液相色谱-串联质谱法检测牛奶中4种阿维菌素类药物残留量的研究

建立了一种可同时检测牛奶中4种阿维菌素类药物(阿维菌素、伊维菌素、多拉菌素和埃谱利诺菌素)残留的液相色谱-串联质谱方法(LC-MS/MS)。牛奶样品经乙腈提取,高速离心去除蛋白质等杂质,C18柱净化。以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相进行洗脱,流速0.4 m L/min,采用基质匹配标准溶液外标法定量。结果表明:阿维菌素、伊维菌素、多拉菌素和埃谱利诺菌素在0.5~100 ng/m L浓度范围内均呈现良好的线性关系,相关系数(R2)均大于0.996;4种阿维菌素类药物的定量限均为0.5μg/kg。阿维菌素在0.5~5μg/kg添加浓度范围内,伊维菌素在0.5~20μg/kg添加浓度范围内,多拉菌素在0.5~30μg/kg添加浓度范围内,埃谱利诺菌素在0.5~40μg/kg添加浓度范围内,回收率为66.4%~120%;批内与批间相对标准偏差均小于20%。该方法具有简便快速、灵敏度高、定性准确,重复性好等特点,可以满足牛奶中4种阿维菌素类药物残留检测的要求。     

HPLC-QTOF和HPLC-MS/MS筛查与确证兽药中非法添加氧氟沙星等15种喹诺酮类药物

采用高效液相色谱-四级杆-飞行时间质谱(HPLC-QTOF)筛查并确证1批监督抽检的兽药硫酸黏菌素注射液中的未知添加物,并建立液相色谱-三重四级杆质谱法(HPLC-MS/MS)定量检测20种兽药中15种该类化合物的方法。采用HPLC-QTOF筛查兽药非法添加物时,发现1批硫酸黏菌素注射液中含有未知添加物,与建立的一级谱库中氧氟沙星匹配分数很高,疑似为氧氟沙星。通过二级质谱碎片离子的推导及与对照品二级质谱比对,确证其为氧氟沙星。采用HPLC-MS/MS建立了定量检测20种常见兽药中非法添加氧氟沙星、恩诺沙星、沙拉沙星、达氟沙星、依洛沙星、洛美沙星、麻保沙星、诺氟沙星、环丙沙星、培氟沙星、司帕沙星、二氟沙星、奥比沙星、恶喹酸、氟甲喹共15种喹诺酮类化合物的方法,15种喹诺酮类化合物在各自的线性范围内均呈现良好的线性关系,添加回收率为85.0%～100.0%,相对标准偏差(RSD,n=3)均不高于15.0%。通过检查该样品中的非法添加物,建立了简便、快速、灵敏的定性定量检测方法,为打击当前兽药中非法添加化合物建立技术标准提供参考,为兽药处方外非法添加筛查与确证提供思路。     

液相色谱-三重四级杆/线性离子阱复合质谱技术检测牛可食性组织中吡利霉素的残留

建立了一种可准确定性定量检测4种牛可食性组织中吡利霉素残留的液相色谱-三重四级杆/线性离子阱（LC-Q-trap MS）复合质谱分析技术。牛肌肉、脂肪、肾脏和肝脏样品经5%甲酸乙腈提取,captiva EMR-Lipid柱净化。以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相,经Kinetex F5 色谱柱分离后,用多反应监测联合信息依赖性采集与增强子离子扫描（MRM-IDA-EPI）模式检测,基质匹配标准溶液校正,外标法定量。结果表明：吡利霉素于牛肌肉和脂肪中在1～100 ng/mL的浓度范围内,于牛肾脏和肝脏中在1～400 ng/mL浓度范围内均呈现良好的线性关系,关系数（R2）均大于0.990;在4种空白组织中吡利霉素的检测限均为5 μg/kg,定量限均为1相0 μg/kg。吡利霉素在牛肌肉和牛脂肪10～200 μg/kg、牛肾脏10～800 μg/kg,以及牛肝脏10～2000 μg/kg添加浓度范围内的回收率范围在60.2%～101%之间;批内与批间相对标准偏差均小于13%。该方法具有简便快速、灵敏度高、定性准确,重复性好等特点,可以满足上述组织中吡利霉素残留检测的要求。     

