鬼针草血清与抗菌药联合对耐药E.coli体外抑菌的研究

为了探讨鬼针草血清与抗菌药联合诱导细菌传代的体外抑菌活性,试验制备了鬼针草含药血清,采用二倍微量稀释法体外检测鬼针草血清与抗菌药联合诱导耐药大肠杆菌的抑菌传代情况。结果表明:鬼针草血清与抗菌药联合诱导细菌传代对耐药大肠杆菌有不同程度的抑菌活性。说明鬼针草血清可明显增强β-内酰胺类、氨基糖苷类和酰胺醇类抗菌药对耐药菌的抑制作用。     

救必应不同提取物与西药联合对产ESBLs细菌作用研究

试验旨在探讨4种救必应提取物与抗菌药物联合诱导细菌传代的体外抑菌活性。本试验制备了4种救必应提取物,采用96孔反应板二倍微量稀释法体外检测提取物与抗菌药物联合诱导耐药大肠杆菌传代的抑菌作用。结果显示,4种救必应提取物与抗菌药物联合诱导细菌传代对耐药大肠杆菌有不同程度的抑菌活性,其中提取物Ⅰ、Ⅲ与抗菌药物联合效果最好。提示,救必应提取物Ⅰ、Ⅲ与抗菌药物联合诱导细菌传代具有体外抗耐药菌作用。     

凉粉草含药血清与抗菌药联用对携带fosA3耐药基因的产CTX-M型ESBLs大肠埃希菌的抑菌效果

为了探讨凉粉草含药血清与抗菌药联合使用对连续传代的携带fos A3耐药基因的产CTX-M型ESBLs大肠埃希菌的体外抑菌活性,应用二倍微量稀释法测定各抗菌药及凉粉草含药血清对细菌的最小抑菌浓度(MIC)值,以凉粉草含药血清的亚抑菌浓度(1/2 MIC)与抗菌药联合使用对大肠埃希菌进行传代。结果发现,凉粉草含药血清联合抗菌药对大肠埃希菌有不同程度的抑菌活性,与凉粉草联合使用可显著增强氨基糖苷类、生物碱类、多磷类、利氟霉素类、磺胺类抗菌药对耐药菌的抑制作用。     

金不换水提取物与抗菌药联合对含NDM-1不动杆菌的体外作用研究

本试验旨在探讨金不换水提取物与抗菌药联合对含新德里金属β-内酰胺酶-1 (New Delhi metallo-β-lactamase-1,NDM-1)基因不动杆菌传代的体外抑菌活性,为金不换的综合利用提供参考依据。试验采用二倍微量稀释法体外分别检测金不换水提取物与抗菌药对受试菌的最小抑菌浓度(MIC),并以亚抑菌浓度(即1/2 MIC)的金不换水提取物与抗菌药联合作用对含NDM-1不动杆菌进行传代。结果显示,金不换水提取物的MIC为0.25 g/mL,以0.125 g/mL的金不换水提取物与头孢曲松钠、阿莫西林、头孢噻肟、痢菌净、磺胺间甲氧嘧啶、头孢他啶、头孢噻呋钠、美罗培南8种抗菌药联合作用传代后MIC明显降低。综上所述,金不换水提取物部分联合抗菌药诱导耐药细菌体外抗菌活性明显增强,对含NDM-1的不动杆菌具有一定的抑菌作用。     

穿心莲中药血清联合抗菌药对含fosA3大肠杆菌的作用研究

为了探讨穿心莲中药血清与抗菌药联合体外抗含耐磷霉素基因fos A3(Fosfomycin Resistance Gene)大肠杆菌的效果,对临床分离的细菌进行鉴定,确定细菌所含耐药基因种类,采用二倍微量稀释法分别体外检测穿心莲水提取物,中药血清与抗菌药物的最小抑菌浓度(MIC),并以1/2 MIC的穿心莲中药血清与抗菌药联合诱导含fos A3大肠杆菌的传代试验。结果显示,分离的菌株为含fos A3大肠杆菌;穿心莲水提物的MIC为1.0 g/m L,中药血清的MIC为1.0 m L/m L,以0.5 m L/m L的穿心莲中药血清与头孢曲松钠、阿莫西林、诺氟沙星、粘杆菌素、痢菌净、磺胺间甲氧嘧啶、阿米卡星、头孢他啶、头孢噻呋钠、氟苯尼考和磷霉素等11种抗菌药联合诱导传代后MIC显著降低。表明穿心莲中药血清联合抗菌药体外诱导含fos A3大肠杆菌,抗菌活性明显增强,并对含fos A3型大肠杆菌具有一定的耐药逆转作用。     

