 3 days at dosages of 0.1, 1, 10, 100 and 1,000 microg/ml and were compared with 17 beta-estradiol at concentrations of 10(-12)-10(-6) M. The mean P. mirifica population at 1 mug/ml exhibited significant proliferation. Two plant samples exhibited levels of proliferation in MCF-7 that were similar to 17beta-estradiol. The mean P. mirifica populations at 100 and 1,000 microg/ml exhibited significant cytotoxicity in MCF-7. Analysis of the estrogenic activity of puerarin, representative of major isoflavonoids in P. mirifica tubers, revealed proliferation in MCF-7 only at the highest dose (10(-6) M) that was 10(2)-10(5) times less active than 17 beta-estradiol. Puerarin and 17 beta-estradiol at concentration of 10(-12)-10(-6) M exhibited no cytotoxicity in MCF-7.     

圈养麇鹿冬季昼间行为及活动规律

2008年11-12月在济南动物园,采用瞬时扫描取样法、焦点动物取样法和全事件取样法,对9只圈养麋鹿的行为及活动规律进行了研究。结果表明,麇鹿冬季行为时间分配比例依次为卧息（36．45％±7．34％）、取食（24．79％±8．11％）、运动（18．67％±5．93％）、反刍（17．50％±5．52％）、其他（2．59％±0．87％）。在冬季麋鹿的卧息在1d内有1个高峰期（12：30～14：30）,取食在1d中有2个高峰期（8：30～9：30和17：30～18：30）,占糜鹿各种行为比例最多的2种行为在1d中是交替出现的,运动在10：30和15：30各有1个较为明显的高峰期,反刍在1d中有1个高峰期（9：30～11：30）,其他行为在1d中发生的较为随机,没有明显的高峰期。年龄因素对糜鹿冬季昼间的卧息、取食和运动行为的影响差异极显著（P〈0．01）,对其他行为影响显著（P〈0．05）,对反刍行为影响不显著（P〉0．05）;性别因素对糜鹿冬季昼间的行为影响不显著（P〉0．05）。     

鸡西机场苍鹭日活动节律的研究

利用样线法调查了鸡西机场周边苍鹭的分布及数量,对栖息在机场西南穆棱河旁的苍鹭进行了活动规律和行为节律的观察,在2011年10月3～7日,通过定点连续观察法记录苍鹭的日活动节律。研究结果显示,苍鹭飞行行为高峰值出现在6：00～8：30、12：00～13：00和15：30～17：00,苍鹭日常各种行为中休息行为、站立行为和理羽行为所占时间比例最高,为82%,晒翅行为在9：00～10：00达到高峰,为31%,其平均飞行高度为65 m,即对机场航班有一定威胁,应引起有关部门注意。     

Immunohistochemical localization of fibroblast growth factor 18 in hair follicles of healthy beagle dogs

Increasing emphasis is being placed on the role of fibroblast growth factors (FGFs) in hair follicle cycling. In mice, expression of FGF18 mRNA peaks during the late telogen phase, leading to the hypothesis that FGF plays a role in anagen induction. There are no data on the presence of FGF18 in dogs. The main objective of this study was to identify and locate FGF18 in the canine hair follicle. The second objective was to assess potential differences in FGF18 concentration between biopsies taken in winter and summer, shoulder and flank regions, and between different sexes. Skin tissue from 10 healthy beagle dogs (three intact females, three spayed females and four intact males) was collected from the shoulder and flank. The biopsies were collected in February and August on day 0, after which the dogs were clipped and biopsies collected again from the shoulder and flank on days 1, 3, 7 and 17. Paraffin sections (4 μm thick) of the biopsies were stained with an anti-FGF18 antibody. The FGF18-positive cells were counted in the hair follicle epithelium from seven follicular units of each biopsy. Fibroblast growth factor 18 was detected as granular cytoplasmatic staining in follicles at the level of the inner root sheath, and rarely in the outer root sheath and dermal papilla. It was also detected in the apocrine glands, in arrector pili muscles and in vascular endothelial cells. There was no statistical difference in the number of FGF18-positive cells or follicles between sexes, different anatomical locations, seasons or the consecutive days of sampling.     