液相色谱-三重四极杆/线性离子阱复合质谱技术检测牛可食性组织中吡利霉素的残留

建立了一种可准确定性定量检测4种牛可食性组织中吡利霉素残留的液相色谱-三重四极杆/线性离子阱(LC-Q-trap MS)复合质谱分析技术。牛肌肉、脂肪、肾脏和肝脏样品经5%甲酸乙腈提取,captiva EMR-Lipid柱净化。以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相,经Kinetex F5色谱柱分离后,用多反应监测联合信息依赖性采集与增强子离子扫描(MRM-IDA-EPI)模式检测,基质匹配标准溶液校正,外标法定量。结果表明:吡利霉素于牛肌肉和脂肪中在1～100 ng/mL的浓度范围内,于牛肾脏和肝脏中在1～400 ng/mL浓度范围内均呈现良好的线性关系,相关系数(R~2)均大于0.990;在4种空白组织中吡利霉素的检测限均为5μg/kg,定量限均为10μg/kg。吡利霉素在牛肌肉和牛脂肪10～200μg/kg、牛肾脏10～800μg/kg,以及牛肝脏10～2000μg/kg添加浓度范围内的回收率范围在60.2%～101%之间;批内与批间相对标准偏差均小于13%。该方法具有简便快速、灵敏度高、定性准确,重复性好等特点,可以满足上述组织中吡利霉素残留检测的要求。     

QuEChERS-液相色谱串联质谱法测定动物源性食品中氟喹诺酮类残留量

建立了用QuEChERS-液相色谱串联质谱法测定禽肉、禽蛋和牛奶中15种氟喹诺酮类药物残留的检测方法。样品经酸化乙腈提取后,经QuEChERS试剂净化,液相色谱串联质谱测定,内标法定量。结果表明,牛奶和禽肉的检出限为2.0 ng/g,定量限为4.0 ng/g,禽蛋的检出限为3.0 ng/g,定量限为4.0 ng/g。在2.0~100 ng/mL的添加浓度范围内线性良好,线性相关系数(r)大于0.995,牛奶、禽肉以及禽蛋三种基质在添加浓度为4.0~50.0 ng/g范围时,回收率为61%~105.2%,批内、批间精密度均小于15%。该方法简便、快速、准确,适用于大批量动物源性样品中氟喹诺酮的快速检测。     

猪肉组织中四环素类药物残留检测——高效液相色谱-串联质谱法研究

建立了猪肉组织中四环素类药物残留检测的高效液相色谱-串联质谱方法.色谱条件为色谱柱为XBridge C18柱(150 mm×2.1 mm,5 μm),流动相为0.3%甲酸乙腈溶液 0.3%甲酸水溶液;柱温 20 ℃,流速0.2 mL/min;进样量20 μL.质谱条件为电喷雾离子源(ESI );多反应监测(MRM)方式采集.外标法定量.结果表明,四环素、土霉素和金霉素在10～400 ng/mL浓度范围内呈现良好线性,相关指数R2均大于0.999;方法检测限为5 ng/g,定量限为10 ng/g.50、100和200 ng/g三个添加浓度的平均回收率为72.3%～94.6%,批内批间RSD均小于20%.     

高效液相色谱-四级杆/线性离子阱质谱法筛查兽药中非法添加物的研究

建立了一整套利用高效液相色谱-四级杆／线性离子阱质谱对兽药中非法添加物进行筛查的方法。首先,通过比较样品与阴性对照的差异,采用Q3全扫描方式获取靶目标化合物的母离子质荷比（214．2Da）,再对该化合物的质谱参数、色谱分离条件等进行逐一优化,最后利用线性离子阱质谱的“多反应监测触发增强子离子扫描”功能对靶目标化合物与相应标准物质进行谱库匹配分析。综合靶目标化合物的保留时间（6．92min）、离子质荷比、特征碎片离子及谱库匹配度（93．2）等参数,确证了该品牌的部分兽药中非法添加了氯丙那林。本研究为兽药中非法添加物筛查及定性确证提供了一个有效的思路与方法。     

