紫外线—氯化锂复合诱变选育泰妙菌素高产菌株

目的 筛选泰妙菌素高产菌株。方法 以TMⅡ09—003为出发菌株,经UV照射180s后,将菌悬液均匀的涂布于加有0.8%的氯化锂平板上,在避光条件下于温度26℃～27℃,湿度60%～65%的环境下培养成熟后,进行挑取单菌落、初筛、复筛。以HPLC法测化学效价。结果 经UV照射180s后,氯化锂加量为0.8%剂量对菌株的致死率为88.6%。对用此剂量处理获得的诱变菌株进行了筛选,最终获得4支高产菌株,分别为：TMⅡ09—236、TMⅡ09—253、TMⅡ09—441和TMⅡ09—447。     

具有万古霉素高产能的东方拟无枝酸菌诱变株的选育研究

为提高万古霉素(Vancomycin)发酵生产水平,选育出其发酵生产的优良菌株,以东方拟无枝酸菌(Amycolatopsis oriertalsis)菌株V1806作为原始菌株,利用EMS(甲基磺酸乙酯)、UV(紫外诱变)和ARTP(常压室温等离子体诱变)等单一诱变、复合诱变的方法,选育出1株遗传稳定性好、且对万古霉素耐受性强的菌株Vua-15,其中试发酵效价达到7863 mg/L,比原始菌株提高了74.3%。研究结果表明,复合诱变对东方拟无枝酸菌菌种选育的效果优于单一诱变,更易选育出符合预期的优良菌株。研究不仅对万古霉素工业化发酵生产具有重要意义,而且为其他产品发酵菌种的选育提供了一定借鉴。     

不同诱变方法对TL-15028菌株产泰乐菌素相关性能的影响

为了选育出泰乐菌素高产菌株,以弗氏链霉菌(Streptomyces fradiae)TL-15028菌株作为出发菌株,利用化学诱变、物理诱变以及复合诱变的方法对菌株进行诱变处理,比较不同诱变方法的诱变效果,通过诱变筛选出1株遗传稳定性好、且具有泰乐菌素和豆油抗性的菌株TLEU-1503,其发酵效价比出发菌株提高了36.5%,达到14124μg/m L,对泰乐菌素高效生产、品质提升具有重要意义。     

紫外-氯化锂复合诱变选育泰妙菌素高产菌株

对泰妙菌素生产菌TMⅡ09-003经紫外线（UV）照射180 s后,将菌悬液均匀地涂布于加有0.8%的氯化锂平板上,在避光条件下于温度26℃～27℃、相对湿度60%～65%的环境下培养13～14 d后,挑取单菌落,进行初筛、复筛,用HPLC法测定含量。结果表明,UV照射180 s后,氯化锂加量为0.8%剂量对菌株的致死率为88.6%。对该剂量处理获得的诱变菌株进行了筛选,最终获得4支高产菌株,分别为TMⅡ09-236、TMⅡ09-253、TMⅡ09-441和TMⅡ09-447。     

不同诱变方法对TL-15028菌株产泰乐菌素相关性能的影响

为了选育出泰乐菌素高产菌株,以弗氏链霉菌(Streptomyces fradiae)TL-15028菌株作为出发菌株,利用化学诱变、物理诱变以及复合诱变的方法对菌株进行诱变处理,比较不同诱变方法的诱变效果,通过诱变筛选出1株遗传稳定性好、且具有泰乐菌素和豆油抗性的菌株TLEU-1503,其发酵效价比出发菌株提高了36.5%,达到14124μg/m L,对泰乐菌素高效生产、品质提升具有重要意义。     

诱变选育高产木聚糖酶Nystatin抗性菌株的研究

<正>自然界中植物性原料主要由纤维素、半纤维素和木质素3种成分组成,其中半纤维素(主要是木聚糖)的含量可达30%～35%,其在禾本科植物中含量最高。木聚糖是由木糖分子以β-1,4糖苷键连接而成的多聚体,外连阿拉伯糖和葡萄糖醛酸构成的侧链基团。木聚糖酶是专一降解木聚糖的一种复合酶,此外还有一些脱支链酶[1]。由于木聚糖在植物性原料中大量存在,使得木聚糖酶的应用非常广泛,如可用于纸浆的制造和漂白,降低含氯化合物用量,减少环境污染;在饲料上可以通过添加木聚糖酶来提高饲料的能量值和畜禽对     

