抗大肠杆菌的IgY的分离制备及功能性测定

以大肠杆菌（Ｅ．ｃｏｌｉ）为抗原，对蛋鸡进行免疫，其ＩｇＹ的活性提高３２倍，并能有效地抑制Ｅ．ｃｏｌｉ的生长。功能性实验表明，６０℃以下，ｐＨ４以上免疫球蛋白活性基本没有损失，在低浓度胃蛋白酶存在下，仍具有较高的活性。     

从市售蔬菜和熟肉制品中检测出O157:H7大肠杆菌

肠出血性大肠杆菌（ＥｎｔｒｅｏｈｅｍｏｒｒｈａｇｉｃＥｓｃｈｅｒｉｃｈｉａＣｏｌｉ,ＥＨＥＣ）是Ｒｉｌｅｙ等于１９８２年首次报告的,〔１〕血清型为Ｏ１５７：Ｈ７。大肠杆菌Ｏ１５７：Ｈ７引起的食源性疾患死亡率较高。〔２,４〕近年来,大肠杆菌Ｏ１５７：Ｈ...     

产细菌素植物乳杆菌菌株的筛选及其细菌素生物学特征研究

从甘蓝泡菜中分离到６９株乳酸杆菌,通过琼脂点扩散交叉拮抗试验,筛选出３株有明显抑菌活性代谢产物的植物乳杆菌。排除酸、过氧化氢等干扰因素后,离心发酵液对指示菌Ｌａｃｔｏｂａｃｉｌｕｓｐｌａｎｔａｒｕｍ９６Ｄ仍有抑菌作用,用胰蛋白酶对其透析液处理后活性丧失,说明它们产生的抑菌物质是细菌素。以Ｇ８菌株为试材,对其细菌素类物质的产生及细菌素粗提物进一步研究,发现在对数末期其抑菌活性最高,对热相对稳定（１００℃,２０ｍｉｎ）,易被胰蛋白酶、蛋白酶Ｋ和胃蛋白酶失活,显示活性的ｐＨ值范围为４．０～５．５,粗提液表现为不仅抗明串株菌属、片球菌属、乳杆菌属的一些菌株,而且抗一些非乳酸菌的革兰氏阳性菌,但对大肠杆菌（Ｅ．ｃｏｌｉ）等革兰氏阴性菌没有任何抑制作用。说明Ｇ８菌株产生的是一类抗广谱革兰氏阳性菌的细菌素     

几丁质酶基因在烟草栽培品种中的表达

利用多聚酶链式反应（ＰＣＲ）对来源于一种生物杀虫剂（ｂａｃｕｌｏｖｉｒｕｓ）的几丁质酶基因进行体外扩增，并插入载体质粒ｐＲＯＫ２中，然后转化大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａｃｏｌｉ）ＸＬ１Ｂｌｕｅ。用重组质粒ｐＲＯＫ２ＤＮＡ转化植物转基因载体——土壤农杆菌（Ａｇｒｏｂａｃｔｅｒｉａｔｕｍｅｆａｃｉｅｎｓ）ＬＢＡ４４０４菌株。借助于土壤农杆菌侵染烟草叶圆片，将目的基因导入。通过组织培养诱导生芽和生根，获得烟草再生植株。利用含卡那霉素的培养基对再生烟株进行初步筛选。进一步ＰＣＲ和Ｗｅｓｔｅｒｎ印迹检测结果表明：转基因烟草中有几丁质酶基因和蛋白的表达。初步检测结果表明：转基因烟草具有较高的几丁质酶活性。     

肠出血性大肠杆菌(EHEC)流行趋势(综述)

肠出血性大肠杆菌（Ｅｎｔｅｒｏ－ｈｅｍｏｒｒｈａｇｉｃＥ．ｃｏｌｉ,ＥＨＥＣ）是指能引起人的出血性肠炎的一群大肠埃希氏菌,以Ｏ１５７：Ｈ７血清型菌株为主,还包括Ｏ１５７：ＮＭ,Ｏ２６：Ｈ１１,Ｏ１１１：Ｈ８,Ｏ１２５：ＮＭ,Ｏ１２１：Ｈ１９,Ｏ４：Ｎ...     