秦皮水提取物与部分西药对含fosA3大肠杆菌的联合作用

本试验初步探讨秦皮水提取物与抗菌药联合诱导含fosA3型耐药基因大肠杆菌传代的体外抑菌活性,为耐药大肠杆菌感染疾病提供治疗参考方案。采用二倍微量稀释法体外分别检测秦皮水提取物与抗菌药物的最小抑菌浓度(MIC),并以1/2 MIC的秦皮水提取物与抗菌药联合诱导含(fosfomycin resistance gene,FosA3)型大肠杆菌的传代试验。秦皮水提取物的MIC为1.0g/mL,以0.5g/mL的秦皮水提取物与头孢曲松钠、阿莫西林、诺氟沙星、粘杆菌素、痢菌净、磺胺间甲氧嘧啶、阿米卡星、头孢他啶、头孢噻呋钠、氟苯尼考和磷霉素11种抗菌药联合诱导传代后MIC显著降低。结果显示:秦皮水提取物联合抗菌药诱导耐药细菌体外抗菌活性明显增强,对含fosA3型大肠杆菌具有耐药逆转作用。     

中药“救必应”水提取物对产ESBLs大肠杆菌耐药性的影响

“救必应”为冬青科植物铁冬青的树皮或根皮水浸出物制备,具有广谱的抗菌作用.为研究中药“救必应”对产超广谱β-内酰胺酶(ESBLs)大肠杆菌耐药性的影响,本研究将产ESBLs的E.coli在亚抑菌浓度的中药“救必应”中传代,采用微量稀释法测定抗生素对产ESBLs的E.coli最小抑菌浓度(MIC),并通过透射电镜观察中药作用后细菌形态学变化.结果显示,中药“救必应”可以显著降低产ESBLs的E.coli对阿莫西林、加替沙星、诺氟沙星、盐酸左旋氧氟沙星、乙酰甲喹、磺胺间甲氧嘧啶钠、克林沙星、氟苯尼考的MIC值,并且MIC均降低4倍;此外,中药作用后可以引起细菌细胞壁的皱裂、干瘪、折叠、缩短等形态变化.     

中药野菊花提取物与抗菌药联合对产ESBLs大肠杆菌抑制效果研究

试验旨在探讨野菊花提取物与抗菌药联合作用对产超广谱β-内酰胺酶（extended spectrum β-lactamases, ESBLs）大肠杆菌的体外抑制效果,为野菊花的进一步开发利用提供理论依据。采用2倍微量稀释法和中西药联合诱导传代法测定野菊花提取物、抗菌药及联合作用后的最小抑菌浓度（minimal inhibitory concentration,MIC）。结果显示,野菊花水提物对该耐药大肠杆菌的MIC值为250 mg/mL,且在野菊花水提物亚抑菌浓度（1/2 MIC）作用下进行中西药联合传代,其与头孢曲松钠、头孢噻呋钠、头孢噻肟钠、氟苯尼考、痢菌净、阿米卡星、林可霉素7种抗菌药联合作用后,抗菌药的MIC值明显降低。表明野菊花水提物能够显著降低产ESBLs大肠杆菌对部分抗菌药的敏感性。     

迷迭香酸与抗菌药联合对含fosA3基因细菌抑菌效果的研究

试验旨在探讨迷迭香酸与抗菌药联合体外抗含fosA3耐药基因大肠杆菌的效果。本试验对临床分离的细菌进行鉴定,确定细菌所含耐药基因种类,用牛津杯法测定迷迭香酸的抑菌活性,平板涂布法测定迷迭香酸的杀菌活性,采用96孔反应板二倍微量稀释法分别检测迷迭香酸与抗菌药的最小抑菌浓度(MIC),再用微量棋盘稀释法测定迷迭香酸与抗菌药联合作用后的分级抑菌浓度指数(FICI)。结果显示,分离到含fosA3耐药基因大肠杆菌;迷迭香酸的MIC为640 μg/mL,迷迭香酸与环丙沙星、左旋氧氟沙星、黏杆菌素、加替沙星、阿米卡星、头孢他啶、头孢噻呋钠联合应用后其FICI≤0.5,有协同作用;与头孢曲松钠、诺氟沙星、痢菌净联合应用后0.5fosA3基因大肠杆菌有一定的抑菌活性和杀菌活性,其与部分抗菌药联合应用后,抗菌药体外抗菌活性显著增强。     

