Sewage sludge toxicity assessment using earthworm Eisenia fetida: can biochemical and histopathological analysis provide fast and accurate insight?

Sewage sludge (SS) is a complex organic by-product of wastewater treatment plants. Deposition of large amounts of SS can increase the risk of soil contamination. Therefore, there is an increasing need for fast and accurate assessment of SS toxic potential. Toxic effects of SS were tested on earthworm Eisenia fetida tissue, at the subcellular and biochemical level. Earthworms were exposed to depot sludge (DS) concentration ratio of 30 or 70 %, to undiluted and to 100 and 10 times diluted active sludge (AS). The exposure to DS lasted for 24/48 h (acute exposure), 96 h (semi-acute exposure) and 7/14/28 days (sub-chronic exposure) and 48 h for AS. Toxic effects were tested by the measurements of multixenobiotic resistance mechanism (MXR) activity and lipid peroxidation levels, as well as the observation of morphological alterations and behavioural changes. Biochemical markers confirmed the presence of MXR inhibitors in the tested AS and DS and highlighted the presence of SS-induced oxidative stress. The MXR inhibition and thiobarbituric acid reactive substance (TBARS) concentration in the whole earthworm's body were higher after the exposition to lower concentration of the DS. Furthermore, histopathological changes revealed damage to earthworm body wall tissue layers as well as to the epithelial and chloragogen cells in the typhlosole region. These changes were proportional to SS concentration in tested soils and to exposure duration. Obtained results may contribute to the understanding of SS-induced toxic effects on terrestrial invertebrates exposed through soil contact and to identify defence mechanisms of earthworms.

Allergenicity and immunogenicity of the major mugwort pollen allergen Art v 1 chemically modified by acetylation.

BACKGROUND: Treating allergies with modified allergens is an approach to make the treatment safer and more efficient. Art v 1 is the most prominent allergen of mugwort pollen and a significant cause of hayfever around Europe. The aim of this study was to reduce the allergenicity of Art v 1 by acetylation, and to investigate the capacity of the modified protein to generate blocking antibodies. METHODS: The reduction of allergenicity of Art v 1 following acetylation was monitored by immunoblot, ELISA inhibition using a pool of sera from mugwort pollen allergic patients, basophil activation assay and by skin prick testing of mugwort-allergic patients. Rabbits were immunized against Art v 1 and acetylated Art v 1 (acArt v 1) and the rabbit antisera were tested for their capacity to block human IgE binding in ELISA. Human T cell proliferation against Art v 1 and acArt v 1 was examined in peripheral blood mononuclear cells (PBMCs) of mugwort pollen allergic patients and cytokine release in PBMC cultures was monitored. RESULTS: Acetylation of Art v 1 gave a derivative of reduced allergenicity in the in vitro and ex vivo tests applied. The skin test reactivity to acArt v 1 was significantly reduced in 19 patients when compared with the reactivity to Art v 1. Rabbit antibodies to acArt v 1 and Art v 1 showed similar capacity to block human IgE binding to Art v 1 in inhibition ELISA. Both proteins were able to induce proliferation of PBMCs and CD3/CD4(+) cells of mugwort-allergic patients. Release of IL-5 was significantly reduced in cultures stimulated with acArt v 1. CONCLUSIONS: Art v 1 modified by acetylation had a significantly reduced allergenicity in vitro and in vivo, while its immunogenicity was retained. Modification of allergens by acetylation could be a new strategy for allergen-specific immunotherapy.

Classification accuracy of algorithms for blood chemistry data for three aquaculture-affected marine fish species.

The objective of this study was determination and discrimination of biochemical data among three aquaculture-affected marine fish species (sea bass, Dicentrarchus labrax; sea bream, Sparus aurata L., and mullet, Mugil spp.) based on machine-learning methods. The approach relying on machine-learning methods gives more usable classification solutions and provides better insight into the collected data. So far, these new methods have been applied to the problem of discrimination of blood chemistry data with respect to season and feed of a single species. This is the first time these classification algorithms have been used as a framework for rapid differentiation among three fish species. Among the machine-learning methods used, decision trees provided the clearest model, which correctly classified 210 samples or 85.71%, and incorrectly classified 35 samples or 14.29% and clearly identified three investigated species from their biochemical traits.

Nuclear abnormalities of marine fish erythrocytes.

The aim of this study was to monitor erythrocyte nuclear abnormalities (NA) including micronuclei (MN) in cultured and wild sea bass Dicentrarchus labrax and wild mullet Mugil spp. Seasonal sampling was performed at seven locations along the eastern coast of the Adriatic Sea. The frequency of NA and MN was positively correlated to temperature (NA: P < 0.05, r = 0.11; MN P < 0.05, r = 0.10), and there was also a positive correlation between NA and MN frequency (P < 0.001, r = 0.43). The lowest NA and MN values for both fish species were recorded in spring, while the highest were recorded in autumn. Significantly higher frequency of NA was seen in D. labrax compared to Mugil spp., while MN frequency was low in both species and not significantly different. There was no significant difference in NA and MN frequency between cultured and wild D. labrax sampled in the same month, and there was no difference between wild Mugil spp. sampled near or far from fish farms. In view of sampling sites, the highest values were detected in fishes from the Limski Channel, the lowest from the Janjina location.

