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。     

超高效液相色谱-串联质谱法测定猪牛组织中甲基盐霉素残留量

建立超高效液相色谱-串联质谱法测定猪牛组织中甲基盐霉素残留量。猪、牛组织中残留的甲基盐霉素,经乙腈溶液提取,硅胶固相萃取柱净化,超高效液相色谱-串联质谱法测定,外标法定量。甲基盐霉素在5 ng/mL～250 ng/mL的浓度范围内呈现良好的线性关系,甲基盐霉素在猪、牛肌肉、肝脏、肾脏、脂肪中的检测限均为2.5μg/kg,定量限均为5.0μg/kg,猪牛肌肉、肝脏、肾脏、脂肪中5.0 ng/g～100 ng/g添加浓度范围内的回收率均值为71.9%～88.4%,批内批间RSD值均<15%。     

超高效液相色谱-串联质谱法检测牛可食性组织中氟佐隆残留

建立了一种检测牛可食性组织中氟佐隆残留的超高效液相色谱-串联质谱法。牛肌肉、肝脏、肾脏和脂肪样品经90%乙腈溶液提取，高速离心去除蛋白质等杂质，EMR-Lipid柱净化。以0.1%甲酸乙腈溶液和0.1%甲酸水溶液为流动相，在C₁₈色谱柱上分离。在电喷雾正离子(ESI⁺)模式下采用多反应监测模式(MRM)检测，基质匹配标准溶液外标法定量。结果表明：氟佐隆对牛肌肉在1～100 ng/mL浓度范围内，对牛肝脏、肾脏和脂肪在1～500 ng/mL浓度范围内均呈现良好的线性关系，决定系数(R²)均大于0.990;在4种牛可食性组织中氟佐隆的检测限均为5μg/kg,定量限均为10μg/kg。氟佐隆在牛肌肉10～400μg/kg、牛肝脏和牛肾脏10～1000μg/kg、牛脂肪10～14000μg/kg空白添加浓度范围内的回收率为72.0%～98.6%;批内与批间相对标准偏差均小于15%。该方法具有简便快捷、选择性好、定性准确，重复性好等特点，可以满足上述组织中氟佐隆残留检测的要求。     

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莫昔克丁的残留

建立了一种可准确定性定量检测4种牛可食性组织中莫昔克丁残留的液相色谱-三重四极杆/线性离子阱(LC-Qtrap)复合质谱分析技术。牛肌肉、肝脏、肾脏和脂肪样品经乙腈提取,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,在BEH C18色谱柱上实现分离,在电喷雾正离子(ESI+)模式下,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,基质匹配标准溶液外标法定量。结果表明:莫昔克丁在牛肌肉和牛肾脏1~100 ng/mL的基质匹配浓度范围内;在牛肝脏1~200 ng/mL的基质匹配浓度范围内和在牛脂肪1~1000 ng/mL的基质匹配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种牛可食性组织中莫昔克丁的定量限均为2μg/kg。莫昔克丁在牛肌肉2~40μg/kg、牛肝脏2~200μg/kg、牛肾脏2~100μg/kg和牛脂肪2~1000μg/kg添加浓度水平上的回收率范围为65.6%~115%;批内与批间相对标准偏差均小于15%。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莫昔克丁残留检测的要求。     

高效液相色谱法测定牛羊组织中碘醚柳胺残留量

建立高效液相色谱荧光检测法测定牛羊组织中碘醚柳胺残留量。牛羊组织中残留的碘醚柳胺,经乙腈-丙酮溶液提取,阴离子交换固相萃取柱净化,高效液相色谱-荧光法测定,外标法定量。碘醚柳胺在10～5000 ng/mL的浓度范围内呈现良好的线性关系,碘醚柳胺在牛羊肌肉、肝脏、肾脏、脂肪中的检测限均为2.5 μg/kg,定量限均为5 μg/kg,牛羊肌肉、肝脏、肾脏、脂肪中15～500 ng/g添加浓度范围内的回收率为77.4%～93.8%,批内批间RSD值均<15％。     

