Determination of beta-lactams in milk using a surface plasmon resonance-based biosensor

Two surface plasmon resonance (SPR)-based biosensor assays for detection of beta-lactam antibiotics in milk are reported. The assays are based on the enzymatic activity of a carboxypeptidase converting a 3-peptide into a 2-peptide, a reaction that is inhibited in the presence of beta-lactams. Antibodies were used to measure either the amount of formed enzymatic product or the amount of remaining enzymatic substrate. Both assays detected different beta-lactams at or below European maximum residue limits (MRLs), and the detection limit for penicillin G was 1.2 microg/kg and 1.5 microg/kg for the 2- and 3-peptide assays, respectively. The precision (CV) was < 5%, both within and between assays at the penicillin G MRL (4 microg/kg). The biosensor results obtained upon analysis of incurred milk samples were compared with results obtained by liquid chromatography (HPLC), and the method agreements were, in general, good.     

The effect of a porcine-derived small intestinal submucosa product on wounds with exposed bone in dogs

OBJECTIVE: To determine the effect of a porcine-derived small intestinal submucosa product (PSIS) on healing time, epithelialization, angiogenesis, contraction, and inflammation of wounds with exposed bone on the distal aspect of the limbs of dogs. STUDY DESIGN: Prospective, controlled, experimental study. ANIMAL POPULATION: 10 young adult, purpose-bred, male Beagles. METHODS: Small wounds with exposed bone were created on the lateral aspect of metatarsal V and the medial aspect of metatarsal II on both hindlimbs. Three sheets of PSIS were sutured into the wounds of the treated limb, and the other limb served as a control. On day 10, punch biopsies of the medial metatarsal wounds were collected and were evaluated microscopically after routine hematoxylin and eosin and phosphotungstic acid hematoxylin (PTAH) staining. The lateral metatarsal wounds were evaluated by planimetry and laser Doppler perfusion imaging on days 7, 14, and 21. Time until complete wound healing was also recorded. The level of significance was set at P < or =.05 for all statistical analyses. RESULTS: Laser Doppler perfusion measurements were significantly higher in control wounds on day 7, but no differences were noted on days 14 and 21. No significant differences in planimetric values, histopathologic appearance, or time until complete wound healing were noted among treated and control groups. CONCLUSIONS: No objective differences in healing were noted between control wounds and wounds treated with PSIS. CLINICAL RELEVANCE: There appears to be no contraindication to the use of PSIS on clean wounds with exposed bone on the distal limbs of dogs. However, our objective data provides no evidence that this product affects epithelialization, contraction, or time to complete healing in wounds with exposed bone.     

Sedative and cardiopulmonary effects of medetomidine hydrochloride and xylazine hydrochloride and their reversal with atipamezole hydrochloride in calves

OBJECTIVE: To assess the sedative and cardiopulmonary effects of medetomidine and xylazine and their reversal with atipamezole in calves. ANIMALS: 25 calves. PROCEDURES: A 2-phase (7-day interval) study was performed. Sedative characteristics (phase I) and cardiopulmonary effects (phase II) of medetomidine hydrochloride and xylazine hydrochloride administration followed by atipamezole hydrochloride administration were evaluated. In both phases, calves were randomly allocated to receive 1 of 4 treatments IV: medetomidine (0.03 mg/kg) followed by atipamezole (0.1 mg/kg; n = 6), xylazine (0.3 mg/kg) followed by atipamezole (0.04 mg/kg; 7), medetomidine (0.03 mg/kg) followed by saline (0.9% NaCl; 6) solution (10 mL), and xylazine (0.3 mg/kg) followed by saline solution (10 mL; 6). Atipamezole or saline solution was administered 20 minutes after the first injection. Cardiopulmonary variables were recorded at intervals for 35 minutes after medetomidine or xylazine administration. RESULTS: At the doses evaluated, xylazine and medetomidine induced a similar degree of sedation in calves; however, the duration of medetomidine-associated sedation was longer. Compared with pretreatment values, heart rate, cardiac index, and PaO(2) decreased, whereas central venous pressure, PaCO(2), and pulmonary artery pressures increased with medetomidine or xylazine. Systemic arterial blood pressures and vascular resistance increased with medetomidine and decreased with xylazine. Atipamezole reversed the sedative and most of the cardiopulmonary effects of both drugs. CONCLUSIONS AND CLINICAL RELEVANCE: At these doses, xylazine and medetomidine induced similar degrees of sedation and cardiopulmonary depression in calves, although medetomidine administration resulted in increases in systemic arterial blood pressures. Atipamezole effectively reversed medetomidine- and xylazine-associated sedative and cardiopulmonary effects in calves.     