螺旋霉素高产菌株的选育

以螺旋霉素链霉菌(Streptomyces spiramyceticus)SPM-14078菌株作为出发菌株,利用常压室温等离子体(ARTP)诱变、紫外线(UV)诱变和亚硝基胍(NTG)诱变的方法对菌株进行诱变处理,比较3种方法的诱变效果,通过诱变筛选出1株遗传稳定性好且具有Co~(2+)和豆油耐受性、噬菌体抗性的菌株SPMAR-1603,其发酵效价比出发菌株提高了294.9%,此结果对螺旋霉素高效生产、品质提升具有重要意义。     

紫外诱变选育黄霉素高产菌

采用紫外线诱变方法对黄霉素产生菌进行诱变处理,利用菌株抗自身代谢物的特征,在添加高浓度黄霉素的分离培养基上选育获得比出发菌株高的突变株sg2-U-5,效价比原始菌株提高了5.4倍。而且突变株的菌落形态与亲本菌株差异较大。     

中性蛋白酶高产菌株的选育

试验采用米曲霉(Aspergillus oryzae)菌株作为出发菌株,希望通过诱变处理,选育高产菌株,以达到工业化生产的需要。通过筛选和产酶试验,选育出的菌株P10,产中性蛋白酶酶活可达2000U/g,,比出发菌株提高了67%,产酶性能稳定,能够适应规模化生产的需要,实现了试验的预期目标。1材料和方法1.1材料1.1.1菌株现有的米曲霉菌株以及经诱变后纯培养得到的53个菌株,编号为A1-A28及P1-P25。1.1.2主要仪器与设备分析天平、恒温水浴锅、分光光度计、温度计、秒表、高压灭菌锅、超净工作台等。1.1.3主要试剂和溶液福林试剂、0.4mol/LNa2CO3、0.4mol/LC…     

亚硝酸诱变选育黄霉素高产菌的研究

采用亚硝酸诱变方法对黄霉素产生菌进行诱变处理,利用菌株抗自身代谢物的特征,在添加高浓度黄霉素的分离培养基上选育获得优良高产突变株sg2-N2-2,比发酵菌株的发酵单位提高了7.2倍,而且变株的菌落形态与亲本菌株差异较大。     

Cobalt metabolism in horse. Serum level and biosynthesis of vitamin B12.

The levels of serum vitamin B₁₂ were determined on 16 mature partly warm-blooded, partly Finnish rural-race horses by the radioisotopic competitive inhibition assay method. The mean value from three samplings carried out in dupli- or triplicate was 1.54 ± 0.16 ng/ml. The utilization of serum inorganic cobalt for cyanocobalamin synthesis was studied on two geldings, which received a dose of 200 µCi ⁵⁸CoGl₂ i.v. A Sephadex G-100 gel filtration was carried out with the serum proteins from serial blood samplings at different time intervals 15 min. to 48 hrs. after administration. The gel filtration showed the presence of two labelled proteins in the serum, one of them appearing some time after administration and disappearing almost completely towards the end of the experimental period. The two elution peaks are considered to represent inorganic ⁵⁸Co and ⁵⁸Co labelled vitamin B₁₂. The appearance of labelling in serum vitamin B₁₂ indicates the passing of cobalt into the intestine, and reabsorption into blood in the form of vitamin B₁₂.     

Reduction of the prevalence of footrot on New Zealand farms by vaccination

AIM: To compare serum analyses of vitamin B₁₂ and methylmalonic acid (MMA) as indices of cobalt/vitamin B₁₂ deficiency in lambs around weaning.

METHODS: Lambs on five properties, considered to be cobalt- deficient, were supplemented with either cobalt bullets, or short- or long-acting vitamin B₁₂ preparations. Blood samples, and in some cases liver biopsies, and liveweights were obtained at monthly intervals. Serum samples were assayed for vitamin B₁₂ and MMA and liver for vitamin B₁₂ concentrations. Pasture cobalt concentrations were measured on three of the properties.

RESULTS: Pasture cobalt concentrations were generally maintained below 0.07 μg/g dry matter (DM) on the properties sampled. Growth responses to supplementation were observed on only 2/5 properties, despite serum vitamin B12 concentrations being within the currently used ’marginal‘ reference range (336–499 pmol/L) for at least 3 months on all properties and in the deficient reference range (0–335 pmol/L) for at least 2 months on all farms except one. Serum MMA concentrations in supplemented lambs were <2 μmol/L, except in those animals sampled 1 month after receiving treatment with a short-acting vitamin B12 injection. Serum MMA concentrations in unsupplemented animals on properties on which no growth response to supplementation occurred generally reached peak levels of between 4 and 7 μmol/L at the nadir of serum vitamin B12 concentration. When a growth response was observed, differences in weight gain between supplemented and unsupplemented lambs occurred as mean serum MMA concentrations increased from 9 to 14 μmol/L. On one property where supplementation commenced before weaning, normal growth rates were maintained despite serum vitamin B12 concentrations of 140 pmol/L and serum MMA concentrations in excess of 40 μmol/L serum.