酶法生产L—天冬氨酸之研究

本文对利用Ｅ．ｃｏｌｉＡＳ１．８８１细胞内之Ｌ－Ａａｐ酶，以反丁烯二酸及氨为底物转化生产Ｌ－天冬氨酸进行了系统的研究，试验结果表明该酶反应转化率高，Ｌ－Ａｓｐ收率亦较高，Ｌ－Ａｓｐ对反丁烯二酸总收率可达９０％以上。     

牛乳均质效果的测定

牛乳均质效果的测定ＤｅｔｅｃｔｉｏｎｏｆＭｉｌｋＨｏｍｏｇｅｎｉｚａｔｉｏｎＥｆｅｃｔ赵平ＺｈａｏＰｉｎｇ（利乐包装（昆山）有限公司）（ＴｅｔｒａＰａｋ（Ｋｕｎｓｈａｎ）ＣＯ．，Ｌｔｄ．）１概述均质是将乳中脂肪球打碎并使其均匀分布于乳中，有效的均质能...     

肠出血性大肠艾希氏菌O157：H7（综述）

肠出血性大肠艾希氏菌Ｏ１５７：Ｈ７（综述）冉陆卫生部食品卫生监督检验所（１０００２１）１９７５年从一位患出血性腹泻的加利福尼亚妇女的粪便中首次分离到大肠艾希氏菌Ｏ１５７：Ｈ７（ＥｓｃｈｅｒｉｃｈｉａｃｏｌｉＯ１５７：Ｈ７）。１９８２年，Ｒｉｌｅｙ［１...     

防腐抑菌剂的进展

１０－１５７及其对策１９９６年日本大规模暴发肠内出血性大肠杆菌疾病,从日本大阪府数十所小学和保育学校儿童开始,病例报告遍及全日本,患者累计９４５１人,死亡１２人,引起全日本恐慌和世界各国的严重关注。为此,日本厚生省提高了对食品工厂执行危害和关键控制点（ＨＡＣＣＰ）的要求,并将该病正式列为法定报告的传染病。肠内出血性大肠杆菌（Ｅｎｔｅｒｏ－ｈｅｍｏｒｒｈａｇｉｃＥ．Ｃｏｌｉ,简称ＥＨＥＣ）有许多菌株组成,包括Ｏ１５７：Ｈ７,Ｏ１５７：ＮＭ,Ｏ１２５：ＮＭ,Ｏ１４５：ＮＭ,等菌株,而以Ｏ１５７：Ｈ７…     

二种重组α-乙酰乳酸脱羧酶在啤酒生产中降低双乙酰的作用

利用已构建的含有短芽胞杆菌（Ｂａｃｉｌｌｕｓｂｒｅｖｉｓ）和产气肠杆菌（Ｅｎｔｅｒｏｂａｃｔｅｒａｅｒｏｇｅｎｅｓ）、α -乙酰乳酸脱羧酶（α -ａｃｅｔｏｌａｃｔａｔｅｄｅｃａｒｂｏｘｙｌａｓｅ,α -ＡＬＤＣ）基因的工程菌株 ,并使它们分别在大肠杆菌中高效表达 ,获得重组α -ＡＬＤＣ。在实验室 ,用２Ｌ体积的麦芽汁进行啤酒生产试验 ,添加２种重组α -ＡＬＤＣ后 ,使啤酒中的双乙酰含量快速下降到或始终保持在０．１ｍｇ／Ｌ以下。证明得到的重组α -ＡＬＤＣ能有效地降低啤酒中双乙酰含量。这２种重组α -ＡＬＤＣ可进一…     

Efficacy of an Escherichia coli J5 mastitis vaccine in an experimental challenge trial.

An Escherichia coli (O111:B4) J5 bacterin was tested for efficacy in reducing IMI and severity of clinical coliform mastitis in an experimental challenge trial. Ten cows were immunized at drying off, 30 d after drying off, and at calving. Ten control cows were not immunized. Right front quarters of all cows were infused with a heterologous strain of E. coli approximately 30 d after calving. Vaccinated cows had lower bacterial counts in milk and lower rectal temperatures than unvaccinated controls following intramammary challenge. Milk production and DMI were more depressed in controls than in vaccinated cows. Milk SCC did not differ between experimental groups. Mean serum IgG titer to whole cell E. coli J5 was significantly greater in vaccinated than in unvaccinated cows at 30 d after drying off, day of challenge, and 7 d postchallenge. Milk IgG titer to E. coli J5 was higher at challenge in vaccinated than in control cows. Vaccination with the E. coli J5 bacterin did not prevent IMI but did reduce severity of clinical signs following intramammary experimental challenge with a heterologous E. coli strain.     

