饲料中3种四环素类药物的高效毛细管电泳检测方法

作者建立了鸡饲料中四环素、土霉素和多西环素的高效毛细管电泳检测方法。3种药物在40 mmol/L磷酸氢二钠—20 mmol/L柠檬酸(pH 2.6)的电泳缓冲液、30 kV的分离电压、长度64.5 cm、内径75 μm的毛细管内得到完全分离。在1～80 mg/kg范围内3种药物线性良好,R2>0.994;配合饲料和预混料中3种药物的定量限分别为2和4 mg/kg;添加回收试验回收率>80%,方法的日内变异系数和日间变异系数均<10%。     

Delivering oxytetracycline to first-feeding zebrafish, Danio rerio (Hamilton), and goby, Asterropteryx semipunctata Rüppell, larvae using lipid spray beads

Lipid spray beads (LSB) were evaluated for delivery of the low-molecular weight, water-soluble antibiotic, oxytetracycline·HCl (OTC) to fish larvae. Various OTC core-to-lipid ratios and OTC core concentrations were evaluated to maximize OTC delivery efficiency by LSB. Acceptability and digestion of LSB containing OTC and riboflavin by larval zebrafish, Danio rerio (Hamilton), and larval gobies, Asterropteryx semipunctata Rüppell, were also evaluated. Increasing LSB core-to-lipid ratios from 1:3 to 1:1 v/v resulted in an increase in encapsulation efficiency (EE) from 2.33 to 3.68% w/w. Increasing OTC concentrations of core solutions from 0.1 to 0.5 g OTC mL⁻¹ increased EE from 3.95 to 18.77% w/w, respectively. Although retention efficiency (RE) was unaffected by this increase, delivery efficiency was increased to 7.9 ± 0.7% w/w, after correcting for leakage losses because of the suspension of beads in water for 60 min. Consumption of LSB containing OTC by first-feeding zebrafish and goby larvae was confirmed by analysis of feeding incidence and gut fullness. Visual observations of larvae fed on LSB containing riboflavin indicated that larvae of both species digested LSB. Zebrafish larvae fed on OTC LSB contained 39.3 ± 2.5 ng OTC after purging LSB from their guts. Use of LSB provides an effective means of delivering therapeutics to fish larvae and could greatly reduce required doses compared with current methods of immersing larvae in solutions of therapeutic agents.     

Comparison of lipid-walled microcapsules and lipid spray beads for the delivery of water-soluble, low-molecular-weight materials to aquatic animals

Microparticles (< 40 μm diameter) composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were used to encapsulate the low-molecular-weight (mol. wt 460) antibiotic oxytetracycline in the form of either oxytetracycline hydrochloride (OTC.HCl) or oxytetracycline hemicalcium salt (OTC.HEM). Dry, finely ground particles of core material were encapsulated in spray beads. Dissolved core material was encapsulated in lipid-walled microcapsules.
Oxytetracycline (OTC) was most efficiently delivered (≈ 46.5 mg g⁻¹ lipid after 24 h suspension in seawater) as a hemicalcium salt in spray beads. Lipid-walled microcapsules were most efficient for delivering OTC (≈ 8.7 mg g⁻¹ lipid) as OTC.HCl dissolved in 0.2  M HCl at a concentration of 300 mg mL⁻¹.
Spray beads containing OTC.HEM were very stable over 1 month in storage. Lipid-walled microcapsules containing aqueous OTC.HCl lost ≈ 30% of their core material during storage. Freeze-drying of both microparticle types did not improve storage of spray beads, but showed promise for reducing leakage from lipid-walled microcapsules during storage and delivery to suspension feeders.     

Antibiotics modulate biofilm formation in fish pathogenic isolates of atypical Aeromonas salmonicida

Atypical Aeromonas salmonicida causes furunculosis infections of non-salmonid fish, which requires antibiotic therapy. However, antibiotics may induce biofilm in some bacteria, which protects them against hostile conditions while allowing them to persist on surfaces, thus forming a reservoir for infection. The aim of this study was to determine whether atypical isolates of A. salmonicida increased biofilm in the presence of two antibiotics, florfenicol and oxytetracycline. A microtitre plate assay was used to quantify biofilm in the presence and absence of each antibiotic. Fifteen of 28 isolates formed biofilms under control conditions, while 23 of 28 isolates increased biofilm formation in the presence of at least one concentration of at least one antibiotic. For oxytetracycline, the most effective concentration causing biofilm to increase was one-quarter of that preventing visible bacterial growth, whereas for florfenicol it was one-half of this value. This is the first study to demonstrate that a bacterial pathogen of fish increases biofilm in response to antibiotics. Biofilm formation may increase the risk of re-infection in culture systems and this lifestyle favours the transmission of genetic material, which has implications for the dissemination of antibiotic-resistance genes and demonstrates the need for enhanced disease prevention measures against atypical A. salmonicida.     

