Optimization of a culture technique for the isolation of Listeria monocytogenes from faecal samples

A culture technique employing cold enrichment at 4 degrees C followed by selective enrichment and plating at higher temperatures (30 degrees C) was used to isolate Listeria monocytogenes from faecal samples. The samples were held at 4 degrees C for 15 weeks and cultured weekly to assess the sensitivity of the culture after cold storage for different lengths of time. No media, Listeria selective enrichment broth (LSEB), nutrient broth (NB) and saline were used as cold storage medium. Cold storage increased the frequency of Listeria positive samples. The sensitivity of the culture for Listeria spp. and L. monocytogenes was 72 and 94%, and 56 and 61% after third and seventh week of cold storage, respectively. When the results of third and seventh week of cold storage were combined, the sensitivity was 100% for Listeria spp. and 94% for L. monocytogenes. LSEB and NB as storage medium increased Listeria positive samples after the first week of cold storage but did not maintain the increase thereafter while saline had an adverse effect on the growth of the bacteria. However, samples held in no media in a pilot study involving monthly sampling of a herd revealed better results. Detection limit of the culture media was also investigated. The lowest concentration detected by culture media was 3.17 organisms/ml. This was seven organisms/g for known Listeria positive sample. The faecal samples spiked with 10-fold dilutions of L. monocytogenes and held at 4 degrees C revealed that the sample spiked with 3.17 x 10-1 cfu/ml organisms resulted in growth after the second week of cold storage. The results suggest that the culture technique employing cold enrichment followed by selective enrichment and plating is more sensitive, the storage of faecal samples in no media when compared with the samples in storage medium, LSEB, NB and saline, during cold enrichment is a better application and culture of faeces, immediately after collection, at third and seventh week of cold enrichment produce more satisfactory results.     

Anatomical Study of the Gastrointestinal Tract in Free‐living Axis Deer (Axis axis)

The macroscopic anatomy of the stomach and intestines of adult axis deer (Axis axis), a cervid species considered intermediate/mixed feeder, was observed and recorded. Nine adult wild axis deers of both sexes were used and studied by simple dissection. The ruminal papillae were distributed unevenly in the overall area of the inner surface of rumen and primarily were more large and abundant within the atrium. The ruminal pillars had no papillae. There was an additional ruminal pillar located between the right longitudinal and right coronary ventral pillars connected to the caudal pillar. No dorsal coronary pillars were found, and the ventral coronary pillars are connected. The reticulum was the third compartment in size, and the maximum height of the reticular crests was 1.0 mm. The Cellulae reticuli were not divided and rarely contained secondary crests. There were no Papillae unguiculiformes. The omasum was the smallest gastric compartment. The abomasum had about twelve spiral plicae, and a small pyloric torus was present. The intraruminal papillation was similar to those species that are characterized by a higher proportion of grass in their natural diet. The finding of the small reticular crests is typical for browser ruminants and was coincident with data reported for other deer. The comparative ratio of the small intestine to the large intestine was 1.69, in terms of length measurements in axis deer and appears below of the ‘browser range’. We concluded that the gastrointestinal system of axis deer reflected similar morphological characteristics of the both types of ruminants: browser and grazer, and we consider it as an intermediate feeder.     

Dose-dependent immunohistochemical and ultrastructural changes after oral methylphenidate administration in rat heart tissue

Methylphenidate, more commonly known as Ritalin, is a piperidine derivative and is the drug most often used to treat attention deficit/hyperactivity disorder, one of the most common behavioural disorders of children and young adults. Our aims were to investigate dose-dependent immunohistochemical D2 expression and ultrastructural changes of the rat heart tissue, and to demonstrate possible toxicity of the long-term and high dose use of the methylphenidate. In this study, 27 female pre-pubertal Wistar albino rats, divided into three different dose groups (5, 10 and 20 mg/kg) and their control groups, were used. They were treated orally with methylphenidate dissolved in saline solution for 5 days/week during 3 months. At the end of the third month, after perfusion fixation, left ventricle of cardiac tissue was removed. Paraffin, semi-thin and thin sections were collected and immunohistochemical, terminal deoxynucleotidyl transferase-mediated Dig-dUTP nick end labelling assay and ultrastructural studies were performed. In conclusion, we believe that Ritalin is dose-related affecting dopaminergic system to increase heart rhythm and contraction. Thus, this drug may cause degenerative ultrastructural changes in mitochondrial path.     

