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.     

Multivariate statistical analysis of mass spectra as a tool for the classification of the main humic substances according to their structural and conformational features

The aim of this work is to explore the suitability of the complementary use of mass spectra and the corresponding statistical analysis (principal components-Pareto analysis (PCA) and discriminant analysis (DA)) of these spectra to differentiate diverse humic samples as a function of their structural and conformational features. To this end, the mass spectra of humic samples belonging to the main humic fraction types (gray humic acid, brown humic acid, and fulvic acid) were obtained by electrospray ionization mass spectrometry (ESI-MS). The results obtained showed that the application of PCA yielded a clear separation between blanks and humic samples. However, a clear differentiation among the humic fraction types was not achieved. The DA of PCA data, however, yielded a clear separation among the humic substances (HS) samples belonging to each HS fraction type considered: gray humic acids, brown humic acids, and fulvic acids. These results showed that the mass spectra of each humic sample include characteristic mass/charge (m/z) distribution values that can be considered as a "fingerprint" representative of its specific structural features. Our results also indicate that, although the m/z values principally corresponded to single-charged ions, we cannot identify these molecular weight distributions with those of humic samples, since sample molecular fragmentation, as well as partial molecular ionization, cannot be ruled out under our experimental and instrumental conditions.     

An additional life-history tactic of masu salmon: Migration of parr to coastal habitats

Alternative life-history tactics of masu salmon Oncorhynchus masou are well documented. Subsequent to the freshwater parr stage (age ≥ 1 + years), an anadromous form migrates to the sea after smolting, while a resident form matures without seaward migration. In addition to this typical migratory dimorphism, anecdotal reports based on field observations have indicated that some underyearling masu salmon use estuarine waters. However, no empirical evidence indicates saltwater utilisation and subsequent survival in the early parr stage. Here, we used otolith microchemistry to examine whether a portion of masu salmon parr in northern Japan enters coastal habitats. The otolith Sr:Ca ratios of most juveniles collected from six rivers had consistently low values, indicating that masu salmon parr inhabiting these rivers stay only in freshwater. In contrast, in individuals from a steep-gradient river the Sr:Ca ratios increased at about a 200-µm distance from the otolith core. These results suggest that some masu salmon parr might use brackish water or sea water temporarily. In addition, three masu salmon parr were found in another steep river where a culvert located only ten metres from the river mouth completely blocked upstream migration for spawning. The Sr:Ca ratios in these fish increased at about >200 µm from the otolith core, indicating the parr had immigrated to the non-natal river from the sea. Such flexible behaviour at an early life stage may contribute to the spatial expansion of masu salmon, and the movement could moreover help to stabilise its population dynamics.     

Effects of exercise on the morphology of the myenteric neurons of the duodenum of Wistar rats during the ageing process

This study aimed to evaluate the effects of regular physical activity on the morphology of the myenteric plexus of the duodenum in rats during the ageing process. To this end, 45 Wistar rats were divided into three groups: C (sedentary – 6 months old), S (sedentary – 12 months old) and T (trained – 12 months old). The animals of group S were given with a physical activity programme consisting of a 10-min-treadmill workout once a week. The animals of group T were submitted to the physical activity programme five times a week. Their duodenums were collected and submitted to the techniques of nicotinamide adenine dinucleotide (NADH)-diaphorase enzyme histochemistry for whole-mount preparations and transmission electron microscopy. No differences in the constitution of the myenteric plexuses were found when the sedentary and trained groups were compared with the control group. The ultrastructural features were similar for the three groups. However, it was verified that the physical activity of the trained animals resulted in a similar myenteric neuron morphology to that of the adult animals (6 months old), thereby confirming its beneficial effect, as the sedentary animals had larger alterations in the collagen fibrils and the basal membrane that occur through ageing. The quantitative analysis showed that the NADH-diaphorase positive neurons decreased with ageing and increased with physical activity ( P  > 0.05). No significant alteration ( P  > 0.05) in the neuronal profile area of the NADH-diaphorase positive neurons has been observed with ageing.     