CONCLUSIONS: The possibility that current serum vitamin B₁₂ references ranges for diagnosis of cobalt deficiency are set too high and lead to over-diagnosis of responsiveness to cobalt/ vitamin B₁₂ supplementation is discussed. The suggestion is made that serum MMA concentrations in excess of 9–14 μmol/L will provide a more reliable diagnostic test for cobalt deficiency. However, there was sufficient variation between properties in the relationships between cobalt concentrations of pasture and serum vitamin B₁₂ or MMA concentrations to require more rigorous testing of the reliability of using serum MMA concentration for this purpose. The possibility that differences in rumen fermentation and therefore propionate and vitamin B₁₂ production could be involved is discussed. The measurement of serum MMA and vitamin B₁₂ appears to be of little value whilst the lamb is still suckling.

CLINICAL SIGNIFICANCE: Serum MMA concentration may offer advantages over serum vitamin B₁₂ concentrations in the diagnosis of a cobalt/vitamin B₁₂ responsiveness in weaned lambs.     

Vitamin B12 in the blood and in the liver of normal and of totally gastrectomized pigs

The concentration of vitamin R₁₂ in the blood was studied in five totally gastrectomized pigs followed for 10–18 months, and in four controls followed for 5¼–8 months. The concentration of vitamin B12 in the liver was investigated in three pigs of each group. The content of vitamin B₁₂ in the blood was of the same magnitude in both groups. Thus, removal of the stomach does not compromise the normal presence of this vitamin in the blood. The B₁₂ content in the liver was also unaffected by the operation, whereas it was increased after simultaneous administration of vitamin B₆.     

A critical evaluation of serum methylmalonic acid and vitamin B12 for the assessment of cobalt deficiency of growing lambs in New Zealand

AIM: To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B₁₂-responsiveness in lambs and critique existing serum vitamin B₁₂ reference ranges.

METHODS: Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B₁₂ and MMA, and liveweight gain.

RESULTS: Serum MMA concentrations were rarely elevated above the norm of <2 µmol/L when serum vitamin B₁₂ concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 µmol/L) unless serum vitamin B₁₂ concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 µmol/L).

CONCLUSIONS: The current reference ranges used in New Zealand for serum vitamin B₁₂ are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B₁₂ concentrations. Serum MMA concentrations <13 µmol/L indicate responsiveness to supplementation whilst concentrations <7 µmol/L indicate unresponsiveness. In the range 7–13 µmol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable.

CLINICAL RELEVANCE: The current reference ranges for vitamin B₁₂ responsiveness are conservatively high and lead to over-diagnosis of vitamin B₁₂ deficiency in ill-thriftiness of sheep.     

滤纸保藏法在泰乐菌素生产中的应用

通过对传统滤纸管保藏法的改良,应用于泰乐菌素生产菌的保藏,分别在不同的保藏周期时活化进行生产能力验证,结果显示改良滤纸管保藏法使用方便,摇瓶效价不低于沙土管保藏法,而且批间差异小,生产能力不随保藏时间的延长而降低。     

Growth response to increasing doses of microencapsulated vitamin B12 and related changes in tissue vitamin B12 concentrations in cobalt-de cient lambs

AIM: To investigate growth responses of cobalt-deficient lambs to increasing doses of microencapsulated vitamin B₁₂, and to measure associated changes in serum and liver vitamin B₁₂ concentrations over 243 days.

METHODS: From a flock grazing pastures that had low cobalt (Co) levels (about 0.06 mg Co/kg dry matter), 4-6-week-old lambs (n=137) were assigned to four groups and received either no treatment or a subcutaneous injection of 3.0, 4.5 or 6.0 mg of microencapsulated vitamin B₁₂ on Day 1. At approximately monthly intervals, all lambs were weighed and blood samples were collected from a selection (n=10) of monitor animals, up to Day 243. Liver biopsies were also carried out on the monitor lambs (n=8) on Days 1, 124 and 215.

RESULTS: The vitamin B₁₂-treated lambs grew significantly faster (p<0.001) than untreated animals. Liveweights after 243 days were 28, 45, 45 and 47 kg for the untreated, 3.0, 4.5 and 6.0 mg vitamin B₁₂-treated lambs, respectively. Of the initial group of untreated lambs, 68% had to be removed before the end of the trial because of substantial weight loss, but none of the treated animals were similarly afflicted. Serum vitamin B₁₂ concentrations increased in all vitamin B₁₂-treated lambs, reaching a peak at Day 25, and those of the 4.5 and 6.0 mg vitamin B₁₂-treated lambs remained significantly higher (except at Day 124) than the untreated lambs to Day 187. However, at Day 124, but not Day 215, the liver vitamin B₁₂ concentrations of treated lambs were two to three times higher than those of controls.

