Cytokinin signalling regulates organ identity via the AHK4 receptor in Arabidopsis.

Mutual interactions of the phytohormones, cytokinins and auxin determine root or shoot identity during postembryonic de novo organogenesis in plants. However, our understanding of the role of hormonal metabolism and perception during early stages of cell fate reprogramming is still elusive. Here we show that auxin activates root formation, whereas cytokinins mediate early loss of the root identity, primordia disorganisation and initiation of shoot development. Exogenous and endogenous cytokinins influence the initiation of newly formed organs, as well as the pace of organ development. The process of de novo shoot apical meristem establishment is accompanied by accumulation of endogenous cytokinins, differential regulation of genes for individual cytokinin receptors, strong activation of AHK4-mediated signalling and induction of the shoot-specific homeodomain regulator WUSCHEL. The last is associated with upregulation of isopentenyladenine-type cytokinins, revealing higher shoot-forming potential when compared with trans-zeatin. Moreover, AHK4-controlled cytokinin signalling negatively regulates the root stem cell organiser WUSCHEL RELATED HOMEOBOX 5 in the root quiescent centre. We propose an important role for endogenous cytokinin biosynthesis and AHK4-mediated cytokinin signalling in the control of de novo-induced organ identity.

Spatial and Temporal Trends in Contamination of the Czech Part of the Elbe River by Mercury Between 1991 and 2016.

We summarized available data concerning contamination of the Czech part of the Elbe River by mercury and assessed the potential risks to humans connected with the consumption of contaminated fish. Total mercury concentrations were evaluated in dorsal muscle of chub (Squalius cephalus) and bream (Abramis brama) that were collected at 11 sampling sites in the period from 1991 to 2016. Significant differences among sites were found for both species. The highest contamination was observed near the non-ferrous Kovohute metal works at Celakovice and also downstream from the Spolana chemical plant at Neratovice. A significant decline in contamination with time (concentrations decreasing from 1.65 to 0.22 mg kg-1; p = 0.004) was revealed in chub downstream from the Spolana plant. Calculated hazard indices (HIs) were below 1, therefore, adverse health effects on the general Czech population connected with the consumption of fish from the Elbe River were not expected.

Effects of selenium feed supplements on functional properties of eggs.

The aim of the present study was to assess the effects of selenium feed supplements on the functional properties of eggs. The hens in experimental groups were fed diets supplemented with 0.2 mg/kg selenium from sodium selenite (Group 1), selenium-enriched yeast (Group 2), synthetic L-selenomethionine (Group 3), or hydroxy analog of selenomethionine (Group 4). The Control Group (Group C) was fed with basal feed without supplementation. The highest values of albumen gel firmness were shown in Group C eggs; differences with all experimental groups were significant (p < 0.001 to p = 0.009). It was ascertained that albumen gel firmness correlated with albumen pH (rs = 0.490; p < 0.001), which was highest in eggs from non-supplemented hens. Group 1 eggs and Group C eggs showed lower albumen foaming capacity (p < 0.001) compared to eggs from other groups. Both albumen foaming capacity and albumen foam stability were higher in Group 2 eggs than in Group C eggs (p < 0.001). The highest yolk foaming capacity was found in Group 2 eggs (p < 0.001). Sponge cakes baked with Group C eggs had a smaller volume than those baked with eggs from Group 2 (p = 0.005), Group 3 (p = 0.004) and Group 4 (p = 0.024). The results of the study confirmed that selenium added to the laying hen feed significantly affected the monitored functional properties of both albumen and yolk. The most distinctive effect of selenium was shown in eggs from the group supplemented with selenium-enriched yeast, for which the results of albumen foam capacity and stability and yolk foaming capacity were the best.

Antibody-mediated modulation of cytokinins in tobacco: organ-specific changes in cytokinin homeostasis.