Molecular cloning and sequence analysis of feline interferon-stimulated gene 15

The interferon-stimulated gene 15 (ISG15) is induced by type I interferon (IFN). Recent studies have revealed that like ubiquitin, ISG15 is conjugated with target proteins. In this study, the feline ISG15 (FeISG15) gene was cloned from feline IFNomega (FeIFNomega)-stimulated feline kidney epithelial (CRFK) cells. According to gene sequence results, cDNA was 474bp long and encoded a protein of 157 amino acids. The putative amino acid sequences showed 62.5-72.1% identity with those of other mammalian ISG15s. Similar to human and mouse ISG15, FeISG15 included tandem ubiquitin-like domains; its homology with feline ubiquitin was 36.3-39.5%. The LRLRGG conjugating motif was located only in the carboxyl terminal ubiquitin-like domain. FeISG15 also lacked the carboxyl terminal extension after the LRLRGG motif, which is present in mouse and human ISG15. Recombinant FeISG15 protein was expressed as a His-tagged fusion protein in Escherichia coli and purified by ion-exchange chromatography followed by affinity chromatography. Monoclonal anti-FeISG15 antibodies revealed free FeISG15 and FeISG15 conjugated with target proteins in cells after IFNomega stimulation by Western blotting analysis. Furthermore, mRNA of IFNgamma was detected from peripheral blood mononuclear cells (PBMCs) after stimulation with rFeISG15 extracellularly by RT-PCR. Taken together, these results suggested that FeISG15 had ubiquitin- and cytokine-like activity, as in other species.     

Pharmacokinetic disposition of triclabendazole in cattle and sheep; discrimination of the order and the rate of the absorption process of its active metabolite triclabendazole sulfoxide

A comparative pharmacokinetic study was conducted to determine the order and the rate of absorption of triclabendazole (TCBZ) in cattle and sheep. A commercial suspension of TCBZ (Biofasiolex, Biogénesis S.A., Argentina) was administered at a dose rate of 10 mg/kg by the oral route to six Holstein female calves and six Corriedale female sheep. The plasma concentration profiles of the metabolites triclabendazole sulfoxide (TCBZ-SO) and triclabendazole sulfone (TCBZ-SO₂) were analysed by means of the non-compartmental method. The order of the absorption process of the active metabolite, TCBZ-SO, was determined by construction of curves of cumulative absorbed fraction of the drug by means of the Wagner-Nelson method. The appearance of TCBZ-SO in plasma of cattle and sheep resembles the entry of a constant quantity of drug into the organism per unit time. This is explained by the reservoir effect of the rumen, which acts as a biological slow-release system for TCBZ-SO and its precursor TCBZ to the posterior digestive tract where they are absorbed. The plasma concentration profiles of TCBZ-SO in both species were well described by a one-compartment open model with zero-order process of absorption and first-order process of elimination. The values of AUC_0-∞ and C _max of TCBZ-SO did not differ between species, while other kinetic parameters except for λ _z had higher values in calves than in sheep. In the case of TCBZ-SO₂, t _max was the only parameter that did not differ between species, while other kinetic parameters except for λ _z had higher values in calves than in sheep.     

Assessment of brachial plexus blockade in chickens by an axillary approach

ObjectiveTo assess the brachial plexus block in chickens by an axillary approach and using a peripheral nerve stimulator.Study designProspective, randomized, double-blinded study.AnimalsSix, 84-week old, female chickens.MethodsMidazolam (1 mg kg⁻¹) and butorphanol (1 mg kg⁻¹) were administered into the pectoralis muscle. Fifteen minutes later, the birds were positioned in lateral recumbency and following palpation of the anatomic landmarks, a catheter was inserted using an axillary approach to the brachial plexus. Lidocaine or bupivacaine (1 mL kg⁻¹) was injected after plexus localization by the nerve stimulator. Sensory function was tested before and after blockade (carpus, radius/ulna, humerus and pectoralis muscle) in the blocked and unblocked wings. The latency to onset of motor and sensory block and the duration of sensory block were recorded. A Friedman nonparametric one-way repeated-measures anova was used to compare scores from baseline values over time and to compare the differences between wings at each time point.ResultsA total of 18 blocks were performed with a success rate of 66.6% (12/18). The latency for motor block was 2.8 ± 1.1 and 3.2 ± 0.4 minutes for lidocaine and bupivacaine, respectively. The latencies for and durations of the sensory block were 6.0 ± 2.5 and 64.0 ± 18.0 and 7.8 ± 5.8 and 91.6 ± 61.7 minutes for lidocaine and bupivacaine, respectively. There was no statistical difference between these times for lidocaine or bupivacaine. Sensory function was not abolished in nonblocked wings.Conclusions and clinical relevanceThe brachial plexus block was an easy technique to perform but had a high failure rate. It might be useful for providing anesthesia or postoperative analgesia of the wing in chickens and exotic avian species that have similar wing anatomy.     