高效液相色谱-串联质谱法检测牛组织和奶中咪多卡残留的研究

建立了一种检测牛组织和牛奶中咪多卡残留检测的高效液相色谱-串联质谱法。牛组织(肌肉、肝脏、肾脏、脂肪)和奶在NaAc缓冲体系中酶解,经HCl溶液提取,WCX固相萃取柱净化,以0.3 %甲酸水溶液(含20mM甲酸铵)和0.3 %甲酸乙腈为流动相进行梯度洗脱,在HILIC色谱柱上分离,在电喷雾正离子(ESI+⁾模式下,用多反应监测(MRM)模式检测,同位素内标法定量。结果表明：咪多卡在2.5～1000 ng/mL的浓度范围内呈现良好线性关系,相关系数(R2⁾大于0.99;咪多卡在牛组织和奶中的检测限均为10 μg/kg,定量限均为20 μg/kg;咪多卡在牛组织和奶中20～4000 μg/kg添加浓度水平上的回收率在70.9～109%范围内;批内RSD在0.55~9.59%之间,批间RSD在2.21~12.1%之间。该方法具有灵敏度高、定量准确,重复性好等特点,可以满足牛组织和奶中咪多卡残留检测的要求。     

高效液相色谱-串联质谱法检测猪可食性组织中加米霉素残留

为了建立一种可靠、灵敏的检测猪可食性组织中加米霉素残留量的高效液相色谱-串联质谱方法,本试验将猪组织(肌肉、肝脏、脂肪和肾脏)经绞碎、均质后,用乙腈提取,乙腈饱和正己烷去脂,固相萃取柱净化,Kinetex C18色谱柱梯度洗脱,在电喷雾正离子下以多反应监测模式检测,内标法定量。结果显示,该方法的检测限和定量限分别为3μg/kg和5μg/kg;加米霉素在5～1 000μg/kg浓度范围内,峰面积与浓度的线性关系良好;猪肌肉、肝脏和脂肪在5、50、100μg/kg和200μg/kg四个添加浓度的平均回收率为70.20%～90.75%,批间变异系数≤11.94%;猪肾脏在5、150、300μg/kg和600μg/kg四个添加浓度的平均回收率为66.69%～87.46%,批间变异系数≤9.95%。结果表明该方法灵敏、准确,可满足兽药残留检测的要求。     

牛组织中氨丙啉残留的HPLC检测方法研究

建立了牛组织中氨丙啉残留检测的高效液相色谱法.试样中的氨丙啉经乙腈提取后旋转蒸干,用磷酸盐缓冲液溶解,过HLB固相萃取小柱净化,高效液相色谱法进行测定.结果表明,在0.20～20.0 μg/mL范围内,氨丙啉浓度与峰面积呈良好线性关系(r2=0.9999);在牛的肌肉、肝脏、肾脏和脂肪中,氨丙啉的检测限为100 μg/kg,在牛的肌肉、肝脏、肾脏中定量限为250 μg/kg,在牛的脂肪中定量限为200 μg/kg.在不同添加浓度下测定,氨丙啉的平均回收率为60％ ～ 110％,批内和批间变异系数均在15％以内.本方法可快速测定牛组织中氨丙啉残留量,且操作简单、灵敏度高、结果准确性高,能满足日常检测需要.     

高效液相色谱-串联质谱法检测牛组织中双氯芬酸钠残留的研究

为了测定牛肌肉、肝脏、肾脏组织中双氯芬酸钠的残留量,研究建立了一种用于该检测的高效液相色谱-串联质谱(LC-MS/MS)方法。试验将样品中加入乙腈提取液重复提取2次,超声波辅助提取,提取液中加入C₁₈、PSA混合吸附剂进行净化。色谱柱采用C₁₈反相柱对药物进行分离,洗脱液采用乙腈-0.1%甲酸水进行等度洗脱,配备电喷雾离子源,药物在正离子模式下电离,多反应监测模式(MRM)分段扫描测定检测结果,并采用内标法定量分析。结果表明,双氯芬酸在0.5～100μg/L浓度范围内有良好的线性关系,相关系数大于0.99;在牛的肌肉、肝脏和肾脏组织中检测限(LOD,S/N=3)为0.5μg/kg,定量限(LOQ,S/N=10)为1.0μg/kg。在3个浓度添加水平下,平均回收率为97.7%～114%,日内变异系数0.5%～4.7%,日间变异系数为0.9%～7.5%。该方法操作简便、结果准确,满足兽药残留检测方法规定要求,运用于快速准确测定牛肌肉、肝脏、肾脏等组织中双氯芬酸钠的残留。     

