Sentinel lymph node mapping by near‐infrared fluorescence imaging and contrast‐enhanced ultrasound in healthy dogs

Sentinel lymph node (SLN) mapping is a valuable and crucial diagnostic procedure in staging malignancies. We compared two non‐invasive techniques, near‐infrared (NIR) fluorescence imaging and contrast‐enhanced ultrasound (CEUS), to identify the SLNs in three superficial anatomical regions in an animal model. Six healthy laboratory dogs were included in a proof‐of‐concept trial. A NIR fluorescent dye (Indocyanine Green) and microbubbles (Sonovue) were consecutively injected subdermally in the Inguinal, axillary and popliteal region to map the SLNs. Transcutaneous NIR fluorescence imaging identified SLNs in 17 out of a total of 18 occasions. CEUS identified SLNs in all regions (18/18). Whereas NIR fluorescence imaging performed better in the visualization of the afferent lymphatic tract, CEUS demonstrated different filling patterns of the SLNs, a feature potentially critical for the concept of SLN mapping in cancer patients. Both NIR fluorescence imaging and CEUS are safe, non‐invasive, practical and accurate methods to perform real‐time transcutaneous SLN mapping with potential in a clinical setting.     

Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

Assessment of infectious organisms associated with chronic rhinosinusitis in cats

OBJECTIVE: To determine detection rates for feline herpesvirus type 1 (FHV-1), Mycoplasma spp, fungi, and bacteria in flush samples and biopsy specimens from the nasal cavities of cats with and without chronic rhinosinusitis (CRS). DESIGN: Prospective study. ANIMALS: 10 CRS-affected cats and 7 cats without signs of respiratory tract disease. PROCEDURES: Nasal flush samples and biopsy specimens were collected from all cats for bacterial (aerobic and anaerobic), fungal, and mycoplasmal cultures; additional biopsy specimens were collected for virus isolation and polymerase chain reaction (PCR) assay (to detect FHV-1 DNA). RESULTS: Aerobic bacteria were detected in flush samples from 5 of 7 control cats; culture of flush samples from CRS-affected cats yielded aerobic bacteria (9/10 cats), anaerobic bacteria (3/10), and Mycoplasma spp (2/10). No fungal organisms were isolated from any cat. Potential pathogens were isolated significantly more often from CRS-affected cats than from control cats. Bacterial culture of biopsy specimens yielded aerobic bacteria (2/7 control cats and 4/10 CRS-affected cats) and anaerobic bacteria (2/10 CRS-affected cats). Although FHV-1 was not detected in nasal biopsy specimens from control or CRS-affected cats, FHV-1 DNA was detected via PCR assay in specimens from 4 of 7 control cats and 3 of 10 CRS-affected cats. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with findings in control cats, anaerobic bacteria, Mycoplasma spp, and a variety of potentially pathogenic organisms were detected more commonly in samples from cats with CRS. In both groups, FHV-1 was detected via PCR assay as a nonviable organism or in noncultivable amounts.     

RAPD analysis of interspecific relationships in presumably apomictic Cotoneaster species

A few of the approximately 300 Cotoneaster species described are diploid but the majority appear to be polyploid. The occurrence of apomixis inpolyploid Cotoneaster species has been reported but never proven with genetic markers. We have used 76 polymorphic RAPD markers to investigate the breeding system and phenetic grouping of some critical taxa, including a total of 19 plant accessions representing 13 mostly European species in the series Cotoneaster. Three to four individual plants, raised from seed from the same original plant, were analyzed for each of three accessions to investigate the possible occurrence of apomictic seed set. Absolutely congruent RAPD profiles were encountered among seedlings from one accession, whereas we found one or two marker differences among seedlings from the other two accessions. Genetic similarities among the different accessions were analyzed with a UPGMA-derived dendrogram. The most deviant taxon was the Chinese C. albokermesinus. A group withC. soczavianus and C. tomentosus was rather isolated from the remainder, as was also C. kullensis. Among the remaining taxa, two well supported clusters were found: (1) C. antoninae and C. uralensis, and (2) C. integerrimus and C. raboutensis, whereas the other five species (C. canescens, C. niger, C. scandinavicus, C. juranus, C. cambricus) formed a poorly supported cluster with no clear substructuring. A principal coordinate analysis yielded results that were in good correspondence with the dendrogram. Again C. albokermesinus appears to be totally isolated from the other species. In addition, two well-defined and rather isolated groups were found: (1) C. tomentosus and C. soczavianus, and (2) C. antoninae and C. uralensis, with the remainder comprising a loosely defined group. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of glucose in removal of microcystin-LR by viable commercial probiotic strains and strains isolated from dadih fermented milk