三种有机溶媒制备土霉素注射液的稳定性试验

采用甘油甲缩醛、二甲基乙酰胺、聚乙烯吡咯烷酮K30三种有机溶媒为主,经特殊制备工艺制成长效土霉素注射液样品,在前期获得满意的刺激性试验与临床治疗效果基础上,进行了为期两年的常温条件下制剂含量、外观色泽、澄明度、pH值、鉴别反应等质量指标试验观察,为药品的生产制造提供理论依据。结果表明:不同有机溶媒配比样品的稳定性均好,符合进口兽药质量标准要求,但以甘油甲缩醛、二甲基乙酰胺和注射用水溶媒联合使用,制剂成本最低。     

土霉素在土壤中降解特性研究

采用室内培养法研究了不同温度条件下,土霉素在土霉素单一及土霉素-锌复合体系中的降解作用。结果表明,在4℃和25℃时,土霉素在单一及复合体系中的降解速率均随培养时间增加而降低,随土霉素添加浓度的增加而增加。与4℃相比,土霉素在25℃环境条件下降解速率显著提高,半衰期明显缩短。与单一污染相比,在中、高浓度锌胁迫条件下,土霉素的降解速率明显降低,半衰期明显延长。参考POPs国际公约关于化学品持久性的定义,土霉素在单一及复合体系中均属易降解有机物。     

碳氮比对鸡粪堆肥中土霉素降解和堆肥参数的影响

【目的】分析不同C/N处理下土霉素在鸡粪堆肥中的降解及其对堆肥理化性质的影响,以期阐明不同C/N处理对土霉素降解及堆肥腐熟度的影响。【方法】以鸡粪和小麦秸秆为原料,采用室内好氧堆肥法,研究鸡粪好氧堆肥过程中,不同C/N处理（T1：C/N=21.6、T2：C/N=25.5和T3：C/N=32.8）对鸡粪堆肥中土霉素的降解及堆体温度、pH、发芽指数等指标的影响。【结果】（1）C/N对鸡粪堆肥过程中土霉素的降解影响显著。整个堆肥期内,不同处理土霉素的降解速率大小顺序为T2＞T3＞T1。土霉素在各处理堆体中的降解均符合一级动力学方程,其相关系数介于0.9431—0.9967。（2）土霉素存在条件下,堆肥初始C/N对堆体温度是有影响的。C/N高的处理较C/N低的处理堆体温度上升速率快, 达到最高温所需的时间也较短,最高温也较高。（3）C/N对土霉素存在下鸡粪堆肥堆体可溶性氮含量影响显著。如堆肥结束时T1、T2和T3处理堆体NO3--N与堆肥初始相比分别升高78.50%、62.37%和59.34%。（4）高C/N处理在一定程度上可以显著降低堆肥浸提液的生物毒性。如堆肥结束时C/N为25.5和32.8的处理GI＞80%。【结论】对含有土霉素的鸡粪进行堆肥,较高的初始C/N（25.5-32.8）能够有效地促进鸡粪中土霉素的降解和鸡粪的腐熟。     

Antibacterial activity of oxytetracycline on microbial ecology of Nile tilapia (Oreochromis niloticus) gastrointestinal tract under laboratory condition

The antibacterial effect of oxytetracycline (OTC) on bacterial load, pathogen and possible development of antibiotic resistance in intestinal flora of apparently healthy Nile tilapia (Oreochromis niloticus) was investigated under laboratory condition. OTC, a broad spectrum approved antibiotic widely used in aquaculture, was fed to Nile tilapia through medicated diet at a rate of 2 g/ kg of feed for consecutive 7 days in treatment 1 (T₁) and 30 days in treatment 2 (T₂). Changes in physico‐chemical parameters of water were also recorded where pH and dissolved oxygen influenced bacterial load and coliform count respectively. At the start of the experiment, initial bacterial load in fish intestine was 1.61 ± 2.25 × 10¹⁰ cfu/g, which after OTC treatment decreased significantly (p < .05) to 3.06 ± 2.08 × 10⁸ cfu/g on day 7 and 3.45 ± 4.46 × 10⁷ cfu/g on day 30 when compared with the control group. Among the 10 bacterial genera identified in the intestine, 4 potential public health concern bacteria viz., Salmonella, Escherichia, Enterobacter and Staphylococcaus were predominant. There was a sharp increase in bacterial load on day 18 in T₂ that may be attributed to the development of antibiotic resistance in intestinal bacteria. Antibiotic susceptibility test for isolates against seven antibiotics: amoxycillin, ciprofloxacin, co‐trimoxazole, erythromycin, gentamicin, nitrofurantoin and tetracycline using disc diffusion method revealed significantly increased resistance of Gram‐negative rods (p < .05) that possibly caused increased frequency of OTC‐resistant microorganisms. In conclusion, short‐ and long‐term exposure to OTC treatment affected the distribution of bacterial genera including pathogens in the Nile tilapia gastrointestinal tract and concomitantly influenced their antimicrobial resistance.     

Feed intake and growth of Arctic charr, Salvelinus alpinus (L.), fed diets supplemented with oxytetracycline and squid extract

This study examined the long-term effects of a potential feeding deterrent, oxytetracycline (OTC), and a putative feeding stimulant, squid extract, on feed intake, growth and dry matter (DM) digestibility in Arctic charr, Salvelinus alpinus (L.). Fish were fed one of four diets for 9 weeks: 1. commercial feed formulation (basic); 2. BM (basic plus 20 g kg⁻¹ OTC); 3. BMS (BM plus 10 g kg⁻¹ squid extract); 4. BS (basic plus 10 g kg⁻¹ squid extract). The feed intake of Arctic charr was not reduced by the addition of OTC to the basic feed, and addition of squid extract to the medicated feed did not enhance feed consumption. DM digestibility and growth tended to be reduced in groups of fish fed diets supplemented with the antibiotic (BM and BMS). This suggests that OTC may have interfered with nutrient digestion and absorption. OTC does not seem to be a feeding deterrent for Arctic charr, possibly indicating that the charr is a rather unselective feeder. Because the medicated feeds were readily accepted, there may be little to be gained by addition of feeding stimulants to these types of feeds for charr.     

冬枣溃疡病药剂防治研究

冬枣溃疡病是为害冬枣枣吊的一种细菌性病害,用不同药剂对其开展了室内毒力测定和大田药效试验。结果表明,室内抑菌效果比较好的是72％农用链霉素和88％水合霉素,大田防治效果最好的是72％农用链霉素,防治效果为66．34％。