白头翁水提物与抗菌药联合对耐药大肠埃希菌作用研究

通过白头翁水提取物与抗菌药联合抗含耐磷霉素基因型和超广谱β-内酰胺酶型大肠埃希菌体外研究,为耐药菌感染的疾病防治提供理论依据。采用微量2倍稀释法测定白头翁水提取物与抗菌药的最小抑菌浓度(MIC),并用1/2 MIC的白头翁水提取物与抗菌药联合诱导耐药大肠埃希菌。结果表明,白头翁水提取物的MIC大于0.5g/mL,以0.5g/mL的白头翁水提取物与13种抗菌药联合使用,可增强抗菌药抗菌活性,显著降低抗菌药MIC。白头翁水提取物联合抗菌药可用于耐药细菌感染疾病的临床治疗。     

The effect of lemongrass oil and its major components on clinical isolate mastitis pathogens and their mechanisms of action on Staphylococcus aureus DMST 4745

The aims of this study were to investigate the antibacterial activity of lemongrass oil (LG) and its major components which were citral, geraniol and myrcene, against four strains of clinically isolated bovine mastitis pathogens, including Staphylococcus aureus, Streptococcus agalactiae, Bacillus cereus and Escherichia coli by the broth microdilution method, as well as their activity on S. aureus biofilm formation. Attempts to clarify their mechanisms of action by investigation of the effects on intracellular material leakage and morphological changes of S. aureus DMST 4745 were also made. The results demonstrate that S. agalactiae and B. cereus are more susceptible to LG, citral and geraniol than S. aureus and E. coli. Moreover, they also inhibit S. aureus biofilm formation and exhibit effective killing activities on preformed biofilms. The LG appears to have multiple targets in the bacterial cell, depending on concentration used as well as the amount of its components.     

Serum antibody coupled with the construction of gentamicin sulfate for the Escherichia coli targeted drug

Conjugates of the Escherichia coli serum antibody (Ab) with the gentamicin sulfate (GM sulfate) have been evaluated for the assessment of their ability to eradicate E. coli in vitro. The combination of every component in the targeted drug is the amino of serum antibody and gentamicin sulfate combined with the hydroxyl of PEG6000 by hydrogen bond, which forms stable complexes. The half-life of this complex is 4.83 h, and it is non-toxic side effects and low immunogenic. After the combination of antibody and gentamicin sulfate, the targeting of antibody is quite good. In vitro anti-bacterial activity of gentamicin sulfate increased 1000 times, and the clinical curative effect enhanced 100 times, this means higher efficacy and safety.     

Effect of bacitracin on tetracycline resistance in Escherichia coli and Salmonella

Tetracyclines and bacitracin are used extensively as a growth promotant and a therapeutic, respectively in livestock. In this study, we test whether there is an interaction between bacitracin and tetracycline that can select for an increase in microbial tetracycline sensitivity, using Escherichia coli and Salmonella as a model. We found via in vitro studies that bacitracin at sublethal concentrations preferentially selects for outgrowth of tetracycline-sensitive bacteria from among a population of tetracycline-resistant and tetracycline-sensitive E. coli and Salmonella strains. Most of the bacterial strains employed in our study were shown by PCR to possess the tetracycline resistance gene tet(A) or tet(C), either of whose activity is associated with tetracycline efflux. Bacitracin could be substituted for the lipophylic chelator, fusaric acid, used as a positive selective agent for tetracycline sensitivity. Together, these observations suggest that bacitracin-mediated selection for tetracycline sensitivity alters the function of tetracycline efflux proteins. Using bacitracin and tetracycline simultaneously may improve the effectiveness of the antibiotics, while decreasing the risk of selecting for a population of tetracycline-resistant microorganisms.     

Low levels of antibacterial drug resistance expressed by Gram-negative bacteria isolated from poultry carcasses in New Zealand

Abstract

AIM: To provide baseline data on the levels and patterns of antibacterial drug resistance expressed by Gram-negative bacteria isolated from poultry carcasses in New Zealand.

