Antimicrobial susceptibility testing of bacteria isolated from animals: considerations concerning the predefinition of breakpoints from the clinical pharmacological viewpoint

In vitro susceptibility tests are performed to receive information for selecting the most suitable antibacterial agent. As result of in vitro susceptibility tests, the minimum inhibitory concentration (MIC) indicates bacteria as resistant or sensitive. To determine MIC, therapeutically relevant breakpoints have to be defined. Microbiological criteria, chemical and physical characteristics as well as pharmacokinetic and toxicological (tolerance) properties of the antimicrobial compounds have to be considered in the selection of the therapeutic agent in addition to clinical experiences. Using some concentration and time dependent antibiotics as examples, it is demonstrated that the above mentioned criteria are not sufficiently considered in currently defined breakpoints.     

Results from a German interlaboratory test to establish the broth microdilution method for the determination of the minimum inhibitory concentration of bacteria from animals

In accordance with NCCLS guideline M31-A2, the DVG working group "antimicrobial resistance" developed a standard operating procedure (SOP) for the determination of the minimal inhibitory concentration (MIC) of antimicrobial agents by broth microdilution. This SOP was evaluated for its fitness for use in a national interlaboratory test. A total of 32 participating laboratories tested five strains (including two internationally accepted reference strains and three field strains representing in total three different bacterial species) three times at a one week interval each, using uniform microtitre plates. In 31 of the 32 laboratories more than 80% of MIC determinations performed yielded values in the expected range. In total 94.0% of the results were reproducible, with a lesser deviation of 4.0% from the expected values for laboratories performing MIC determination as a matter of routine (46.9%), compared to 7.9% for laboratories without such routine (53.1%). Comparing the consistency of results on the basis of the tested strains, a higher reproducibility of the results was observed for reference strains (96.1%) than for field strains (92.6%). In particular results obtained for the Streptococcus uberis field strain were afflicted with a higher error ratio (98 deviations from the expected values). Among the tested antimicrobial agents, a higher variability of results was recorded only for gentamicin with 16.7% divergent MIC determinations (mean value 6.0%). The high reproducibility of the results confirmed by this interlaboratory study underlines the robustness of the developed SOP as well as broth microdilutions as the method of choice for MIC determina tion.     

Cross-resistance between antimicrobial agents used in veterinary medicine: molecular background and practical consequences for susceptibility testing

Phenotypic resistance of veterinary pathogens to more than one antimicrobial agent (multi-resistance) may be caused by intrinsic resistance to the antimicrobial agents, acquired cross-resistance, or acquired co-resistance. Known cross-resistances allow to select so-called "representative substances" which are tested and the results of which can also be regarded as being valid for other members of the same class of antimicrobial agents. In general, a limitation in the number of antimicrobial agents to be tested in routine diagnostics is necessary because of capacity and cost efficiency. This is of particular relevance when the broth microdilution method - recommended as the method of choice - with 96-well microtiter plates is used. The knowledge about the relationship between different resistance phenotypes and the corresponding resistance mechanisms is of major value for both, the laboratory personnel and the veterinary practitioner. This review explains how "representative substances" for the most relevant classes of antimicrobial agents used in veterinary medicine are chosen on the basis of known cross-resistances.     

Remediation of salt-affected soil by natural and chemical amendments to improve berseem clover yield and nutritive quality

Salinity is a major problem affecting crop production worldwide. The potential of different soil amendments (sulphur, gypsum, compost, and combination of gypsum, compost and sulphur) and two plant-growth stimulators (potassium silicate and calcium carbonate nanoparticles) to alleviate the harmful effects of saline soil on two cultivars of berseem clover (Trifolium alexandrinum L.), one salt tolerant (‘Serw-1’) and the other high yielding (‘Helaly’), was evaluated. Some soil properties were measured before and after treatments. Plant growth parameters, herbage yield, nutritive value and plant metabolites, including photosynthetic pigment and endogenous hormone contents, were also determined. The herbage yield of berseem clover cultivars substantially increased with the application of soil amendments in both years. The greatest yield increase was obtained with the compost application. The highest protein contents in herbage (16.4% in Helaly and 17.2% in Serw-1) were detected in response to composite (compost + gypsum + sulphur) and compost treatments, respectively. Carbohydrate content increased to 9.53% in Helaly and 5.1% in Serw-1 in response to the gypsum and compost treatments, respectively. The composite, compost and gypsum soil amendments were the most effective treatments resulting in the greatest herbage yield and quality values.     

