牛副结核分枝杆菌LAMP快速检测方法的建立

为建立牛副结核分枝杆菌(MAP)的快速检测方法,本研究采用环介导等温扩增(LAMP)技术,以MAP的IS900基因序列为靶基因设计特异性引物,建立了MAP特异性的LAMP快速检测方法。经反应条件优化,检测结果显示,建立的LAMP检测方法具有良好的特异性,最低见测量为0.53 pg;与其他病原菌无任何交叉反应。临床样品检测与行业标准的符合率为100%。该LAMP检测方法可以用于MAP的常规检疫。     

Prevalence of campylobacter in pigs during fattening; an epidemiological study

Summary

Numerous epidemiological reports implicate foods of animal origin as vehicles of human campylobacteriosis. Pigs are probably an important reservoir of campylobacter and a potential source of human infection. In order to improve our knowledge of the epidemiology of campylobacter in pigs, the prevalence of campylobacter and its contamination of feed were monitored in eight pig farms. Faeces samples of pigs aged 11 and 22 weeks, and samples of rectal, ileal and gastric content at a slaughterhouse were collected for bacteriological examination. On 5 farms, subsequent groups of pigs housed in the same stalls was sampled, too. A selection of the campylobacter isolates was characterized with a genetic typing method (RFLP). More than 85% of the sampled porkers were shown to be intestinal carriers of campylobacter at all stages of fattening. Subsequent groups of pigs housed in the same stalls were all carriers, too. The level of campylobacters in the faeces tended to decrease as the pigs got older. There was no difference in the frequency and level of infection with campylobacter between porkers on different farms. The feeding system (wet feed versus dry pellets) did not seem to influence the prevalence of campylobacter although wet feed gave lower counts of Enterobacteriaceae in the faeces.

RFLP‐typing showed a high diversity of campylobacter strains at each sampling on the farm. Similarities were seen between strains isolated during two subsequent samplings of the same group of pigs, but not between strains isolated on the same farm from subsequent groups of pigs housed in the same stall. This suggests that the piglets were already infected at a young age on the breeding farm.     

赤霉素喷施对红灯甜樱桃果实品质及解剖结构的影响

研究红灯甜樱桃生长期喷施不同浓度(20、60、100 mg·L^-1)赤霉素(GA₃)对果实品质及解剖结构的影响。结果表明,60 mg·L^-1的GA₃处理增加了单果重与纵径。不同浓度GA₃处理对果实横径无显著影响,但降低了果实硬度、可溶性糖、维生素C和可溶性固形物含量,升高了可滴定酸含量,致果实品质总体下降。解剖结构结果表明,GA₃喷施浓度越大,果实角质层与表皮厚度越薄,果肉细胞与表皮细胞越大,硬度越小。建议在生产中减少GA₃的使用,尤其是高浓度(100 mg·L^-1)的GA₃喷施对果实不利。     

甜樱桃矮化砧木矮化机理解剖学研究

本研究以六种甜樱桃砧木为试材，对其根皮率、根导管面积、根导管密度、根导管总面积／木质部面积比值、根木质部面积／横断面积比值、枝皮率、枝导管面积、枝导管平均总面积／木质部平均面积比值和枝木质部面积／横断面积比值等解剖结构进行了比较。试验结果表明不同砧木间各测定值存在较大差异，可以认为根皮率，枝导管密度、根、枝导管总面积／木质部面积比值和根、枝木质部面积／横断面面积比值可以作为预测甜樱桃矮化砧木矮化性的综合指标，为今后甜樱桃矮化砧木的选育提供依据。     

根际促生细菌混合接种对甜樱桃/东北山樱根际微生物区系及根系呼吸的影响

为探讨有益微生物对甜樱桃根系的接种效应,试验采用灌根的方法,研究了从野生东北山樱根际分离筛选出的4株促生细菌,混合接种对甜樱桃/东北山樱根际微生物区系及根系呼吸代谢途径的影响.结果表明:接种促生菌液改变了甜樱桃根际细菌、放线菌及真菌的数量比例,使根际细菌数量显著增加.接种2次,根际细菌增加幅度最大,为对照的3.4倍;接种促生菌液提高了根系活力,接种2次效果最显著,为对照的2.4倍,达到200.4μg/(g·h);接种促生菌液对根系呼吸的基础生化途径与末端氧化电子传递途径运行比例无明显影响,但整体上提高了根系呼吸速率.     

牛副结核病的危害及其防控措施

由副结核分枝杆菌引起牛的副结核病常导致感染牛的慢性增生性肠炎,以及体重、产奶量等生产性能下降,给畜牧业带来了严重的经济损失,目前尚无有效治疗药物。人们常采用不同的检测技术定期对牛群进行监测、淘汰活动带菌期的牛、对牛群进行疫苗接种、采用生物安全防控等措施比较有效地避免了副结核分枝杆菌在牛群中的传播。文章主要从副结核分枝杆菌背景,牛副结核病的危害及防控措施三个方面进行了综述,以期为牛副结核病的防控提供思路。     

Temporal patterns and quantification of excretion of Mycobacterium avium subsp paratuberculosis in sheep with Johne's disease

OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases.     

DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP

OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied.     

Sensitivity and specificity of two serological tests for the detection of ovine paratuberculosis

OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.