Layout proposal for a microtitre plate to be used in routine antimicrobial susceptibility testing of bacteria from infections of dogs and cats

The determination of minimum inhibitory concentrations by broth microdilution is recommended as method of choice for susceptibility testing of veterinary bacterial pathogens. Accordingly, broth microdilution is used in veterinary routine diagnostic laboratories at a progressive rate. To reduce the costs of susceptibility testing, it is reasonable to develop widely accepted uniform microtitre plate layouts that are produced in large quantities. Such microtitre plate layouts have already been developed and published for the susceptibility testing of pathogens from food-producing animals. However, a microtitre plate layout, especially designed for the testing of bacteria from dogs and cats, should be available, too. The choice of the antimicrobial agents or combinations of antimicrobial agents to be included in a suitable layout should be based on the following criteria: (1) the approval and availability of an antimicrobial agent or combination of agents, (2) known cross-resistances, and (3) availability of approved clinical breakpoints. The latter point is of particular importance for the choice of the numbers of concentrations per antimicrobial agent tested and the range of test concentrations. Taking into account these aspects, a science-based layout proposal for microtitre plates, which are suitable for routine testing of bacteria from dogs and cats, is presented and discussed.     

Selection of Culture Conditions and Cell Morphology for Biocompatible Extraction of β-Carotene from Dunaliella salina

Biocompatible extraction emerges recently as a means to reduce costs of biotechnology processing of microalgae. In this frame, this study aimed at determining how specific culture conditions and the associated cell morphology impact the biocompatibility and the extraction yield of β-carotene from the green microalga Dunaliella salina using n-decane. The results highlight the relationship between the cell disruption yield and cell volume, the circularity and the relative abundance of naturally permeabilized cells. The disruption rate increased with both the cell volume and circularity. This was particularly obvious for volume and circularity exceeding 1500 µm³ and 0.7, respectively. The extraction of β-carotene was the most biocompatible with small (600 µm³) and circular cells (0.7) stressed in photobioreactor (30% of carotenoids recovery with 15% cell disruption). The naturally permeabilized cells were disrupted first; the remaining cells seems to follow a gradual permeabilization process: reversibility (up to 20 s) then irreversibility and cell disruption. This opens new carotenoid production schemes based on growing robust β-carotene enriched cells to ensure biocompatible extraction.     

Quantitative structure–activity studies of insect growth regulators: XVI. Substituent effects of dibenzoylhydrazines on the insecticidal activity to Colorado potato beetle Leptinotarsa decemlineata

Insecticidal activity against the Colorado potato beetle, Leptinotarsa decemlineata, was measured for a series of substituted N-tert-butyl-dibenzoylhydrazines, in which one of the benzoyl moieties closer to the tert-butyl group was fixed as being 2-chloro-substitued and the other variously substituted singly or doubly. The effects of substituents on the activity were quantitatively analysed using the classical quantitative structure–activity relationship (QSAR) procedure. The activity against the Colorado potato beetle increases with the molecular hydrophobicity. In addition, various types of steric effect are at work, depending upon the positions. Hydrogen-bonding acceptor substituents at the para position enhance the activity. There seem to be threshold (or optimum) values, albeit position-dependent, in the molecular hydrophobicity, above which the activity starts to decrease. This biphasic contribution of the molecular hydrophobicity to activity against coleopterous larvae is the most conspicuous difference in substituent effects from those found for similar compounds against lepidopterous pest insects, and may be the basis of the variations in the activity spectrum for certain compounds in this series. The introduction of bulkier substituents into the meta- and para-positions of the benzene ring, apart from the tert-butyl group, is unfavorable to activity. LD₅₀ values against Colorado potato beetle larvae of methoxyfenozide (RH-2485) and tebufenozide (RH-5992) were in the order of 10⁻⁷ mol per insect, whereas those of RH-5849, and halofenozide (RH-0345) were very low, 10⁻⁹–10⁻¹⁰ mol per insect being selective to the coleopterous larvae. © 1999 Society of Chemical Industry     

Accelerated Degradation and Mineralization of Atrazine in Surface and Subsurface Soil Materials

In surface soils, atrazine is considered to be a moderately persistent herbicide, with half-lives ranging generally from one to two months. In subsoils, however, its degradation is generally slower. This paper reports the degradation of atrazine in soil and subsoil samples taken from six Belgian maize fields. Rapid degradation can take place in some samples taken from surface and in some from subsurface soils. Subsoil samples were found to degrade atrazine either very strongly or not at all. Experiments with [ring-U-¹⁴C] atrazine showed that the micro-organisms responsible for the rapid degradation cleave the triazine ring and extensively mineralize the molecule. © 1997 SCI.     

Live prey first feeding regimes for short‐snouted seahorse Hippocampus hippocampus (Linnaeus, 1758) juveniles

As with many species of seahorses, Hippocampus hippocampus wild populations are being subjected to uncontrolled exploitation in their natural environment. Thus, aquaculture could contribute to satisfy the commercial demand for animals while promoting the recovery of wild stocks. The present study was conducted to compare the effect of the substituting Artemia nauplii with rotifers for first feeding seahorse juveniles. Survival, growth and biochemical composition of prey organisms and fish were studied during the feeding trial. In addition, to help the biometric study, an anaesthetic test was also carried out using clove oil. The results showed excellent survival (average 60%) in juveniles exclusively fed with Artemia, with better values than those reported previously obtained by other authors for this species. By comparison, high mortality and poor growth were observed during first feeding with seahorses fed on rotifers. This could have been related to the lower energy intake and poorer nutritional value of the rotifers. Furthermore, clove oil concentrations of 25 ppm were found to work well as an anaesthetic for seahorse juveniles. Overall, first feeding Artemia alone was found to be an efficient and simplified method for feeding young H. hippocampus fry, building the principles for their culture for ornamental or re‐stocking purposes.     