Cytokinins comprise a group of phytohormones with an organ-specific mode of action. Although the mechanisms controlling the complex networks of cytokinin metabolism are partially known, the role of individual cytokinin types in the maintenance of cytokinin homeostasis remains unclear. Utilizing the overproduction of single-chain Fv antibodies selected for their ability to bind trans-zeatin riboside and targeted to the endoplasmic reticulum, we post-synthetically modulated cytokinin ribosides, the proposed transport forms of cytokinins. We observed asymmetric activity of cytokinin biosynthetic genes and cytokinin distribution in wild-type tobacco seedlings with higher cytokinin abundance in the root than in the shoot. Antibody-mediated modulation of cytokinin ribosides further enhanced the relative cytokinin abundance in the roots and induced cytokinin-related phenotypes in an organ-specific manner. The activity of cytokinin oxidase/dehydrogenase in the roots was strongly up-regulated in response to antibody-mediated formation of the cytokinin pool in the endoplasmic reticulum. However, we only detected a slight decrease in the root cytokinin levels. In contrast, a significant decrease of cytokinins occurred in the shoot. We suggest the roots as the main site of cytokinin biosynthesis in tobacco seedlings. Conversely, cytokinin levels in the shoot seem to depend largely on long-range transport of cytokinin ribosides from the root and their subsequent metabolic activation.

Cytokinins influence root gravitropism via differential regulation of auxin transporter expression and localization in Arabidopsis.

Redirection of intercellular auxin fluxes via relocalization of the PIN-FORMED 3 (PIN3) and PIN7 auxin efflux carriers has been suggested to be necessary for the root gravitropic response. Cytokinins have also been proposed to play a role in controlling root gravitropism, but conclusive evidence is lacking. We present a detailed study of the dynamics of root bending early after gravistimulation, which revealed a delayed gravitropic response in transgenic lines with depleted endogenous cytokinins (Pro35S:AtCKX) and cytokinin signaling mutants. Pro35S:AtCKX lines, as well as a cytokinin receptor mutant ahk3, showed aberrations in the auxin response distribution in columella cells consistent with defects in the auxin transport machinery. Using in vivo real-time imaging of PIN3-GFP and PIN7-GFP in AtCKX3 overexpression and ahk3 backgrounds, we observed wild-type-like relocalization of PIN proteins in the columella early after gravistimulation, with gravity-induced relocalization of PIN7 faster than that of PIN3. Nonetheless, the cellular distribution of PIN3 and PIN7 and expression of PIN7 and the auxin influx carrier AUX1 was affected in AtCKX overexpression lines. Based on the retained cytokinin sensitivity in pin3 pin4 pin7 mutant, we propose the AUX1-mediated auxin transport rather than columella-located PIN proteins as a target of endogenous cytokinins in the control of root gravitropism.

Short communication: Pasteurization as a means of inactivating staphylococcal enterotoxins A, B, and C in milk.

Our aim was to assess the effect of pasteurization temperature on inactivation of staphylococcal enterotoxins (SE). Milk samples were inoculated with log 4.38 to 5.18cfu/mL of 40 different Staphylococcus aureus strains having the ability to produce types A, B, or C SE and incubated at 37°C for 24h to develop SE. This incubation was followed by heat treatment for 15 s at 72, 85, and 92°C. Samples were analyzed for Staph. aureus count by plate method and, specifically, for SE presence. An enzyme-linked immunofluorescent assay on a MiniVIDAS analyzer (bioMérieux, Marcy l'Étoile, France) was used to detect SE, which were determined semiquantitatively based on test values. The Staph. aureus count in milk before pasteurization did not affect the amount of SE. Before pasteurization, SEB was detected in the lowest amount compared with other SE types. Staphylococcal enterotoxins were markedly reduced with pasteurization and inactivated at pasteurization temperatures to an extent depending on the amount in the sample before pasteurization. After pasteurization at 72°C, SE were detected in 87.5% of samples (35/40), after pasteurization at 85°C in 52.5% of samples (21/40), and after pasteurization at 92°C in 45.0% of samples (18/40). We determined that SE may still persist in milk even when Staph. aureus bacteria are inactivated through pasteurization. Although pasteurization may partially inactivate SE in milk, a key measure in the prevention of staphylococcal enterotoxicosis linked to pasteurized milk consumption is to avoid any cold chain disruption during milk production and processing.

The effect of cold chain disruption on the microbiological profile of chilled chicken meat.