Efficacy of immunization with ferric citrate receptor FecA from Escherichia coli on induced coliform mastitis

The effects of immunization with the ferric citrate receptor FecA on antibody responses and on experimentally induced mastitis following intramammary challenge were investigated. Twenty-one cows were assigned to seven blocks of three cows based on expected parturition. Cows within block were randomly assigned to one of three treatments: 1) FecA immunization, 2) Escherichia coli J5 immunization, and 3) unimmunized controls. Challenge was by infusion of approximately 60 cfu of E. coli 727 into one uninfected mammary gland between 13 and 31 d after parturition. Cows within block were challenged on the same day. Cows immunized with FecA had higher immunoglobulin (Ig)G titers against FecA in serum and in mammary secretions at calving, immediately before challenge, and 7 d after challenge than did cows immunized with E. coli J5 or control cows. Immunization with FecA also increased IgG titers against whole-cell E. coli 727 in serum and in mammary secretions at calving. Serum IgM titers against FecA were higher in FecA immunized cows than in other treatment groups immediately before challenge. Bacterial counts in milk, duration of bacterial isolation in milk, rectal temperature, and milk somatic cell counts following intramammary challenge were similar among treatments. Milk production and dry matter intake did not differ among treatments. The ferric citrate receptor FecA was immunogenic in cows, but immunization had minimal effect on the clinical severity of experimentally induced E. coli mastitis.     

Effect of immunoglobulin G from cows immunized with ferric citrate receptor (FecA) on iron uptake by Escherichia coli

The effects of immunoglobulin (Ig) G from cows immunized with the ferric citrate receptor (FecA) on iron uptake by Escherichia coli were investigated. Receptor FecA was purified from E. coli UT5600/pSV66. Cows were immunized with 400 microg purified FecA three times at 21 d intervals during late lactation and the nonlactating period. Immunoglobulin G was purified by protein G affinity chromatography from colostral whey from cows immunized with FecA and from unimmunized control cows. The purified IgG from FecA immunized cows had higher IgG titers against FecA compared with control IgG. Fifteen E. coli isolated from intramammary infections and E. coli UT5600/pSV66 were grown in an iron-depleted medium containing 1 mM citrate to induce FecA. The bacterial cells were mixed with 0, 2, and 4 mg/ml purified IgG, and 55Fe was added to the assay. After 5, 10, and 15 min incubations at 37 degrees C, samples were passed through 0.45-pm pore size filters. Filters were washed with saline three times, and the radioactivity of 55Fe taken up by the bacterial cells on the filters was measured by a liquid scintillation counter. The measurements were expressed as numbers of 55Fe atoms per colony-forming unit and transformed to log10. The assay was repeated three times for each isolate in a partially balanced incomplete block design. The presence of IgG decreased 55Fe uptake by E. coli mastitis isolates and E. coli UT5600/pSV66. Anti-FecA IgG reduced 55Fe uptake by E. coli greater than IgG from unimmunized cows.     

Opsonic activity of bovine serum and mammary secretion after Escherichia coli J5 vaccination.

Six pairs of cows were used to determine the effects of immunization with an Escherichia coli (O111:B4) J5 bacterin on in vitro opsonization of a smooth heterologous strain of E. coli. One cow in each pair was either immunized with the vaccine or sham-immunized at drying off, 30 d after drying off, and at calving. Opsonizing bacteria with serum collected from vaccinated cows 21 d after calving resulted in higher mean number of intracellular bacteria per phagocytosing neutrophil than opsonizing bacteria with serum collected from control cows. Phagocytic parameters using serum collected at drying off and calving did not differ between treatment groups. A trend for enhanced opsonic activity of colostrum from vaccinates was noted. Enhanced opsonization by serum from vaccinated cows coincided with higher serum IgM titer to E. coli J5 whole cell antigen compared with controls. Serum IgG titers to E. coli J5 did not differ between groups. Colostrum IgG titers to E. coli J5 were greater at calving in vaccinated than in control cows. Colostrum and milk collected 21 d after calving from vaccinated cows had higher IgM titers to E. coli J5 than did mammary secretions from control cows. Numbers of intracellular bacteria per phagocytizing neutrophil were correlated positively with IgM titers to E. coli J5 in both serum and colostrum.     

