Sequence and phylogenetic analysis of the 16S rRNA gene of Ehrlichia canis strains in dogs with clinical monocytic ehrlichiosis

The purpose of this study was to characterize, at the molecular level, the Ehrlichia canis strains involved in naturally occurring canine monocytic ehrlichiosis (CME) in Greece, and to investigate if any sequence diversity exists between the 16S rRNA genes of those involved in the mild non-myelosuppressive or the severe myelosuppressive form of CME. To this end, amplification of the ehrlichial 16S rRNA gene was attempted by nested polymerase chain reaction (PCR) assays in bone marrow (BM) aspirates from 20 dogs tentatively diagnosed as having non-myelosuppressive (n=10, group A) or myelosuppressive (n=10, group B) CME. PCR assay using E. canis-specific primers revealed that 15 BM samples, including all group A and 5 group B dogs, were positive. Using universal PCR primers, a nearly full-length 16S rRNA gene could be amplified from 13 BM samples, including 9 group A and 4 group B dogs. The 16S rDNA analysis based on secondary structure revealed that all sequences of the Greek strains were identical to each other and indicated 100% identity among some American (Venezuelan and Brazilian), European (Greek), Middle Eastern (Turkish) and Asiatic (Thailand) strains. The results of this study suggest that the E. canis strains involved in the non-myelosuppressive and myelosuppressive forms of CME in Greece share an identical 16S rRNA genotype.     

A proposal of clinical breakpoints for amoxicillin applicable to porcine respiratory tract pathogens

In the present position paper, an attempt was made to establish clinical breakpoints of amoxicillin to classify porcine respiratory tract pathogens as susceptible, intermediate or resistant based on their minimum inhibitory concentrations of amoxicillin. For this, a thorough review of the published literature with regard to swine-specific pharmacological data (including dosages of amoxicillin applied and routes of administration used), clinical efficacy, and in vitro susceptibility of the target pathogens was performed. Based on the comparative analysis of the results, the working group "Antibiotic Resistance" of the German Veterinary Medical Society (DVG) proposed to classify porcine respiratory tract pathogens that show MIC values of amoxicillin of < or =0.5microg/ml as "susceptible", those with MICs of 1microg/ml as "intermediate", and those with MICs of > or =2microg/ml as "resistant".     

Pain management in horses and farm animals

Objective: This review discusses the different analgesic drugs and routes of administration used in large animals for acute pain management. General guidelines and doses are given to assist in choosing techniques that provide effective analgesia. Etiology: Noxious stimuli are perceived, recognized, and localized by specialized sensory systems located at spinal and supraspinal levels. Diagnosis: Localizing the source of the noxious stimulus as well as understanding the behavioral aspects and physiological changes that result from such insult is important to adequately diagnose and treat pain. Pain assessment is far from being definite and objective; not only are there species differences, but also individual variation. In addition, the behavioral and physiological manifestations vary with the acute or chronic nature of pain. Therapy: Pain management should include (1) selecting drugs that better control the type of pain elicited by the insult; (2) selecting techniques of analgesic drug administration that act on pathways or anatomical locations where the nociceptive information is being processed or originating from; (3) combining analgesic drugs that act on different pain pathways; and (4) provide the best possible comfort for the animal. Prognosis: Providing pain relief improves the animal's well being and outcome; however, interpreting and diagnosing pain remains difficult. Continuing research in pain management will contribute to the evaluation of the pathophysiology of pain, pain assessment, and newer analgesic drugs and techniques.     

Variation among stallions in sperm quality after single layer centrifugation

Although single layer centrifugation (SLC) selects robust spermatozoa from stallion semen, the effect of individual variation has not been studied in detail. The objective of this study was to determine the variation among stallions in the effects of SLC on sperm quality during cooled storage for up to 48 hr. Semen samples from seven stallions (18 ejaculates) were split, with one portion being used for SLC and the other serving as a control (CON). Sperm quality (kinematics, reactive oxygen species (ROS) production, membrane integrity (MI) and chromatin integrity) were analysed at 0, 24 and 48 hr using computer-assisted sperm analysis and flow cytometry. Sperm quality was better in SLC than in CON at all timepoints, especially chromatin integrity and MI (p < .0001 for both), and some categories of ROS production (e.g. proportion of live hydrogen peroxide negative spermatozoa, p < .0001), but the degree of improvement varied among stallions and type of ROS (p < .05–p < .0001). Total and progressive motility were also better in SLC samples than in CON at 24 and 48 hr (p < .0001), although the effect on sperm kinematics varied. The interaction of treatment, time and stallion was not significant. In conclusion, sperm quality was better in SLC samples than in CON, although there was considerable individual variation among stallions. The improvement in sperm quality, particularly in chromatin integrity, was clearly beneficial, and therefore the use of this technique would be warranted for all stallion semen samples.     