The removal of the cyanobacterial peptide toxin microcystin-LR at 4 and 37 degrees C by six commercial probiotic strains and Lactobacillus plantarum strains IS-10506 and IS-20506 isolated from dadih, Indonesian traditional fermented milk, was assessed in this study. The aim was to evaluate the main factors influencing the viability and metabolic activity of the probiotic strains, as well as their capacity to remove microcystin-LR. Both L. plantarum strains isolated from dadih, as well as Bifidobacterium lactis strains Bb12 and 420, were shown to be more resistant, and >85% remained viable in phosphate-buffered saline (PBS) solution for 48 h of incubation at either temperature, while the viability of the other four commercial bacteria decreased markedly over time. The effect of glucose on viability and removal of toxin was shown to be a strain-specific and strain-dependent property, but in general, the efficiency of microcystin-LR removal increased when glucose was added to the solution. A maximum removal of 95% was observed for L. plantarum strain IS-20506 (37 degrees C, 10 (11) colony-forming units mL(-1)) with 1-2% glucose supplementation and 75% in PBS alone.     

Bioindication and modelling of atmospheric deposition in forests enable exposure and effect monitoring at high spatial density across scales

Key message

Moss surveys provide spatially dense data on environmental concentrations of heavy metals and nitrogen which, together with other biomonitoring and modelling data, can be used for indicating deposition to terrestrial ecosystems and related effects across time and areas of different spatial extension.

Context

For enhancing the spatial resolution of measuring and mapping atmospheric deposition by technical devices and by modelling, moss is used complementarily as bio-monitor.

Aims

This paper investigated whether nitrogen and heavy metal concentrations derived by biomonitoring of atmospheric deposition are statistically meaningful in terms of compliance with minimum sample size across several spatial levels (objective 1), whether this is also true in terms of geostatistical criteria such as spatial auto-correlation and, by this, estimated values for unsampled locations (objective 2) and whether moss indicates atmospheric deposition in a similar way as modelled deposition, tree foliage and natural surface soil at the European and country level, and whether they indicate site-specific variance due to canopy drip (objective 3).

Methods

Data from modelling and biomonitoring atmospheric deposition were statistically analysed by means of minimum sample size calculation, by geostatistics as well as by bivariate correlation analyses and by multivariate correlation analyses using the Classification and Regression Tree approach and the Random Forests method.

Results

It was found that the compliance of measurements with the minimum sample size varies by spatial scale and element measured. For unsampled locations, estimation could be derived. Statistically significant correlations between concentrations of heavy metals and nitrogen in moss and modelled atmospheric deposition, and concentrations in leaves, needles and soil were found. Significant influence of canopy drip on nitrogen concentration in moss was proven.

Conclusion

Moss surveys should complement modelled atmospheric deposition data as well as other biomonitoring approaches and offer a great potential for various terrestrial monitoring programmes dealing with exposure and effects.

     

Lignin degradation by Agaricus bisporus accounts for a 30% increase in bioavailable holocellulose during cultivation on compost

The common mushroom Agaricus bisporus is a non-white rot saphrophytic fungus that can degrade lignin to free and utilize holocellulose embedded in fermented straw as present in compost. A new method is described to estimate the actual amount of bioavailable holocellulose in 3.8 kg compost cultures spawned with A. bisporus Horst U1 prior to and during a cultivation with two cycles of mushroom harvesting. The method shows that the initial amount of bioavailable holocellulose per culture, accounting for 130 +/- 22 g, is lower than the total holocellulose consumption by A. bisporus accounting for 182 +/- 15 g. This difference is explained by a 30% increase in bioavailable holocellulose. The increase is caused by the degradation of 95 +/- 3 g of holocellulose-shielding lignin. The results are discussed within the scope of the A. bisporus mushroom yield and lignin degradation by white rot fungi during growth on lignocellulose-containing materials.