METHODS: Between July and December 2006, isolates of Escherichia coli (n=407) and Salmonella spp. (n=3) originating from carcass-rinse samples were submitted by testing laboratories affiliated to five major poultry processing plants. Isolates of Campylobacter jejuni (n=193) originating from retail poultry carcasses in 2005–2006 were retrieved from the Massey University archives. All isolates underwent disc diffusion susceptibility testing against panels of 12 (Enterobacteriaceae) and six (Campylobacter spp.) antibacterial drugs. Cephalothin-resistance in isolates of E. coli was confirmed using ETest strips, and confirmation of the resistance phenotypes for a subset of C. jejuni isolates used microbroth dilution assays. Patterns within the resistance phenotypes of the isolates were investigated using hierarchical clustering, and logistic regression modelling.

RESULTS: The majority of isolates (71.5% E. coli, 99% C. jejuni, and all three Salmonella spp. isolates) were fully susceptible to the drugs that were tested. Four (1%) E. coli isolates showed resistance to three or more drugs. The proportions of susceptible E. coli differed between the five processing plants. Resistances were detected in E. coli isolates, using disc diffusion to cephalothin (18.2%), ampicillin (4.4%), tetracycline (4.4%) and gentamicin (1.5%). There was an association between cephalothin-resistant isolates of E. coli and decreased susceptibility to gentamicin. Using ETests to ascertain the minimum inhibitory concentrations (MIC) of E. coli for cephalothin gave inconsistent results. One of 193 C. jejuni isolates was resistant to erythromycin, and microbroth dilution assays confirmed that this panel of C. jejuni was generally susceptible to antibacterial drugs.

CONCLUSIONS: The levels of resistance shown by Gram-negative bacteria isolated from chicken carcasses in New Zealand are among the lowest reported around the world. No resistance to extended-spectrum cephalosporin drugs was detected in E. coli, suggesting that CTX-M and AmpC beta-lactamases are rare or absent. Salmonella spp. are rarely isolated from poultry carcasses during routine testing in New Zealand, and the isolates identified during this study were fully susceptible to the drugs tested. A panel of C. jejuni isolates originating from retail poultry carcasses were susceptible to first-line and second-line antibacterial drugs. The use of cephalothin as a marker of resistance to first-generation cephalosporins may not be appropriate for non-type-specific E. coli of animal origin.     

Antibiotic-resistant Escherichia coli isolated from chilled meat at retail outlets

Antibiotic resistance was investigated in Escherichia coli isolated from beef, veal, lamb and pork at retail level. A total of 100 samples from each meat species was examined. About 16% of the 400 samples were contaminated with resistant E.coli. Significantly more E.coli isolates from pork were drug-resistant than isolates from other meats(P<0.01). About 7% of the combined beef, veal, and lamb E.coli isolates were resistant to two or more antibiotics compared to about 40.0% of the pork isolates (P<0.01) Transfer of resistance was observed for 39.2% of multiple resistant isolates. The results presented form a base for future monitoring of the presence of antibiotic-resistant coliforms on meat suitable for human consumption.     

Field trial evaluating changes in prevalence and patterns of antimicrobial resistance among Escherichia coli and Enterococcus spp. isolated from growing broilers medicated with enrofloxacin, apramycin and amoxicillin

The present study investigates, under field conditions, the influence of antimicrobial administration on prevalence and patterns of antimicrobial resistance among Escherichia coli and Enterococcus spp. isolated from growing broilers. For this purpose, a group of 16,000 commercial broiler chickens was treated with enrofloxacin from day 1 to day 3, gentamicin from day 19 to day 21, and ampicillin from day 26 to day 28. A control group of 16,000 broilers was placed in the same controlled environment poultry house. Fecal (from both groups) and feed samples were collected at regular intervals. Few E. coli isolates were obtained from either farm environment or poultry feed samples, while enterococci were found to be ubiquitous among these samples. The frequency of resistance against most antimicrobials tested was significantly higher (P < 0.05) in E. coli isolated from broilers receiving intermittent antimicrobial pressure than that from non-medicated broilers, whereas in enterococci these differences were only observed among structurally related antimicrobial drugs and over a short period of time. By the time the broilers reached market age (33 days), several multi-resistant E. coli and enterococci were detected in the feces of the medicated group. Results suggest that antimicrobial resistance in E. coli was mainly medication-dependent, whereas among enterococci, changes observed over time were apparently influenced by factors apart from antimicrobial exposure, namely the resistance organisms previously present in farm environment and those present in feedstuffs.     