This study evaluates the influence of inadequate transport conditions on the microbiological quality of chilled chicken meat packaged in plain and modified atmosphere packaging (MAP). The experiments simulated the temperature increase during sample transport to 8, 11, 14, 17, 20, and 25°C with exposure times of 1, 2, 3, and 4 h. Aerobic plate count (APC), psychrotrophic microorganisms count (PMC), ß-D-glucuronidase-positive Escherichia coli, and Salmonella spp. were evaluated immediately after the exposure to the elevated temperature (0 h), 3 h, and 24 h after the return to the temperature of ≤4°C. The upper acceptable limits for APC and PMC were set for each combination of investigated chicken meat and packaging type, taking also the initial bacterial condition into account. Chilled chicken breast samples in plain packaging exceeded the APC limits in 16 cases and PMC limits in 20 cases when exposed to temperatures of >4°C, while only 2 MAP samples exceeded APC limits and 8 samples PMC limits, respectively. In chicken legs, 8 samples in plain packaging exceeded the APC limits and 15 the PMC limits, while 12 samples in MAP exceeded the APC limits and 19 the PMC limits. In 402 samples (31.9%) in which the presence of E. coli was detected, its amount ranged from 1.70 to 3.65 log CFU.g-1. It was more commonly detected in chicken legs (255 of 630; 40.5%) than chicken breasts (147 of 630; 23.3%) but was not related to exposure temperature, exposure time, or time until examination. The presence of Salmonella spp. was not detected in any of the samples. Data acquired in the presented study will be used in the development of software helping the national supervisory authorities in the Czech Republic to evaluate whether inadequate transport of samples to analytical laboratories could have affected the microbiological profile of the sample.

Research Note: A mixture of Bacteroides spp. and other probiotic intestinal anaerobes reduces colonization by pathogenic E. coli strain O78:H4-ST117 in newly hatched chickens.

An experimental group of one-day-old chicken from a commercial hatchery was given a defined mixture of 7 gut anaerobes. The next day the chicks were inoculated by an APEC strain O78:H4-ST117 resistant to ciprofloxacin, alongside with the control group and monitored for 4 wk after the inoculation for the presence of the colonizing strains and ciprofloxacin-resistant E. coli. Significant reduction of colonization rates in the first 2 wk was recorded in the experimental group for the numbers of ciprofloxacin-resistant E. coli. The results show that colonization of chicken by defined anaerobic mixtures may provide a decisive protection during the critical period of the chicken intestinal microflora development.

Distribution of mercury in rainbow trout tissues at embryo-larval and juvenile stages.

The aims of the study were to determine total mercury concentrations in "rainbow trout Oncorhynchus mykiss (Walbaum)" at their embryo-larval and juvenile stages and to assess mercury concentration dynamics in individual tissues. Samples of rainbow trout were collected at two-month intervals over a period of 18 months (one stock production cycle) at the Velká Losenice trout farm. Feedstuff samples were collected at the same time and analyzed for mercury concentrations. Tissue mercury concentrations were determined in muscle, liver, and kidneys. Analyses were performed using the AMA 254 atomic absorption spectrophotometer. The lowest mercury concentration was found in 14-day-old embryos (hard roe), and the highest concentrations in muscle tissue, liver, and kidneys at the end of monitoring, that is, in rainbow trout aged 18 months. The amount of mercury in feedstuffs showed an increasing trend and ranged between 0.0126 and 0.0859 mg kg(-1). A significant effect (P < 0.001) of mercury intake on mercury concentrations in muscle tissue, liver, and kidneys was demonstrated. Muscle mercury concentrations in 18-month-old market-ready rainbow trout of 0.128 ± 0.048 mg kg(-1) met the criteria for fish meat hygiene.

Mercury content in fish from drinking-water reservoirs in the Morava River Basin (Czech Republic).