液相色谱-串联质谱法检测牛奶中β-内酰胺类药物残留的研究

β-内酰胺类药物主要有青霉素G、青霉素V、阿莫西林、羧苄西林、氨苄西林、苯唑西林、氯唑西林、萘夫西林、头孢喹肟、头孢氨苄、头孢拉定、头孢唑啉和头孢哌酮13种药物。牛奶中β-内酰胺类药物经乙腈提取,正己烷脱脂后,经C18净化小柱净化,取适量上清液过0.22μm滤膜后,供液相色谱-串联质谱仪测定。经试验,浓度为5、10、50、100、200和500 ng/m L的基质匹配系列混合标准溶液,离心过滤后上机测定。以特征离子质量色谱峰面积为纵坐标,标准溶液浓度为横坐标,绘制标准曲线,13种β-内酰胺类药物在5-500 ng/m L的浓度范围内呈现良好的线性关系,R2均大于0.990。牛奶在2-50μg/kg添加浓度范围内,本方法的回收率为70%-120%;批内、批间RSD均小于20%。检测限为1μg/kg,定量限为2μg/kg。     

液相色谱-三重四极杆/线性离子阱复合质谱技术检测牛奶中莫昔克丁残留

建立了牛奶中莫昔克丁残留检测的液相色谱-三重四极杆/线性离子阱复合质谱技术(LC-QTRAP)。样品用乙腈提取后,用C18固相萃取柱净化,上机测试。液相色谱条件:色谱柱为BEH C18(50 mm×2.1 mm,1.7μm),流动相为0.1%甲酸水溶液+0.1%甲酸乙腈溶液,梯度洗脱,流速为0.4 mL/min,柱温为30℃,进样量为5μL。质谱条件:电喷雾正离子源(ESI~+),采用多反应监测-信息依赖扫描-增强子离子扫描(MRM-IDA-EPI)方式采集。结果表明:在牛奶中,莫昔克丁在1～80 ng/mL基质匹配标准溶液浓度范围内均呈现良好的线性关系,相关系数(R~2)大于0.990,定量限为2μg/kg。牛奶中莫昔克丁在2、40和80μg/kg三个添加浓度平均回收率都在60%～120%之间,批内、批间RSD均小于15%。该方法简便快捷、定性准确、灵敏度高,回收率和精密度满足残留检测要求。     

鸡蛋及牛奶中四环素类药物残留量的液相色谱-串联质谱测定法

建立了一种测定鸡蛋及牛奶中四环素类药物残留 液相色谱-串联质谱的方法。样品经Mcllvaine-Na2EDTA缓冲液提取,经快速滤纸过滤,并经加脱脂棉的HLB固相萃取柱净化,洗脱液氮气吹至小于1 mL,加30%甲醇至1mL并混匀,液相色谱-串联质谱法检测,基质匹配标液外标法定量。结果表明,四环素类药物在5 μg/kg~100 μg/kg的浓度范围内呈现良好的线性关系。在鸡蛋及牛奶中四环素类药物的检测限均为2 μg/kg,定量限均为5 μg/kg。在LOQ~2MRL(多西环素为LOQ-10LOQ)添加浓度范围内,四环素类药物的回收率均在81.2~110.0%之间,批内批间变异系数均小于11.4%。该方法各项技术指标均能满足残留检测要求,且方法的重现性良好,满足国内外兽药残留检测的相关规定。     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

Feline intracranial neoplasia: retrospective review of 160 cases (1985-2001)

The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

A brief history of the prevention of infectious diseases by immunisations

Infectious diseases have always been a terrible scourge for humans. The appearance of these plagues, as they were called without distinction, was generally connected to various conditions: asters, climatic changes or religious reasons. The concept of contagious, and then infectious, diseases came slowly. Variolation, i.e. transmission of ‘virulent’ matter to induce a natural disease and the immunity against it, was brought from Constantinople to England by Lady Montague, in 1721. This ‘variolation’ technique was also often performed in veterinary medicine against diseases like sheep-pox or pleuropneumonia. As ‘vaccination’ is the term generally accepted for ‘immunisation’, variolation can be the word designating such a technique. The second period of the history of immunisation began, in 1880, with the studies of Pasteur and his collaborators. A great number of bacterial vaccines were developed: dead, live but attenuated or only parts of pathogens. The viruses were produced in animals, then in eggs and at last, in tissue cultures. Second generation vaccines appeared with genetic engineering: recombinant vaccines, vector vaccines, nucleic acids vaccines, and markers vaccines, among others. These novel technologies can permit the development of new ones and improve the quality of the vaccines already existing.