Effects of blood meal pH and irradiation on nursery pig performance

A total of 720 nursery pigs in three experiments were used to evaluate the effects of blood meal with different pH (a result of predrying storage time) and irradiation of spray-dried blood meal in nursery pig diets. In Exp. 1, 240 barrows and gilts (17 +/- 2 d of age at weaning) were used to determine the effects of blood meal pH (7.4 to 5.9) in diets fed from d 10 to 31 postweaning (7.0 to 16.3 kg of BW). Different lots of dried blood meal were sampled to provide a range in pH. Overall (d 0 to 21), pigs fed diets containing blood meal had greater ADG (P < 0.05) and ADFI (P < 0.05) than pigs fed diets without blood meal. Ammonia concentrations in blood meal rose as pH decreased. However, blood meal pH did not influence (P > 0.16) ADG, ADFI, or gain:feed (G:F). In Exp. 2, 180 barrows (17 +/- 2 d of age at weaning) were used to determine the effects of post drying pH (7.6 to 5.9) and irradiation (gamma ray, 9.5 kGy) of blood meal on growth performance of nursery pigs from d 5 to 19 postweaning (6.8 to 10.1 kg of BW). One lot of whole blood was isolated with 25% of the total lot dried on d 0, 3, 8, and 12 after collection to create a range in pH. Overall, pigs fed blood meal had improved G:F (P < 0.01) compared to pigs fed the control diet. Similar to Exp. 1, the ammonia concentration of blood meal increased with decreasing pH. Blood meal pH did not influence ADG, ADFI, or G:F (P > 0.21), but pigs fed irradiated blood meal (pH 5.9) had greater ADG and G:F (P < 0.05) than pigs fed nonirradiated blood meal (pH 5.9). In Exp. 3, 300 barrows (17 +/- 6 d of age at weaning) were used to determine the effects of blood meal irradiation source (gamma ray vs. electron beam) and dosage (2.5 to 20.0 kGy) on growth performance of nursery pigs from d 4 to 18 postweaning (8.7 to 13.2 kg of BW). Overall, the mean of all pigs fed blood meal did not differ in ADG, ADFI, or G:F (P > 0.26) compared to pigs fed the control diet without blood meal. Pigs fed irradiated blood meal had a tendency (P < 0.10) for increased G:F compared with pigs fed nonirradiated blood meal. No differences in growth performance were detected between pigs fed blood meal irradiated by either gamma ray or electron beam sources (P > 0.26) or dosage levels (P > 0.11). These studies suggest that pH alone as an indicator of blood meal quality is not effective and irradiation of blood meal improved growth performance in nursery pigs.     

蜂胶HPLC指纹图谱真伪鉴别初探

从蜂胶的功效成分—黄酮类化合物着手,测定了真品蜂胶、杨树胶和搀假蜂胶的HPLC指纹图谱。结果发现:真品蜂胶含有19个特征峰中的18个特征峰,缺6号峰;杨树胶含有19个特征峰中的17个特征峰,缺7号和11号峰;搀了杨树胶的搀假蜂胶,含有6号峰,而7号峰和11号峰的峰面积显著降低。通过上述HPLC指纹图谱可以对蜂胶的真伪进行初步鉴别。     

干酪乳杆菌LC-03的培养条件优化研究

本研究通过单因子优化试验和正交试验对干酪乳杆菌LC-03培养条件和培养基成分进行初步优化。结果表明,干酪乳杆菌LC-03属于兼性厌氧菌,最佳培养基成分为:蛋白胨12g,酵母提取物6g,牛肉膏6g,葡萄糖18g,乙酸钠5g,柠檬酸铵2.15g,Tween801mL,MgSO4.7H2O0.58g,MnSO4.4H2O0.05g,K2HPO42g,水1000mL;最佳培养条件:温度为35℃,培养基起始pH为6.5,接种量为1.0%,培养时间为24h;优化培养后D610nm值达到1.832,活菌数达到3.22×1010CFU/mL。     