This study focused on the total mercury content in fish from seven drinking-water reservoirs located in the Morava River Basin: Bojkovice, Boskovice, Hubenov, Karolinka, Landstejn, Ludkovice and Nova Rise in the Czech Republic. A total of 308 fish were collected for the analysis. The content of total mercury was measured in the muscle tissue of bream, roach and perch using atomic absorption spectrometry and varied from 0.057±0.009 to 0.440 mg kg-1 in bream, from 0.030±0.005 to 0.393±0.138 mg kg-1 in roach and from 0.092±0.007 to 0.638±0.042 mg kg-1 in perch. The highest total mercury content was found in perch from Landstejn and the lowest was measured in roach from Ludkovice. A positive statistically significant relationship was found between fish weight and total mercury content in fish muscle for almost all species and all sampling sites, except for roach from Ludkovice. A total of 19 samples exceeded the maximum mercury level set by legislation on food contaminants-0.5 mg kg-1 in freshwater fish.

Research Note: Impact of applied thermal treatment on textural, and sensory properties and cooking loss of selected chicken and turkey cuts as affected by cooking technique.

The effect of various cooking methods (roasting, broiling, grilling, frying, and stewing) on cooking loss (CL) and textural and sensory properties of selected chicken (breast fillet, thigh, and thigh fillet) and turkey (breast fillet, thigh) cuts in relation to the applied apparatus was evaluated. Diverse results were recorded according to the method, the type of poultry meat, and the cut of poultry meat. Additionally, CL and shear force (SF) values in all examined samples were influenced by the culinary technique, the type of poultry meat, and the poultry meat cut. The lowest CL and shear SF values were reported when the samples were treated using a method with higher heating rates and/or temperatures and shorter cooking times. Additionally, lower values of CL and SF were obtained for chicken meat compared to turkey meat (thighs). In general, the applied culinary technique affected the sensory properties of the samples tested. High sensory scores were recorded for grilled chicken breast fillets and fried turkey breast fillets (irrespective of the applied apparatus). On the whole, it could be stated that culinary techniques at high temperature requiring shorter times (such as frying, grilling, and roasting) were evaluated to be more effective (in terms of CL and SF).

Cooking loss in retail beef cuts: The effect of muscle type, sex, ageing, pH, salt and cooking method.

The aim of the study was to compare cooking losses in beef (striploin and cube roll) during various cooking methods (grilling, roasting, broiling) in two different cooking devices (convection oven and clamshell grill). A total of 400 samples of meat were cooked (an internal temperature of more than 70 °C) and analysed. Cooking losses of slices of striploin ranged between 24.5% and 34.8%, with losses of 25.9-35.8% when whole cuts of meat were roasted. Losses fell within similar ranges (24.7-33.7%) for cube roll. Beef after 21 days of dry ageing showed generally lower cooking losses. Principal component analysis confirmed a strong negative correlation between cooking losses and salt content, saltiness and juiciness, which were positively correlated with one another. The majority of the sensory properties were positively correlated with pH, while their relationship with Warner-Bratzler (WB) test was negative. Roasting in a convection oven would seem to be the most suitable method of cooking striploin, followed by broiling in a convection oven, with grilling in last place. The highest score for the majority of sensory properties and the lowest WB values were recorded in meat roasted in a convection oven, particularly in the meat of bulls aged for 21 days.

Assessment of contamination of the Svitava and Svratka rivers in the Czech Republic using selected biochemical markers.

The aim of the present study is to assess aquatic ecosystem contamination using selected biochemical markers: cytochrome P450, ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), tripeptide glutathione, vitellogenin, and 11-ketotestosterone in chub (Leuciscus cephalus L.). Seven locations on the Svitava and Svratka rivers (in the Brno conurbation, Czech Republic) were assessed. The results were compared with the levels of the most important inductors of these biomarkers: organic pollutants hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), DDT and its metabolites, polychlorinated biphenyls, and polycyclic aromatic hydrocarbons in bottom sediment, fish muscle, and semipermeable membrane devices (SPMDs) and metals in sediment. The highest levels of pollutants were observed at sites situated downstream from Brno, especially at Modrice and Rajhradice. Significant positive correlations (p < 0.05) were found between EROD activity and HCH concentration in SPMDs, and also between GST and EROD activity with HCB concentration in muscle, after adjusting for age.

Using human hair as an indicator for exposure to mercury.