Comparison of cardiac output determined by arterial pulse pressure waveform analysis method (FloTrac/Vigileo) versus lithium dilution method in anesthetized dogs

Objective – To compare the determination of cardiac output (CO) via arterial pulse pressure waveform analysis (FloTrac/Vigileo) versus lithium dilution method. Design – Prospective study. Setting – University teaching hospital. Animals – Six adult dogs. Interventions – Dogs were instrumented for CO determinations using lithium dilution (LiDCO) and FloTrac/Vigileo methods. Direct blood pressure, heart rate, arterial blood gases, and end‐tidal isoflurane (ETIso) and CO₂ concentrations were measured throughout the study while CO was manipulated with different depth of anesthesia and rapid administration of isotonic crystalloids at 60 mL/kg/h. Measurements and Main Results – Baseline CO measurements were obtained at 1.3% ETIso and were lowered by 3% ETIso. Measurements were obtained in duplicate or triplicate with LiDCO and averaged for comparison with corresponding values measured continuously with the FloTrac/Vigileo method. For 30 comparisons between methods, a mean bias of ?100 mL/kg/min and 95% limits of agreement between ?311 and +112 mL/kg/min (212 mL/kg/min) was determined. The mean (mL/kg/min) of the differences of LiDCO?Vigileo=62.0402+?0.8383 × Vigileo, and the correlation coefficient (r) between the 2 methods 0.70 for all CO determinations. The repeatability coefficients for the individual LiDCO and FloTrac/Vigileo methods were 187 and 400 mL/kg/min, respectively. Mean LiDCO and FloTrac/Vigileo values from all measurements were 145 ± 68 mL/kg/min (range, 64–354) and 244 ± 144 mL/kg/min (range, 89–624), respectively. The overall mean relative error was 48 ± 14%. Conclusion – The FloTrac/Vigileo overestimated CO values compared with LiDCO and the relative error was high, which makes this method unreliable for use in dogs.     

Effect of simulated acid rain on mycorrhizal infection of Pinus strobus L.

Mycorrhizal infection but not growth of white pine seedlings was reduced by application of simulated rain at pH 3.5 at 3 times ambient rates to plants grown in steamed Mardin soil inoculated with Pisolithus tinctorius. In unsteamed Mardin soil, the simulated acid rain at 3 times ambient rates had no effect on mycorrhizal infection or growth of pine seedlings if the rain was applied to the plants and soil or to only the soil before planting, except that nitrate-containing acid rain increased growth, infection and N content. In limed Mardin soil, the simulated acid rain reduced mycorrhizal infection if applied to the plants and soil as the seedlings were growing but not if applied to the soil before planting. Application of pH 3.5 rain at ambient rates had no detectable influence on mycorrhizal infection in 6 of 9 soils tested, but it stimulated infestion in 3 soils if the simulated rain contained only sulfate and in 1 soil if it had both sulfate and nitrate.     

Aerosol composition of urban and desert origin in the Eastern Mediterranean. I

Aerosol samples were collected in Tel-Aviv, Israel, during two different atmospheric situations, clear days and dust storms, and were analyzed for shape, size distribution, elemental composition, and presence of sulfate in individual particles. Results of two cases are presented in this paper. In both cases about 80% of the 0.2 to 2.0 µm particles contained sulfate. On the clear day over 50% of the total were pure sulfates while 27% were mixed sulfates; on the dusty day 63% were mixed sulfates — desert particles coated with sulfate — and only 20% were pure sulfates. The sulfate content of the desert particles was fairly high, on the order of 0.1 g sulfate g^?1 of desert particles. A possible explanation of this phenomenon may be heterogeneous nucleation of SO₂ on the surfaces of insoluble desert particles during their passage over the Mediterranean.     

Rapid detection of tetracyclines and their 4-epimer derivatives from poultry meat with bioluminescent biosensor bacteria

Tetracycline (TC) specific luminescent bacterial biosensors were used in a rapid TC residue assay sensitized to meet the EU maximum residue limit (MRL) for TC residues in poultry muscle tissue (100 microg kg(-1)) by membrane-permeabilizing and chelating agents polymyxin B and EDTA. Sensitivities of 5 ng g(-1) for doxycycline, 7.5 ng g(-1) for chlortetracycline, and 25 ng g(-1) for tetracycline and oxytetracycline were reached. Except for doxycycline, the MRLs of these tetracyclines include their 4-epimer metabolites. In the biosensor assay, all four 4-epimers showed induction capacity and antimicrobial activity, and antimicrobial activity was also observed in the inhibition assay, although with lower efficiency than that of the corresponding parent compound in both assays. The biosensor assay is an inexpensive and rapid high-throughput screening method for the detection of 4-epimer TC residues along with their parent compounds.