Functional markers delimiting a <Emphasis Type="Italic">Medicago</Emphasis> orthologue of pea symbiotic gene <Emphasis Type="Italic">NOD3</Emphasis>

Symbiotic gene mutated in the pea (Pisum sativum L.) line RisfixC is a determinant of the number of symbiotic root nodules. In parallel to a sharp increase in nodule number, its mutational inactivation brings about the insensitivity of nodulation to the ambient nitrate level (Nts trait). Using the established localization to the SYM2-NOD3 region of the pea linkage group I, functional PCR markers were developed for the orthologous region on the chromosome 5 of the model species Medicago truncatula. Owing to the conservation of the binding regions of the designed primers, pea orthologues were successfully amplified with 60% of the primer pairs tested. When applied to a mapping pea population from the cross of the line RisfixC x Afghanistan L1268 (sym2), the new markers allowed to localize the supernodulation mutation within 2.5 cM confidence interval in the pea genome. The placement of the functional markers on the M. truncatula chromosome 5 confined the orthologous gene location to eight overlapping BACs spanning approximately 710 kbp (positions 37,755,678–38,467,472). The narrowed list of the annotated Medicago genes in combination with the published data on their symbiotic and nitrate regulation can be used for the candidate gene identification, together with the requirements imposed by the known function in nodule number initiation and nitrate sensing. In addition, the new markers are applicable for tracking the RisfixC allele in breeding programmes aimed at the improvement of symbiotic performance.     

Genetic tracking of the brown bear in northern Pakistan and implications for conservation

Asian bears face major threats due to the impact of human activities as well as a critical lack of knowledge about their status, distribution and needs for survival. Once abundant in northern Pakistan, the Himalayan brown bear (Ursus arctos isabellinus) has been exterminated in most of its former distribution range. It presently occurs sparsely, in small populations, the Deosai National Park supporting the largest isolate. This decline might imply a reduction in genetic diversity, compromising the survival of the population. Using a combination of fecal DNA analysis and field data, our study aimed at assessing the size and genetic status of the Deosai population and give guidelines for its conservation and management. Using fecal genetic analysis, we estimated the population to be 40-50 bears, which compares well with the field census of 38 bears. The northern Pakistani brown bear population may have undergone an approximate 200-300-fold decrease during the last thousand years, probably due to glaciations and the influence of growing human population. However, in spite of the presence of a bottleneck genetic signature, the Deosai population has a moderate level of genetic diversity and is not at immediate risk of inbreeding depression. Gene flow might exist with adjacent populations. We recommend careful monitoring of this population in the future both with field observations and genetic analyses, including sampling of adjacent populations to assess incoming gene flow. The connectivity with adjacent populations in Pakistan and India will be of prime importance for the long-term survival of Deosai bears.     

Forest condition and management in Swedish forest commons

Forest commons are regarded as a means to support local development and sustainable forest conditions. To evaluate the development impact of Swedish forest commons, comparative surveys have been undertaken in three regions, and the differences in forest condition and management between categories of commons as well as their relation to other forest ownerships have been assessed. Regional differences between the by-laws, historical development and geographical conditions are apparent. It is concluded that two of three regions have an overly restrictive harvesting policy given the purpose of the forest commons and the official forest policy. The study results underline the importance of evaluation of the performance of forest management in relation to management objectives, to ownership alternatives and to the impact of local variations in preconditions.     

Cardiac arrhythmias and electrolyte disturbances in colic horses

Background

Despite increased focus on cardiac arrhythmias in horses, the nature and prevalence is still poorly described. Case reports suggest that arrhythmias occurring secondary to systemic disease are seen more commonly in the clinic than arrhythmias caused by cardiac disease. The aim of this study was to investigate the prevalence of arrhythmias in colic horses referred for hospital treatment. Associations between electrolyte disturbances and arrhythmias were also investigated.The study population consisted of eight control horses and 22 referred colic horses. A Holter electrocardiography (ECG) was recorded during the first 24 hours of admission. The ECG’s were analysed by a software program followed by manual visual inspection. Arrhythmias registered included second degree atrioventricular (AV) blocks, supraventricular premature complexes (SVPCs), and ventricular premature complexes (VPCs). Blood was collected at admission and again between 12 and 24 hours after ECG was applied, and analysed for concentrations of potassium, sodium, ionised calcium, chloride, glucose, and L-lactate.

Results

Heart rate was 37.4 ± 3.7 bpm in the control group, and 51.6 ± 11.8 bpm, in the colic group, which was significantly different (P < 0.0001). AV blocks and SVPCs were found in both groups, however only colic horses showed VPCs. No significant difference between the two groups was found for AV blocks, SVPCs, and VPCs (P = 0.08 - 0.76). The mean levels of potassium, sodium, ionized calcium, and chloride were significantly lower in the colic group compared to the control group at admission. Mean levels of glucose and L-lactate were significantly elevated in the colic group (P < 0.05).

Conclusions

This study describes prevalence of cardiac arrhythmias and electrolytes concentrations in colic horses compared to healthy controls. Although we only observed VPCs in the colic horses, no significant differences between colic horses and controls were found. Despite the colic horses having electrolyte changes at admission no correlation was found between the electrolyte disturbances and cardiac arrhythmias. Although no clear conclusions can be drawn from the present study, the results indicate that relatively mild colic per se is not pro-arrhythmogenic, whereas severe colic probably are more likely to result in ventricular arrhythmia.