高效液相色谱-串联质谱法检测鸡蛋和鸡组织中恩诺沙星、环丙沙星残留的研究

为建立一种高效液相色谱-串联质谱法检测鸡蛋和鸡组织(肝脏、肾脏、肌肉、脂肪)中恩诺沙星、环丙沙星的残留,鸡蛋和鸡组织(肝脏、肾脏、肌肉、脂肪)经酸化乙腈提取,正己烷除脂净化,以0.1%甲酸水溶液和乙腈为流动相进行梯度洗脱,同位素内标法定量。结果显示:恩诺沙星、环丙沙星均在线性范围内呈现良好线性关系,相关系数(R2)均大于0.99;恩诺沙星、环丙沙星在组织中的检测限为1μg/kg,定量限为5μg/kg;两种药物的平均回收率范围为75.5%～106%,批内、批间变异系数均小于11.2%。研究表明,该方法样品前处理简单,灵敏度高,适用同时检测鸡组织(肝脏、肾脏、肌肉、脂肪)和鸡蛋中恩诺沙星、环丙沙星残留。     

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.     

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.     

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.     

Micronutrient Content of Certain Tropical Conventional and Unconventional Feed Resources of Southern India

The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies.     

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.     

Bovine Mastitis in Selected Areas of Southern Ethiopia

A study on bovine mastitis, designed to determine the causal agents, prevalence of infection and impact of risk factors in three cattle breeds, was conducted in selected areas of southern Ethiopia. A total of 307 lactating and non-lactating cows, of which 162 were indigenous Zebu, 85 Jersey and 60 Holstein-Friesian, were examined by clinical examination and the California mastitis (CMT) test. Of these, 40.4% were positive by CMT and bacteriology for clinical or subclinical mastitis, with prevalence rates of 37.1% and 62.9%, respectively. Out of 1133 quarters examined, 212 (18.7%) were found to be infected, 83 (39.2%) clinically and 129 (60.8%) subclinically. The prevalence of mastitis was significantly higher in Holstein-Friesian than in indigenous Zebu, in non-lactating cows than in lactating cows, in the early lactation stage than in the mid-lactation stage, in cows with lesions and/or tick infestation on skin of udder and/or teats than in cows without this factor, and in the wet season than in the dry season. Mastitis increased with parity number (R = 0.9). Of 248 CMT and clinically positive udder quarter samples analysed microbiologically, 212 were culturally positive for known mastitis pathogens and 36 were negative. Of the 199 positive samples, Staphylococcus accounted for 39.2%, Streptococcus for 23.6%, coliforms for 14.1%, Micrococcus and Bacillus species for 8.0% each and Actinomyces or Arcanobacterium (Corynebacterium) for 7.0%. It was concluded that there was a high prevalence of clinical and subclinical mastitis, mainly caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli, in this study area.     

不同日粮氮水平对山羊氮代谢和微生物蛋白质合成的影响

本试验旨在探讨不同日粮氮水平对内蒙古白绒山羊氮代谢和微生物蛋白质合成(MCP)的影响,以达到提高氮利用率,减少环境污染和饲料资源的浪费。试验选用9只体况良好、体重为(43.83±2.95)kg,装有瘤胃瘘管的内蒙古白绒山羊,按体重随机分为3组,每组3只。日粮分为低氮7.5%、中氮10.5%、高氮13.5%3个氮水平。试验使用常规试验方法分析测定粪尿血液等生化指标,通过全收尿法和尿嘌呤衍生物法估测MCP。结果表明:随着氮水平增加,氨氮(NH3-N)浓度、氮摄入量、尿氮排出、尿中尿素氮(UUN)和MCP显著增加(P0.05),干物质采食量(DMI)显著降低(P0.01);沉积氮占总摄入氮的比例在中氮组最高,MCP在高氮组最高。综上所述,适当降低日粮氮水平可减少粪尿氮排放,提高反刍动物氮利用率。     

Microsporidial keratopathy in two dogs

A microsporidial keratopathy is described in two dogs. Both dogs presented with a unilateral stromal keratopathy characterized by multifocal coalescing opacities, and the diagnosis was made on histopathologic examination of keratectomy specimens. Transmission electron microscopy (TEM) on formalin‐fixed, paraffin‐embedded corneal tissue was performed in one dog, and the morphologic features were consistent with Nosema species infection. Both dogs were initially diagnosed and treated by superficial keratectomy. One dog received additional antifungal medication and underwent a penetrating keratoplasty following local recurrence two years later. No other systemic lesions attributable to the microsporidial infection were identified clinically. The clinical and diagnostic pathology findings, treatment, and follow‐up are discussed.