Cabbage (Brassica oleracea var. capitata) as possible indicator of wartime metal and metalloid contamination in eastern Croatia (ICP-MS method)

Biomonitoring of the local population and environmental monitoring in eastern Croatia have revealed abnormalities in metal and metalloid distribution that could be related to war activities during the 1990s. The goal of this study was to determine whether there are differences in the concentrations of metals and metalloids by comparing locations of high and low-intensity combat activity; we also evaluated a possible connection between metal contamination in soil and in humans. We sampled 14 locations and measured the concentrations of 20 war related metals and metalloids (Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Ni, Pb, Sb, Si, Sr, U, V and Zn). The results of principal components analysis showed two main clusters: locations Dopsin and Dalj (both characterized by high-intensity combat activity), where the concentrations of most elements (except Hg) were increased. Although the concentrations of metals and metalloids in cabbage samples collected in eastern Croatia did not exceed the maximum allowed values, the results of our study confirmed existance of environmental ‘hotspot’ with higher concentrations of war metals and metalloids. Our findings indicate that there is a possible common source and mechanism of transferring metals from the environment to the population.     

Molecular tracing of the transmission routes of bois noir in Mediterranean vineyards of Montenegro and experimental evidence for the epidemiological role of Vitex agnus‐castus (Lamiaceae) and associated Hyalesthes obsoletus (Cixiidae)

Epidemiological aspects and transmission routes of bois noir (BN), a grapevine yellows disease induced by ‘Candidatus Phytoplasma solani’, have been exhaustively studied in the affected vineyards of continental Europe but not in the Mediterranean coastal zone. Because ‘Ca. Phytoplasma solani’ and its principal vector Hyalesthes obsoletus presumably originate from the Mediterranean, gaining knowledge of the epidemiological peculiarities of the disease in this area is essential for understanding its global spread and diversification, as well as for designing local management strategies. In this study, molecular epidemiology was applied to trace transmission pathways of ‘Ca. Phytoplasma solani’ in the Mediterranean vineyards of Montenegro, using multilocus sequence typing of tuf, vmp1 and stamp genes of the isolates associated with various hosts. Thus, ‘Ca. Phytoplasma solani’ was tracked from a tentative reservoir plant (inoculum source) through an associated vector population to the infected grapevine. Three pathways of transmission were documented, originating from Urtica dioica, Convolvulus arvensis and Vitex agnus‐castus; however, only the route originating from U. dioica was direct, whereas the latter two were overlapping and could be intermixed. Vitex agnus‐castus is a natural source of ‘Ca. Phytoplasma solani’, representing an important link in disease epidemiology in the Mediterranean and a possible origin of several genotypes occurring in central Europe. Experimental confirmation of the role of Vitex‐associated H. obsoletus in BN transmission in Montenegrin vineyards indicates its tentative role as a vector in the wide area of the Mediterranean, where some of the major wine‐producing regions are located.     

Predicting the solubility of Cd,Cu, Pb and Zn in uncontaminated Croatian soils under different land uses by applying established regression models

Soil pH, soil organic matter (SOM), dissolved organic carbon (DOC) and total trace metal concentration (M_(tot)) control the solubility of metals in the soil. Several regression models have included these soil chemical variables for the prediction of metal solubility and free metal ion (FMI) concentrations in contaminated soils. We hypothesize that models developed on contaminated soils (after optimization of the coefficients) can be used on samples from uncontaminated sites. Soil samples were collected from unpolluted agricultural and forest soils located in Eastern Croatia and extracted with water to determine the concentrations of Cd, Cu, Pb and Zn. We used these data to test the applicability of three regression models on existing conditions under different land uses. The same predictors issued in the three models and the same regression coefficients were utilized in the present study. The results showed a good correlation between the observed and predicted values of metal solubility. However, the models overestimate the total solution concentration (M_(sol)) and the concentrations of free metal ions (FMI) in solution, and therefore the same regression coefficients were optimized to fit our own observations. This was found to be very successful. The results showed that pH and DOC played a very important role in controlling metal solubility, while SOM and CEC were somewhat less significant. The impact of total soil concentration of metals (M_(tot)) was rather minor. However, we feel that to carry out good predictions of M_(sol) and FMI, the M_(tot) is needed in such regression models.     

On determination of chitin content in mushrooms

Nitrogen compounds constitute the greatest proportion of the dry weight of mushrooms. These nitrogen compounds are partly protein and partly non-protein. A modified Heneberg-Stohman's method was used to separate non-protein from protein nitrogen, as well as to determine raw fiber content in mushrooms, since the methods used previously for this purpose are time-consuming and complicated. Elemental analysis of crude fiber was performed to confirm the uniformity of isolated products. The content of total crude fiber (fungin) and the proportion of chitin and cellulose in it were determined. The chitin content of 15 species of field grown edible mushrooms was in the range of 1.87–6.93% of total dry mass and 34.51–67.72% of isolated fungin.     