Age-related immunohistochemical and ultrastructural changes in rat oculomotor nerve

During ageing process, multiple changes occur on nervous tissue composed of cells and extracellular matrix. Changes on nervous tissues are usually known as degenerative changes on axon structure and connective tissue covering the nerve such as a decrease in the number of fibre or general structural changes. For this purpose, we have studied age-dependent ultrastructural changes in the rat oculomotor nerve with electron microscopy and also demonstrated collagen structure of the neural sheaths with immunohistochemical techniques. This study was conducted in Gazi University Faculty of Medicine, Department of Anatomy with a total of nine Wistar albino rats. We observed strong collagen type I immunoreactivity in endoneurium and slight to moderate reactivity in fibroblast cytoplasm in 3-month- and 12-month-old groups and mild reactivity in 24-month-old group. Collagen type IV immunoreactivity was stronger in endoneurium and perineurium in the 3-month- and 12-month-old groups compared with collagen type I and fibroblast cytoplasm showed a very strong reactivity. On the other hand, in the 24-month-old group, there was slight reactivity in endoneurium and a strong reactivity in perineurium. NGF staining showed moderate to strong reactivity on Schwann cells of the 3-month-old group. The immunoreactivity decreased in the 12-month- and 24-month-old groups. In the 3-month-old rat group, Schwann cell cytoplasm, mitochondrial structure and neurofilaments were normal. In the 12-month-old group, there were no changes in organelle distribution, mitochondrial structure and neurofilaments, but there was an increase in the connective tissue. An inconsiderable number of degenerated myelinated nerves were observed. We detected an important decrease in the collagen type I immunoreactivity, which could suggest that the endoneurium, perineurium and epineurium are less resistant to the age-related collagen loss and that the peripheral nerve is protected by a weaker barrier in the old group. The collagen type IV immunoreactivity was significantly decreased with age. NGF synthesis decreases with age because of Schwann cell structural degeneration or for different reasons. Thus, this could explain the diminished capacity of regeneration and damage of the myelination of the peripheral nerve.     

Potential for using the World Reference Base for Soil Resources to identify less favoured areas

The European Commission (EC) has adopted a resolution seeking to test a proposed set of eight soil and climate criteria that objectively assesses and delimits agricultural areas suffering from natural handicaps. In areas designated as ‘Less Favoured Areas’ (LFA) agricultural production is hampered by natural handicaps, for example difficult climatic conditions, steep slopes in mountain areas or poor soil productivity. To delineate LFAs, a set of factsheets were provided by the EC Joint Research Center for mapping the proposed common soil, climate and terrain criteria. The present study was performed to determine the suitability of the World Reference Base for Soil Resources, 2006 (WRB) soil classification system in terms of estimating LFA thresholds. It was concluded that the WRB soil categories are directly applicable for delineating areas affected by the LFA criteria for soil constraints Organic Soil and Vertic Properties. The WRB categories correctly defined areas constrained by Salinity and Sodicity, but since they are stricter, than the LFA criteria, further analysis is needed to delineate the larger areas corresponding to LFA criteria. The climate and terrain criteria of LFA are only broadly indicated by WRB categories.     

Potential and limitations of isozymes for chromosomal localization of resistance genes against barley mild mosaic virus (BaMMV)

Summary To assess the possibilities offered by isozymes to locate resistance genes against barley mild mosaic virus (BaMMV), the isozyme patterns of 19 barley (Hordeum vulgare L.) genotypes carrying genes different from ym4 were determined. Of the 15 isozyme systems tested, only three were polymorphic, namely aconitate hydratase, esterase, phosphogluconate dehydrogenase, providing markers on four of the seven barley chromosomes. Studies of F₂ progenies derived from three crosses between resistant genotypes and susceptible varieties failed to reveal linkage between resistance genes and isozymes. Another goal of the experiment was to study the linkage relationships between ym4 and the esterase locus (Est1-Est2-Est4). Our estimates of the recombination rate between these two loci (3.41 and 8.32%) were in the range of those reported between these esterases and one of the resistance genes of the Chinese variety Mokusekko 3.     

Coralligenous assemblages along their geographical distribution: Testing of concepts and implications for management

• 1. The coralligenous habitat was studied at the large Mediterranean scale, by applying a standardized, non‐destructive photo‐sampling protocol, developed in the framework of the CIGESMED project.
• 2. The results provided evidence to support the following statements: (a) the assemblage pattern is not homogeneously distributed across the four Mediterranean ecoregions studied (biotic gradients hypothesis); and (b) the assemblage pattern does not change significantly when the information is aggregated to higher taxonomic levels (taxonomic sufficiency hypothesis).
• 3. Surrogate taxonomic categories higher than species, such as genus and family, can be used to reveal the multivariate pattern of the coralligenous assemblages.
• 4. Although preliminary at the pan‐Mediterranean scale, these outcomes set the scene for future comparisons as more data sets become available but also for comparisons between taxonomic and functional patterns.