Conservation agriculture as a sustainable option for the central Mexican highlands

Tropical highlands of the world are densely populated and intensively cropped. Agricultural sustainability problems resulting from soil erosion and fertility decline have arisen all over this agro-ecological zone. Based on selected soil quality indicators, i.e. time-to-pond, aggregate distribution and stability (expressed as the mean weight diameter (MWD) for dry and wet sieving, respectively) and soil moisture, from a representative long-term sustainability trial initiated in 1991 in Central Mexico (2240 masl; 19.31°N, 98.50°W; Cumulic Phaeozem), some insights into the feasibility of conservation agriculture (CA) as part of a sustainable production system in the tropical highlands are given. Zero tillage plots with crop residue removal showed low aggregate distribution (average MWD = 1.34 mm) and stability (average MWD = 0.99 mm) resulting in top layer slaking, increased erosion and low time-to-pond values. Retaining the residue in the field with zero tillage avoided the above-mentioned negative evolution for both aggregate distribution as stability (average MWD = 2.77 and 1.51 mm, respectively) and even improved the physical conditions of the soil as compared to conventional practice. Throughout the growing season the lowest soil moisture content was found in zero tillage without residue (average over the entire growing season = 20.5% volumetric moisture content), the highest in zero tillage with residue retention (average = 29.7%) while conventional tillage had intermediate soil moisture values (average = 27.4%). Zero tillage without residue retention had most days of soil moisture values under permanent wilting point, while zero tillage with residue retention had the least. Taking into account these results, zero tillage with residue retention can clearly be a part of an integrated watershed management scheme towards sustainable agriculture in the tropical highlands. It is clear that to develop new management practices to improve water use, reduce erosion and enhance human labor/animal power focus must be on the use of conservation agriculture both for rainfed as well as irrigated production systems and be fine tuned for each system.     

USE OF COMPUTED TOMOGRAPHY AND COMPUTED TOMOGRAPHIC MYELOGRAPHY FOR ASSESSMENT OF SPINAL TUMORAL CALCINOSIS IN A DOG

Spinal tumoral calcinosis is reported in a Berner sennenhund puppy. The condition was manifested clinically as a non-ambulatory tetraparesis associated with neck pain. On survey radiographs there was a focal calcified mass at the atlantoaxial articulation. Computed tomography and computed tomographic myelography gave additional information on the extent of the mass and on the degree of spinal cord compression. The mass was removed surgically and the dog made a complete recovery.     

Successful engraftment of cultured autologous mesenchymal stem cells in a surgically repaired soft palate defect in an adult horse

The objective of this study was to graft autologous mesenchymal stem cells (MSCs) at the site of surgical repair of a soft palate defect in an adult horse in an attempt to improve wound healing and to investigate whether the transplanted MSCs would integrate into the soft palate structure and participate in regeneration. Bone marrow was collected from an adult horse with a full-thickness soft palate defect. The MSCs were isolated, cultured in monolayers, and labeled with 5-bromo-2-desoxymidine (BrdU) and chloromethylbenzamido-DiI-derived (cm-DiI) before transplantation. The soft palate defect was repaired by mandibular symphysiotomy, and the labeled MSCs were injected into the repaired soft palate. Postmortem examination revealed that 90% of the soft palate defect had been sutured. Staining by BrdU and cm-DiI was intense in the soft palate tissue. Labeled MSCs were detected in tissue slices from the injection sites. The cells were organized in a manner similar to that in native soft palate tissue, indicating successful engraftment.     

Comparative acaricide susceptibility and detoxifying enzyme activities in field-collected resistant and susceptible strains of Tetranychus urticae

A field-collected strain (MR-VL) of the two-spotted spider mite, Tetranychus urticae Koch, exhibited strong resistance to bifenthrin, dicofol and fenbutatin oxide in comparison with a susceptible laboratory strain (LS-VL). The MR-VL strain was screened for cross-resistance with several currently used acaricides. Cross-resistance was detected with clofentezine (RR = 2631), dimethoate (RR = 250), chlorfenapyr (RR = 154), bromopropylate (RR = 25), amitraz (RR = 17), flucycloxuron (RR = 15) and azocyclotin (RR = 7). Abamectin, acequinocyl, bifenazate, tebufenpyrad and spirodiclofen did not show any signs of cross-resistance. Enhanced detoxification by increased activity of mono-oxygenases (MO) and esterases is at least partially responsible for the observed resistance and cross-resistance. MO assays with 7-ethoxycoumarin (7-EC) were optimised and 7-ethoxy-4-trifluoromethylcoumarin (7-EFC), a new MO-substrate, was evaluated for the first time in T urticae and proved to be a good alternative to 7-EC. Approximately 3- and 4-fold higher MO activity was detected with 7-EFC and 7-EC respectively in the MR-VL strain. Kinetic parameters of general esterase assays with 4-nitrophenyl acetate and 1-naphthyl acetate as substrate indicated that more esterases were present in the MR-VL strain. A first attempt was made to classify the esterases present in T urticae. Acetyl-, aryl- and carboxyl-esterases were detected with the use of inhibitors after separation by native PAGE. Glutathione-S-transferases did not seem to play any role in the observed resistance and no differences were detected when the general oxidative capacities of the two strains were compared.