Microbiological identification and analysis of swine tonsils collected from carcasses at slaughter

The primary objective of this 7-month study was to determine the prevalence of porcine pathogens of the tonsil of the soft palate of swine at slaughter. Additional objectives were to determine if sampling the carcasses of normal or abnormal hogs provided different microbiological profiles and if the slaughter plant provides a feasible sampling frame and environment for detecting and monitoring important pathogens in tonsils that have health implications for both swine and humans. A total of 395 samples were collected from 264 farms. Of these, 180 tonsils were collected from normal carcasses and 215 tonsils were collected from carcasses that were diverted to the hold rail. Laboratory testing included bacteriological culture and identification as well as real time-polymerase chain reaction (PCR) testing for porcine reproductive and respiratory syndrome virus (PPRSV) and immunohistochemistry (IHC) for porcine circovirus-2 (PCV-2). The most commonly isolated bacteria included: Streptococcus suis (53.7%), Arcanobacterium pyogenes (29.9%), Pasteurella multocida (27.3%), and Streptococcus porcinus (19.5%). Virus screening revealed evidence of PRRSV and PCV-2 in 22.0% and 11.9% of the samples, respectively. Salmonella Typhimurium and Yersinia enterocolitica were isolated in 0.5% and 1.8% of the samples, respectively. Tonsils collected from the hold rail were more likely to be positive for Staphylococcus hyicus [odds ratio (OR) = 7.51, confidence interval (CI) = 2.89 to 19.54], Streptococcus porcinus (OR = 9.93, CI = 4.27 to 23.10), and Streptococcus suis (OR = 2.16, CI = 1.45 to 3.24). Tonsils collected from abnormal carcasses were less likely to be positive for Staphylococcus aureus (OR = 0.05, CI = 0.005 to 0.482).     

The presence of methicillin-resistant Staphylococcus aureus on large pig breeding farms in Croatia

Methicillin-resistant Staphylococcus aureus (MRSA) have emerged worldwide and have become resistant to a variety of antibiotics. MRSA colonisation in pigs was first reported from the Netherlands in 2005, where pigs were implicated as a source of human MRSA infections (Voss et al., 2005). This paper presents the first report on the presence of MRSA on large pig breeding farms in Croatia, together with the determination of the mecA gene, the results of spa typing and susceptibility to commonly used antimicrobials. Dust samples (7-11 per farm) were collected from eight large pig farms in Croatia. Of the total 68 swabs, the mecA gene was detected in 24 isolates growing on the MRSA agar. All isolates were resistant to oxacillin, tetracycline and streptomycin, and susceptible only to vancomycin, while 92% of the strains were susceptible to ciprofloxacin. Genotyping of the MRSA strains was performed by spa typing, and revealed t011 (n = 17), t034 (n = 5) and t1451 (n = 2). The results presented here predict that MRSA is present on a large number of pig farms in Croatia.     

Trace elements in soils and food chains of the Balkan region

Abstract

The present review summarizes the available data on the concentration of trace elements (TEs) in soils and their availability to plants with a view to reflect the quality and safety of food and fodder crops. Most soils in western Balkan countries are not contaminated. However, soils around industrial centers and historical mining sites do create concern for food and feed safety. Also high concentrations of TEs are related to their geochemical original. For example, ultrabasic rocks and serpentinites in western Serbia and western Bosnia are very rich in chromium, nickel, magnesium, iron (Fe), and cobalt, and cover an area of 5200 km². High TE concentrations caused by anthropogenic activities are also reported. In vineyard soils in Croatia, concentrations of cadmium, copper (Cu), and zinc (Zn) were much higher than their background concentrations. On the other hand, TE deficiency in plants is also prevalent in the regions. For example, Zn and Fe deficiencies in eastern parts of Croatia and northern parts of Serbia, Cu deficiency in pasture and sheep's blood at Nisici Plateau of Bosnia and Herzegovina, and selenium (Se) deficiency almost in the whole region have been observed. Therefore, information on TEs' behavior and soil factors affecting their mobility and availability is highly needed in order to separate the areas of contamination and then propose agrotechnical measures to protect the entry of TEs into the food chain. Research is also required to assess the influence of agronomic management on TE supply to plants and for achieving a better utilization of essential TEs. Concentrations of Se in wheat in Serbia are so low that if people were fed exclusively with wheat, their daily requirements for Se would not be met. There is also a need for full implementation of new food safety regulations in the Balkan countries in accordance with the legislations of the European Union.     

Growth Responses,Metal Accumulation and Phytoremoval Capability in Amaranthus Plants Exposed to Nickel Under Hydroponics

Surfactant-enhanced solubilization and subsequent biodegradation of phenanthrene and pyrene from aqueous solutions by Arthrobacter strain Sphe3 was investigated. The results show that growth of Arthrobacter strain Sphe3 was increased upon increase in concentration of Tween 20 and Tween 80. Inhibition of bacterial growth was observed with increasing Triton X-100 concentrations, whereas sodium dodecyl sulfate (SDS) totally inhibited this bacterial growth. Phenanthrene and pyrene solubilization was enhanced in the presence of surfactants and found to be linearly proportional to their concentrations, above the critical micelle concentration (CMC). In addition, Tween 20 and Tween 80 enhanced the biodegradation of phenanthrene and pyrene. The high correlation coefficient (R ²) values obtained at all the concentrations studied, suggest that biodegradation kinetics of both phenanthrene and pyrene in the presence of Tween 20 and Tween 80 follow first-order kinetic equation model. Experimental results suggest that Tween 20 and Tween 80 may have great potential for applications in bioremediation of these polycyclic aromatic hydrocarbon (PAH) compounds using Arthrobacter strain Sphe3.