Isolation and molecular biological investigations of avian poxviruses from chickens, a turkey, and a pigeon in Croatia

In the last 3 yr, several outbreaks of avian poxviruses (APVs) have been observed in different parts of Croatia. Four strains of APVs, from chickens, a pigeon, and a turkey, were isolated from cutaneous lesions by inoculation onto the chorioallantoic membranes (CAM) of 12-day-old specific-pathogen-free chicken embryos. The resulting proliferative CAM lesions contained eosinophilic cytoplasmic inclusion bodies. The characteristic viral particles of poxvirus were detected in the infected CAM and also in the infected tissues by transmission electron microscopy. Further identification and differentiation of the four various APVs were carried out by the use of a polymerase chain reaction (PCR) combined with restriction enzyme analysis. Using one primer set, which framed a region within the APV 4b core protein gene, it was possible to detect APV-specific DNA from all four tested isolates. PCR results revealed no recognizable differences in size of amplified fragments between the different APVs from chickens, turkey, and pigeon. Restriction enzyme analysis of PCR products using NlaIII showed the same cleavage pattern for turkey and chicken isolates and a different one for the pigeon isolate. Multiplex PCR for direct detection of APV and reticuloendotheliosis virus (REV) was carried out to determine the possible integration of REV in the genome of isolated APVs. The obtained results revealed that REV was present in chicken and turkey strains of poxviruses, whereas the pigeon isolate was negative. It is not known whether the avipoxvirus vaccine strain used in Croatia is contaminated with REV or if the REV is naturally contaminating Croatian field strains of fowl poxvirus. The latter is indicated by the negative REV finding in the pigeon, which was not vaccinated. The results of the present study indicate the reemergence of fowlpox in Croatia, where infections have not been recorded since 1963 and never confirmed etiologically.     

Plasma chemistry reference values in hybrid falcons in relation to their species of origin

Twenty-one blood chemistry parameters were determined in 127 peregrine falcons (Falco peregrinus), 79 gyr-peregrine hybrids and 166 gyr-saker hybrids. These parameters, together with those previously established in 53 gyr falcons (Falco rusticolus) and 234 saker falcons (Falco cherrug), were compared. There were statistically significant differences in 15 of the parameters between the saker and peregrine falcons; the saker and gyr falcons; the saker and gyr-saker hybrids; the peregrine and gyr falcons; and the peregrine and gyr-peregrine hybrids, but not between the gyr falcons and the gyr-saker falcon hybrids.     

Effect of Nitrogen Fertilization and Foliar Application of Plant Growth Retardants and Zinc on Cottonseed, Protein and Oil Yields and Oil Properties of Cotton

Field experiments were conducted in two successive seasons at the Agricultural Research Centre, Giza, Egypt (on a clay loam soil), to determine the effects of N fertilization (added at rates of 107 and 161 kg N ha^–1) and foliar application of plant growth retardants (Pix, Cycocel and Alar; each applied once at 300 p.p.m., 75 days after planting) and zinc (applied at 0.0 and 50 p.p.m., two times, 80 and 95 days after planting) on cottonseed, protein and oil yields and oil properties of the Egyptian cotton cultivar Giza 75. The higher N rate, as well as the application of all growth retardants and zinc, resulted in an increase in cottonseed yield ha^–1, seed index, seed protein content, oil and protein yields ha^–1, seed oil refractive index, unsaponifiable matter and total unsaturated fatty acid content (oleic and linoleic). However, those treatments decreased the oil acid value, saponification value, and total saturated fatty acid content. The seed oil content tended to decrease when the high N rate was applied, but tended to increase with the application of all growth retardants and zinc. There were some differences between Pix, Cycocel and Alar regarding their effects on the studied characters. The highest increase in seed yield ha^–1, seed index, and oil and protein yields ha^–1 was found with Pix, followed by Cycocel. The Cycocel treatment gave the lowest total saturated fatty acid oil content, followed by Alar.     