CONCLUSIONS: The growth rates of Co-deficient lambs were markedly improved by injection of 3.0, 4.5 or 6.0 mg of microencapsulated vitamin B₁₂, and liveweights were maintained for at least 243 days. Serum vitamin B₁₂ concentrations were related to this growth response; concentrations of <220 pmol vitamin B₁₂/l were associated with a 95% probability that lambs were Co-deficient and would thus respond to Co/vitamin B₁₂ supplementation. Based on these data, the current New Zealand reference criteria for Co deficiency should be reviewed.

CLINICAL SIGNIFICANCE: An injection of 3 mg microencapsulated vitamin B₁₂2 given to lambs at tailing will treat Co deficiency and will increase and maintain liveweights in a flock for up to 8 months.     

Serum Vitamin B12 Levels in Omani Goats

Blood and faecal samples were studied from 200 Omani goats of both sexes and various age groups, within five different regions in Oman. Survey questionnaires were undertaken on each goat. The serum vitamin B12 values ranged from 105 to >2000 pg/ml, with a mean value of 723.1 pg/ml and a 95% confidence interval of 61-1507 pg/ml. Kids in the age group 1-3 months showed significantly lower levels of serum vitamin B12 than the older animals. Varying levels of serum vitamin B12 were found in different regions of Oman and on farms within the same region. Goats maintained on commercial feed showed higher levels of serum vitamin B12 than those maintained on pasture or fed on household leftover food, as commonly practised in Oman. Coccidial counts were highest in animals with lower levels of serum vitamin B12. The results of this study indicate that there is a wide range of serum vitamin B12 levels in Omani goats and suggests that younger animals, with lower levels of serum vitamin B12, might be more susceptible to vitamin B12 deficiencies than older animals.     

Development of a specific radioimmunoassay for vitamin B12 and its application in the diagnosis of cobalt deficiency in sheep

A radioimmunoassay (RIA) for vitamin B₁₂ is described. Antisera were raised in rabbits using a conjugate between 5-O-succinyl cyanocobalamin and chicken serum albumin. The antibody is largely specific for the (upper) face of the vitamin B₁₂ molecule. The RIA can detect vitamin B₁₂ down to concentrations of 37 pmol/L and correlated well (r=0.980, p<0.001) with a commercial radioassay. The use of this RIA, together with an assay for plasma methylmalonic acid, in the diagnosis of cobalt/vitamin B₁₂ deficiency in sheep is described.     

Predicting copper status in beef cattle using serum copper concentrations

Copper deficiency is common in pasture-fed cattle in New Zealand⁽¹⁾. In general, the diagnosis of copper deficiency in a herd of cattle is based on a combination of history, examination of animals, examination of the environment, chemical analysis of blood, liver or pasture, and treatment response trials. The laboratory diagnosis of copper deficiency is currently based on liver and either plasma or serum concentrations of copper. Ellison⁽²⁾ reviewed the copper reference range for cattle used by the animal health laboratories in New Zealand and concluded that there is strong agreement in the literature that serum copper concentrations greater than 7.9 𝛍mol/l and liver copper concentrations greater than 95 𝛍mol/kg are adequate for young cattle. Furthermore, it has been reported that if copper concentrations in the liver are greater than 150–200 𝛍mol/kg wet weight, there is a negligible increase in serum copper as liver concentrations increase further⁽²⁾, with individual animal variation accounting for the range of values in serum copper at this point (7.9–18 𝛍mol/l).     

The effect of low levels of dietary cobalt on the chemiluminescence response of polymorphonuclear leukocytes of goats

Twenty ten-week-old newly weaned male Batinah goats were randomly assigned to a control (n = 10) and a treated (n = 10) group and were fed a diet containing 0.1 mg/kg DM cobalt (Co). Goats in the treated group received bi-monthly subcutaneous injections of 2000 μg of hydroxycobalamin. The phagocytic function of the polymorphonuclear leukocytes (PMN) were tested using a luminol-dependent chemiluminescence assay with opsonized zymosan as the phagocytic target. One month after the onset of the experiment PMN from the control group exhibited a significantly (p < 0.05) lower CL response, which continued for the second month. The results of the present study demonstrated that low levels of dietary cobalt leads to an early impairment of phagocytic function. This may at least in part, be an explanation as to why at the field level in Oman young goats fed diets containing low levels of Co appear to be more susceptible to infections.