A comparison of two commercially available Escherichia coli J5 vaccines against E. coli intramammary challenge

The efficacy of two commercially available Escherichia coli J5 bacterins was investigated. Jersey cows were randomly assigned to one of three treatment groups: 1) unvaccinated controls, 2) vaccinated with J.VAC (Merial Limited, Athens, GA), and 3) vaccinated with J5 bacterin. All cows were vaccinated at drying off and at 2 wk before anticipated calving. Cows that were vaccinated with the J5 bacterin also received a third immunization at calving. One quarter of each cow was challenged with approximately 64 cfu of E. coli at 14 to 30 d postcalving. Immunization by either vaccine did not influence the severity of coliform mastitis; however, the mean number of colony-forming units of E. coli recovered from challenged quarters was significantly lower for immunized cows than for control cows at 144 h postchallenge. Serum and mammary secretion immunoglobulin (Ig)G, IgG1, and IgG2 titers against E. coli J5 whole-cell antigens were enhanced in vaccinated cows. Serum and mammary secretion IgM were not different among treatment groups. Somatic cell counts in milk from challenged quarters, rectal temperatures, and the clinical status of cows following intramammary challenge were not different among treatment groups.     

Intramammary challenge with Escherichia coli following immunization with a curli-producing Escherichia coli

Holstein and Jersey cattle were immunized with a curli-producing strain of Escherichia coli (pCRL65/A012) or a noncurli-producing strain (pUC18/HB101) to determine differences in resistance to establishment of experimental intramammary infection. Cows (n = 6 per group) were immunized at 14 d prior to drying off, 7 d of involution, and at calving with 3 x 10(10) E. coli in Freund's Incomplete Adjuvant. At 30 d of lactation, one mammary quarter of each cow was infused with a wild strain of E. coli (727). Escherichia coli 727 was isolated from a naturally occurring intramammary infection and produced curli. All challenged quarters became infected, and all cows developed acute clinical mastitis. Geometric mean duration of intramammary infections was 6 d for both immunization groups. All infections were spontaneously eliminated within 10 d. No differences occurred between immunization groups in blood selenium and glutathione peroxidase activity, plasma selenium, number of E. coli 727 isolated from secretion after challenge, rectal temperature and SCC response, clinical status of mammary quarters, or DMI. Reduction in milk production after challenge was greater for cows immunized with E. coli pCRL65/A012. Immunization of dairy cattle with a curli-producing strain of E. coli did not protect against experimental intramammary challenge during lactation.     

Efficacy of intramammary immunization with an Escherichia coli J5 bacterin

Intramammary immunization was investigated as a procedure to reduce the clinical signs of coliform mastitis. Twenty-four cows were equally distributed to the following Escherichia coli J5 immunization schedules: 1) Subcutaneous injection 14 d prior to the end of lactation, intramammary immunization 7 d after drying off, and subcutaneous injection 30 d into the dry period; 2) subcutaneous injections at drying off, at 30 d into the dry period, and within 12 h after calving; and 3) unimmunized controls. Intramammary immunizations were the infusion of vaccine via the teat canal into each of the four mammary glands. Cows were challenged by infusion of E. coli 727 into one uninfected mammary quarter at approximately 30 d after calving. Intramammary immunization enhanced antibody titers against E. coli J5 and E. coli 727 compared with subcutaneous immunization. Immunoglobulin G titers against E. coli J5 and E. coli 727 in whey were greater at the time of challenge and 7 d after challenge for cows that received the intramammary immunization than for cows immunized by only subcutaneous injections. Serum IgG titers against E. coli 727 were enhanced at 7 d after challenge for cows receiving intramammary immunizations compared with conventionally immunized cows. Serum IgM titers against E. coli 727 were higher at calving for cows receiving intramammary immunization compared with conventionally immunized cows. Immunization schedule had minimal effect on systemic and local signs of clinical mastitis following challenge.     