Evaluation of a Mycoplasma gallisepticum strain exhibiting reduced virulence for prevention and control of poultry mycoplasmosis.

Two experiments were conducted to evaluate the virulence and vaccination efficacy of a Mycoplasma gallisepticum (MG) isolate designated MG Intervet 6/85. Virulence of the strain was determined by evaluation of airsacculitis scores following aerosol exposure to the isolate before and after 10 sequential passes in either commercial broiler chickens or commercial turkeys. Two-week-old specific-pathogen-free chickens were vaccinated by aerosol exposure. The birds were challenged with the R' strain of MG at either 4 or 8 weeks post-vaccination. Efficacy was evaluated by airsacculitis scores determined 21 days after challenge. Ten repetitive back-passes of the isolate in chickens and turkeys did not substantially increase the virulence. Virulence for both chickens and turkeys was minimal, while protection elicited by aerosol vaccination in young chickens against virulent R' strain was significant (P less than or equal to 0.05) compared with unvaccinated controls.     

家禽中新发生和重新出现的一些细菌性感染

细菌病仍然是有关经济和公共卫生的主要问题。细菌性疾病的发生往往伴随死亡率升高、增重减慢、产蛋下降和药物成本提高。近年来 ,由被污染禽类食品引起的公共卫生问题以及耐抗生素菌株的增多已成为一些国家公众所关注的主要的政治性话题。本文目的在于探讨一些引起全球养禽业问题的新近发生和重又出现的细菌性疾病。1　淀粉样关节病1 992年在荷兰的重型罗斯蛋种鸡和法国的一群罗斯蛋鸡中出现了一种从临床症状上还不被人们所认知的关节性疾病。其表现为生长减慢、全身性淀粉样病并发淀粉样关节病。通过实验室检测 ,从中分离到了粪肠球菌 (E…     

Safety assessment of Bt 176 maize in broiler nutrition: degradation of maize-DNA and its metabolic fate.

Insect resistant Bt 176 maize has been developed by genetic modification to resist European borer infection. In the present investigation, the experiment was conducted to determine the effect of feeding a new hybrid of Bt 176 maize (NX 6262- Bt 176) on general health condition and performance of broiler chickens. Maize grains and diets were subjected to proximate analysis. Amino and fatty acids investigation were applied for both maize grains before used. To evaluate the degradation of NX 6262- Bt 176 maize DNA and its metabolic fate in broiler blood, muscles and organs. One-day-old male broilers were fed ad libitum on either an experimental diet containing NX 6262- Bt 176 or a control diet containing the non-modified maize grains for 35 days. Feed consumption and body weight were recorded weekly during the experimental period. All chickens were subjected to nutritional evaluation period at day 20 of age for 5 successive days, to calculate the percentage of apparent digestible nutrients in both diets. At day 35 samples were collected at several intervals after feed withdrawal. Prior to slaughter blood samples were collected from all birds by heart puncture to prevent DNA cross contamination. Samples from pectoral and thigh muscles, liver, spleen, kidney, heart muscle, bursa and thymus glands were collected. Digesta from different sections of the gastrointestinal tract (GIT) were collected as well. Packed cell volume (PCV) and some serum parameters were investigated. There were no significant differences between control and experimental group concerning chemical composition of feeds, apparent digestible nutrients, and all performance parameters measured (P > 0.05). Furthermore, there were no differences in the PCV and the analysed serum parameters between the control and experimental group. The results of maize DNA digestibility showed that the new variety takes the normal physiological passage along broiler GIT similar to the conventional line. In addition, Bt 176 maize DNA appears to be partially degraded in different parts of GIT comparable to the DNA of the control maize line. Results of the metabolic fate of maize DNA in broiler blood, muscles and organs indicated that only short DNA fragments (199 bp) derived from the plant chloroplast gene could be detected in the blood, skeletal muscles, liver, spleen and kidney, which disappeared after prolongation the fasting time. In heart muscle, bursa of Fabricius and thymus, no plant chloroplast DNA was found. Bt gene specific constructs from Bt 176 maize were not detected in any investigated blood or tissue samples.