Integrating animal movement with habitat suitability for estimating dynamic migratory connectivity

Context

High-resolution animal movement data are becoming increasingly available, yet having a multitude of empirical trajectories alone does not allow us to easily predict animal movement. To answer ecological and evolutionary questions at a population level, quantitative estimates of a species’ potential to link patches or populations are of importance.

Objectives

We introduce an approach that combines movement-informed simulated trajectories with an environment-informed estimate of the trajectories’ plausibility to derive connectivity. Using the example of bar-headed geese we estimated migratory connectivity at a landscape level throughout the annual cycle in their native range.

Methods

We used tracking data of bar-headed geese to develop a multi-state movement model and to estimate temporally explicit habitat suitability within the species’ range. We simulated migratory movements between range fragments, and calculated a measure we called route viability. The results are compared to expectations derived from published literature.

Results

Simulated migrations matched empirical trajectories in key characteristics such as stopover duration. The viability of the simulated trajectories was similar to that of the empirical trajectories. We found that, overall, the migratory connectivity was higher within the breeding than in wintering areas, corroborating previous findings for this species.

Conclusions

We show how empirical tracking data and environmental information can be fused for meaningful predictions of animal movements throughout the year and even outside the spatial range of the available data. Beyond predicting migratory connectivity, our framework will prove useful for modelling ecological processes facilitated by animal movement, such as seed dispersal or disease ecology.

     

Comparison of Climate Change Effects on Pome And Stone Fruit Phenology Between Balkan Countries and Bonn/Germany

The objective of the joint project between the two universities of Bonn and Sarajevo was to study the effects of climate change associated with warmer winter and spring temperatures on fruit tree phenology. Changes in flowering date of apple and cherry were correlated with local weather data obtained concomitantly on-site at four locations in the Balkan countries Slovenia, Serbia, B & H and at Bonn, Germany.Phenological data of flower opening (F1; BBCH 61) of apple cv. ‘Golden Delicious’ originated from 55 years at Cacak (44?°N, Serbia) and at Klein-Altendorf, University of Bonn (50?°N), cv. ‘Bobovec’ from the University of Ljubljana (46?°N, Slovenia) and cherry cv. ‘Burlat’ (35 years at Klein-Altendorf, 50?°N) and 20–30 years at Tuzla (44?°N, B & H) and Mostar (43?°N, Bosnia & Herzegovina; B & H).The annual precipitation showed a negligible change in the last 55 years of ±5% in the Balkan countries (e.?g. Cacak, 699?mm), but only ±1.8% at Klein-Altendorf (605?mm) with a slight shift to more winter rainfall.Regression analysis showed an increase in mean temperature of 1.7?°C (annual) and 1.45?°C for the vegetation period at Klein-Altendorf (last 55 years), Germany. In the evaluated Balkan stations, the greatest increase in annual air temperature between two climate phases (until 1987 versus from 1988 until now) was at Ljubljana (1.4?°C) and Cacak (1.2?°C) in contrast to the least increase (0.3?°C) at Tuzla (annual average 10?°C) and an intermediate value (0.9?°C) in the warmer Mostar (annual average 14.5?°C).This recent temperature increase advanced flower opening by 10–14 days for apple such as cv. ‘Golden Delicious’ at Cacak and 7–10 days (last 55 years) at Bonn and 4–5 days (last 35 years) for the early cherry cv. ‘Burlat’ at Klein-Altendorf/Bonn; at Mostar and Tuzla, the effect of climate change was greater with the early cherry cv. ‘Burlat’ (26 days) compared with the difference of the late cherry cultivar (16 days). Before recent climate change (1973–1987), cherry flower was advanced by 10 days in the warmer med climate at Mostar but delayed by 16 days in the cooler Tuzla. The results, i.?e. the flower advancement in the Continental climate but flower delay in the med climate, are discussed with respect to the risk of late frost and countermeasures to combat climate change effects.