In vitro reactivity and growth inhibition of EPEC serotype O111 and STEC serotypes O111 and O157 by homologous and heterologous chicken egg yolk antibody

IgY is a chicken egg yolk antibody which has been used for treatment and prophylaxis of gastrointestinal infections. Our aim was to verify if IgY obtained from chickens immunized with EPEC O111, STEC O111 and STEC O157 is able to show in vitro reactivity and biological activity towards the three bacteria. IgY was obtained from eggs laid before and after immunization with each bacterium. The preparations of IgY anti-EPEC O111 and anti-STEC O111 shared high reactivity detected by ELISA and growth inhibition ability towards both bacteria EPEC O111 and STEC O111. Nevertheless, the preparation of IgY anti-STEC O157 showed high reactivity and growth inhibitory effect only towards the homologous strain. Our results showing in vitro biological activity of IgY reinforce its use as an alternative for the treatment or prophylaxis of E. coli infections and encourage the development of in vivo studies for a possible future human therapeutic use.     

Performance and diarrhoea in piglets following weaning at seven weeks of age: Challenge with E. coli O 149 and effect of dietary factors

Four dietary factors (ad libitum versus feed restriction, control versus protein restriction at ad libitum feeding, control versus inclusion of lupin as a protein source at ad libitum feeding, and control versus extra vitamin E at ad libitum feeding) were tested in four separate experiments for the effect on diarrhoea. To introduce a diarrhoea-like condition, half of the piglets were challenged with an E. coli O 149 dose of 1 × 10⁸ colony forming units on days one and two after weaning (day of weaning = day zero). All piglets were susceptible since the dams were tested mono-zygotic susceptible to the attachment site of E. coli O 149 in the intestines. Each of the four experiments included 32 piglets from 4 sows. The design was a 2 × 2 factorial with dietary factor and E. coli O 149 challenge as the two factors, each at two levels. The piglets were housed individually during the experiment which lasted for 10 days from weaning at 7 weeks of age. The daily recordings included feed intake, weight and faecal score (from 1 = solid and cloddy to 6 = watery and yellow). Faeces from days 1 to 4 were tested for E. coli strains. In addition, blood was sampled and serum was analysed for antibodies to E. coli, IgG and IgM. Generally the E. coli challenge had no effect on growth and feed intake whereas faecal score and number of faecal haemolytic bacteria increased and faecal dry matter decreased. Feed restriction decreased the weight gain while faecal characteristics were unaffected. An analysis including all four experiments revealed that a feed intake of less than 200 g during the first day after weaning seems to be associated with a relatively high incidence of a post-weaning diarrhoea-like condition. Protein restriction decreased faecal score and increased faecal dry matter while weight gain tended to decrease. Inclusion of lupin affected neither weight gain nor faecal characteristics. Extra vitamin E did not affect weight gain while faecal dry matter decreased, and faecal score and number of faecal haemolytic bacteria increased. The dietary treatments had no effect on the measured immunoglobulins. In conclusion, the studied dietary factors could not alleviate a diarrhoea-like condition and at the same time maintain the growth rate. Furthermore, the results indicate that performance can be improved if piglets achieve a daily feed intake of at least 200 g during the first day after weaning.     

Resistance to Serum Complement,iss, and Virulence of Avian Escherichia coli

Control of avian colibacillosis is hampered by lack of easily identifiable markers for virulent Escherichia coli. Resistance to serum complement appears to be a widespread trait of virulent avian E. coli, suggesting that bacterial factors promoting survival in serum may be useful in discriminating between virulent and avirulent isolates. Such distinguishing factors may prove useful in diagnostic protocols or as targets in future colibacillosis control protocols. Interestingly, the factors responsible for resistance to complement differ in the E. coli isolated from mammalian and avian hosts, which may reflect differences in the nature of avian and mammalian colibacillosis. In some cases, genetic determinants for serum complement resistance in avian E. coli are found on aerobactin- or Colicin V-encoding plasmids. One such gene, iss, first described for its role in the serum resistance associated with a ColV plasmid from a human E. coli isolate, occurs much more frequently in isolates from birds with colibacillosis than in faecal isolates from healthy birds. Efforts to identify the genomic location of iss in a single, virulent avian E. coli isolate have revealed that it occurs in association with several purported virulence genes, all linked to a large conjugative R plasmid. At this time, it is not known whether iss merely marks the presence of a larger pathogenicity unit or is itself a contributor to virulence. Nevertheless, the presence of the complement-resistance determinant, iss, may be a marker of virulent avian E. coli exploitable in controlling avian colibacillosis.