Further studies on the development and availability of infective larvae of bovine gastrointestinal trichostrongylids on pasture in eastern Nigeria

The dynamics of pasture populations of infective larvae (L3) of Cooperia, Haemonchus and Trichostrongylus species were studied at Nsukka, eastern Nigeria, during April to November 1984. Six paddocks were contaminated artificially and one was contaminated naturally. Five of the paddocks, P1-P5, were sequentially contaminated with faeces of naturally infected cattle at approximately 4-6-weekly intervals. Paddocks P6 and P7 were repeatedly contaminated every 4-6 weeks artificially and by the naturally infected cattle, respectively. Larval development and survival occurred very readily during the wet season (April-October) but apparently ceased in November at the start of the dry season. Larval migration, however, occurred not only during the rains but also during the first 4 weeks of the dry season. Single contaminations during the rains quickly gave rise to single waves of infestation which also declined rapidly, in spite of the continuously favourable conditions for larval development and survival. The repeated contaminations produced three and four distinct and relatively short-lived larval peaks, respectively, with the first three peaks on both paddocks, namely the May, July and September/October peaks, being coincident. The four waves of herbage infestation on P7, which occurred at approximately 4-5 weekly intervals, were considered to have originated from four separate generations of the three trichostrongylids. However, Trichostrongylus sp. predominated in the first (May) peak while Cooperia and Haemonchus dominated the later peaks.     

Involvement of Plasma Progesterone, Oestradiol-17β and Cortisol in Ovulatory Response to Gonadotropin-releasing Hormone in Dairy Cows with Cystic Follicles

The aim of the present study was to examine the plasma concentrations of progesterone, oestradiol-17beta and cortisol in the cows with cystic follicle and to examine its relationship with the ovulatory response to gonadotropin-releasing hormone (GnRH). Eighty-five post-partum Holstein-Friesian cows with cystic follicles regardless of the presence of corpus luteum were studied. Follicular size, presence of corpus luteum and occurrence of ovulation were checked by palpation per rectum. Blood collection and palpation per rectum were conducted on days 0, 7 and 14. Gonadotropin-releasing hormone was administered at day 7. Plasma concentrations of progesterone, oestradiol-17beta and cortisol were determined. Progesterone concentrations of <3.2, 3.2-4.8 and >4.8 nmol/l were defined as low, intermediate and high, respectively. Sixty-three (74.1%) of 85 cows showed low (<3.2 nmol/l =1 ng/ml) progesterone concentrations on day 0. Only 40 (47.1%) of them showed low-low pattern of progesterone at days 0 and 7. In 27 (31.8%) of them, progesterone concentration had increased by day 14. Of 22 cows having high progesterone concentration (>/=4.8 nmol/l) on day 0, corpus luteum was not detected in 18 cows (21.2%). Only in 10 cows, cystic follicle disappeared after GnRH administration. However, only one of 27 cows in which progesterone pattern was low-low-high at days 0, 7 and 14 experienced ovulation of the cystic follicle. Significantly lower oestradiol-17beta concentration was found on day 7 in cows showing a low-low-low pattern than a low-low-high pattern of progesterone (43.0 +/- 4.6 vs 55.8 +/- 2.8 pmol/l, p < 0.05). There was no significant difference in cortisol concentration on any days (days 0, 7 and 14) between cows showing a low-low-low and low-low-high pattern of progesterone. These results suggest that approximately one-fifth of cows diagnosed to have ovarian cysts possess luteal cysts and that a high oestradiol-17beta concentration at the time of GnRH administration is involved in the subsequent ovulation of the follicle, although ovulated follicle may not be cystic.