Blood chemistry and histological properties of wild and cultured sea bass (Dicentrarchus labrax) in the North Adriatic Sea.

Histological and biochemical procedures were employed to study liver histomorphology and plasma levels of aspartate aminotransferases (AST), alanine aminotransferase (ALT), triglyceride (TRIG), cholesterol (CHOL), glucose (GLU) and total protein (TP) in cultured (CSB) and wild sea bass (WSB) (Dicentrarchus labrax) from the northern part of the Adriatic Sea. Histopathological changes in liver included varying degrees of infiltration and lipid degeneration of hepatocytes in examined cultured fish. No significant differences between median AST values of CSB (44 IU) and WSB (45 IU) were observed. Values of ALT were <5 IU in both groups. TRIG, CHOL, GLU and TP levels were higher in CSB (2.08 mmol/L, 3.67 mmol/L, 10.66 mmol/L and 49 g/L, respectively) than in WSB (0.67 mmol/L, 2.74 mmol/L, 3.68 mmol/L and 36 g/L, respectively). The study showed differences between plasma biochemical parameters and liver histomorphology of CSB and WSB. This can be explained as a consequence of different diets (artificial in contrast to natural foods), which influence energy metabolism.

Comparison of the API 20E and BBL Crystal E/NF identification systems for differentiating bacterial isolates from apparently healthy reared sea bass (Dicentrarchus labrax).

The ability of two commercial rapid identification systems, API 20E and BBL Crystal E/NF, to reliably identify bacterial isolates from the internal organs of reared sea bass were compared. The tests gave different results: API 20E identified bacteria as Pseudomonas spp. with 37% accuracy, while BBL Crystal E/NF identified them as Flavobacterium odoratum with 99% accuracy. Although F. odoratum is not a marine fish pathogen, conventional tests conducted with the same isolates were more indicative of them being Flavobacterium spp. than Pseudomonas spp., suggesting that BBL Crystal E/NF was more reliable in this identification. Both systems were found to be applicable for diagnostics of marine fish pathogens, but should be used with caution because of possible misinterpretation.

[Orbito-ocular dermoid cysts--incidence and particular cases]. / Chisturi dermoide oculo-orbitare incidenta si cazuri particulare.

This paper is concerned about a retrospective study on the casuistry of the Ophthalmologic Clinic of Craiova concerning those cases hospitalized and operated for different dermoidal cystic localization. A period of 20 years is under observation. The group of patients is analyzed taking into account the apparent debut age of the tumors, sex and different localization incidence. Both those tumor particular forms are exemplified by the clinic own casuistry and the association to other lesion types and profound localization in the orbit with particular clinic symptomatology. All the cases have been checked up and anatomopathologically confirmed.

[Less frequent etiology in uveitis]. / Uveitele cu etiologie mai putin frecventa.

This material intends to present three clinical observations regarding the uveitis etiology, that is represented by the association of some etiological factors less known: Toxoplasma gondii, Listeria monocytogenes, Leptospira icterohaemorrhagiae. The uveitis described here have been severe in the fast evolution and with trends of reappearence. This work demonstrates that the strict etiological antibacteria and antiparasite therapy is not enough and only corticotherapy is able to fight against the accentuated inflammatory phenomena that go along with type of uveitis.

Aeromonas hydrophila isolated from wild freshwater fish in Croatia.

Aeromonas hydrophila was recovered from fish living in lake Vrana on the Croatian island of Cres. The occurrence of the bacterium in the fish was assessed and related to gross signs of disease and findings at necropsy as a potential health hazard for fish. Isolated bacteria were subjected to morphological, physiological, biochemical and antibiotic susceptibility tests. A total of 26 A. hydrophila isolates were obtained. There was a clear seasonality, as no isolates were recovered in the summer months. Most of the isolates were sensitive to all the antimicrobials used in the study except novobiocin and penicillin G. Affected fish manifested haemorrhages over the skin, in the liver, kidney and swim bladder, spleen infarcts, fatty liver, ascitic fluid and swollen haemopoietic tissues. A. hydrophila does not appear to pose a major threat for the fish in the lake at present but under unfavourable and stressful conditions it could seriously compromise fish health.

Vibriosis in rainbow trout cultured in the Krka estuary, Croatia: occurrence and comments.

The frequency of vibriosis in cultured rainbow trout, maintained under different rearing conditions in the Krka estuary, was examined over a 6 yr period. Annual studies commenced regularly in October and ended in June of the following year. Every month during study periods, 37-75 trouts were randomly taken from each of 4 farm sites for routine examination. Twenty fish from these samples were subsequently employed for bacteriological analyses. Based on morphological and biochemical properties, the bacterial isolates were identified as Vibrio anguillarum, (biotype I). The findings demonstrate high occurrence of vibriosis in trout cultivated in the Krka estuary. Furthermore, there was a direct relationship between water quality parameters and the severity of vibriosis epizootics. Moreover, the causative agent has been isolated from free-living species, fish, notably eel and mullet, which are abundant to the Krka aquatorium. The findings from these long-term studies will be considered with reference to the developing salmon farming industry of Croatia.