Effects of adjuvants on safety and efficacy of an Escherichia coli J5 bacterin

The effects of using a water-soluble adjuvant or an emulsified oil-based adjuvant on the safety, antibody titer, and clinical responses of an Escherichia coli J5 bacterin were tested in an experimental infection trial. Fifty-one cows were assigned to 17 blocks of 3. Two cows within each block of 3 were vaccinated with a commercially prepared E. coli J5 bacterin containing either a water-soluble adjuvant or the same bacterin preparation emulsified in oil. One cow in each block was an unvaccinated control. Cows were immunized at drying off and 42 d later. The right or left front mammary quarter of each experimental cow was challenged by intramammary infusion of E. coli 727 between 14 and 35 DIM. Areas of inflammation at the primary injection site were greater 1, 2, and 3 d following primary vaccination for bacterin containing oil-in-water adjuvant compared with bacterin containing water-soluble adjuvant. Whey anti-E. coli J5 IgG titers were higher at calving for cows vaccinated with bacterin containing oil-in-water adjuvant than for cows either vaccinated with bacterin containing water-soluble adjuvant or unvaccinated controls. Serum x-E. coli J5 IgG titers were higher at calving for vaccinated cows than for unvaccinated controls. Peak bacterial counts in milk from challenged quarters were greater for unvaccinated controls than for cows vaccinated with bacterin containing water-in-oil adjuvant. Bacterial counts in milk from challenged quarters and clinical score both were greater in unvaccinated controls than cows vaccinated with bacterin containing water-in-oil adjuvant between 12 and 24 h postchallenge. Clinical responses were similar between unvaccinated controls and cows vaccinated with bacterin containing water-soluble adjuvant.     

Growth responses of coliform bacteria to purified immunoglobulin G from cows immunized with ferric enterobactin receptor FepA

The ability of purified bovine immunoglobulin (Ig) G from cows immunized with ferric enterobactin receptor FepA to inhibit the growth of coliform bacteria derived from bovine intramammary infection was investigated in iron-restricted media. All isolates of Escherichia coli (n = 21) and Klebsiella pneumoniae (n = 21) were tested for growth in a chemically defined medium containing 0.5 mg/ml of apolactoferrin and in a pooled source of dry cow secretion. The addition of 4 mg/ml of purified bovine IgG directed against FepA in the synthetic medium resulted in significant growth inhibition for both E. coli and K. pneumoniae isolates. Growth reduction of E. coli was greater than that of K. pneumoniae. In dry cow secretions, the growth of each E. coli isolate but of less than half of K. pneumoniae isolates (43%) was inhibited by IgG from cows immunized with FepA. Purified bovine IgG from cows immunized with E. coli J5 had a minimal inhibitory effect on the growth of both E. coli and K. pneumoniae isolates in the synthetic medium. In dry cow secretions, IgG from cows immunized with E. coli and K. pneumoniae isolates. Supplementation with 50 microM of ferric chloride to the medium completely reversed the inhibitory effects of the antibodies and lactoferrin. Bovine IgG directed against FepA apparently inhibited the growth of coliform bacteria by interfering with the binding of the ferric enterobactin complex to the cell surface receptor FepA.     

Anti-idiotypic responses of lactating cows immunized with monoclonal antibodies against bovine somatotropin.

The objective of this study was to produce anti-idiotypic antibodies with bovine somatotropin (bST)-like activity by active immunization of lactating cows and to determine their effects on milk yield. Several monoclonal antibodies against bST were evaluated for their interaction with bST in a rat growth bioassay. Two bST-agonist monoclonal antibodies (1 and 2), and two bST-antagonist monoclonal antibodies (3 and 4) were selected. Cows were immunized with immunoglobulin G as a control (n = 12) or with one of the four anti-bST monoclonal antibodies (1, 2, 3, 4; n = 12) on d 3, 24, 45, 66, 87, 108, 129, and 150 of lactation. From wk 3 of lactation, all cows immunized with each of the four anti-bST monoclonal antibodies developed anti-idiotypes until wk 30 of lactation. Total lactation yields were not different among monoclonal antibodies 2, 3, and 4 and control cows (9299, 9321, 9733, and 9415 kg, respectively). However, cows immunized with anti-bST monoclonal antibody 1 had reduced lactation yield compared with cows on other treatments (8136 kg). Daily milk yield of cows immunized with monoclonal antibody 1 was decreased from wk 9 of lactation [36.2 vs. 40.9 kg/d (control)] until the end of lactation, concomitantly with decreased bST concentration from wk 9 of lactation. Cows immunized with anti-bST monoclonal antibody 4 had increased milk yield compared with that of controls during wk 3 to 6 and wk 18 to 21 of lactation. Therefore, anti-idiotypes directed against anti-bST 1 had bST-antagonistic effects on lactation performance; anti-idiotypes against anti-bST 4 transiently increased milk yield.