OBJECTIVES: Exposure to mercury, a risk factor for neuro-developmental toxicity, was evaluated in the Czech Republic by performing mercury determination using human hair as an indicator. METHODS: Hair samples from Czechs (n=311; 2-66 years old) were analyzed for mercury content. Total mercury was analysed by mercury analyzer AMA 254. RESULTS: The highest total mercury content found in sampled hair was 3.55 microg/g and the lowest content was 0.015 microg/g. No correlation was found between the mercury levels in the sampled hair and the subject's age, gender, and the amount of amalgam fillings. A total of 38 hair samples were analyzed for methylmercury content. CONCLUSION: The results show a positive correlation between the total mercury content in human hair and the consumption of marine and freshwater fish. Hair are a very good indicator of fish consumption.

Effect of metals, with special attention of Cd, content of the Svitava and Svratka rivers on levels of thiol compounds in fish liver and their use as biochemical markers.

OBJECTIVES: The aim of this study was to assess the effect of metals, with special attention of cadmium (Cd), content of the Svitava and Svratka rivers (Czech Republic) on levels of thiol compounds such as metallothioneins (MT), glutathione (GSH/GSSG), and cysteine (Cys) concentrations in chub (Leuciscus cephalus L.) liver and their use as biomarkers. DESIGN: Levels of thiol compounds in fish liver were determined by the differential pulse voltammetry Brdicka reaction. Cadmium content in L. cephalus liver was measured by atomic absorption spectometry (AAS) with electrothermal atomization. The contents of thiols and Cd were correlated with the Cd and metals content of river bottom sediments. RESULTS: Results were inconsistent with those of experimental studies. Correlations between liver Cd content and MT, Cys, GSSG were not significant in any locality (p > 0.05). Cd liver content was negatively (p< 0.05) correlated with GSH at one (Zidlochovice) of the seven tested sites. GSH and GSSG correlated negatively in five localities (p > 0.05). Correlations between Cd in sediment and Cys and GSSG (p > 0.05), and MT was negative (p< 0.05). The relationship between sum of metals value and MT was negative (p< 0.05), similarly with GSSG (p > 0.05). CONCLUSION: Many factors influence content of MT, GSH, and GSSG, and data for Cd were equivocal. Monitoring the aquatic ecosystem to identify metal content using thiol compounds in fish liver as biomarkers is complex. More relevant data including laboratory experiments will be required to assess most of the factors influencing thiols and their relevance to Cd contamination.

Fish biochemical markers as a tool for pollution assessment on the Svitava and Svratka rivers, Czech Republic.

OBJECTIVES: The study was designed to assess the pollution of the Svitava and Svratka rivers in and around the industrial city of Brno (Czech Republic) by persistent organic pollutants using selected biochemical markers in chub. DESIGN: Levels of selected biochemical markers were measured in liver and plasma samples of chub. The concentrations of persistent organic pollutants (POPs) were determined in bottom sediment, semi-permeable membrane devices (SPMDs) and muscle samples, and consequently used for correlation with biochemical markers. RESULTS: Significant alterations (p < 0.05) in some biochemical markers were observed and associated with combined exposure to pollutants. The highest levels of pollutants were found at sites situated downstream from Brno. The most widespread changes were identified in the function of phase I detoxifying enzymes. Significant positive correlations were observed in cytochrome P450 content and DDT concentration in the semi-permeable membrane device (p = 0.019, rs = 0.886), and between ethoxyresorufin-O-deethylase activity and content of DDT (p = 0.041, rs = 0.352) and polychlorinated biphenyls (p = 0.034, rs = 0.365) in muscle tissues of indicator fish. CONCLUSION: The results presented in our study indicate the highest contamination of sites situated downstream from Brno, where the intensive industrial and agricultural activities as well as domestic waste and sewage most probably comprise the main impact sources of the enhanced level of pollutants and some biochemical markers in fish.

Effect of cadmium chloride on metallothionein levels in carp.

Due to anthropogenic activities, heavy metals still represent a threat for various trophic levels. If aquatic animals are exposed to heavy metals, we can obviously observe considerable toxicity. It is well known that organisms treated with heavy metals synthesize low molecular mass compounds rich in cysteine. In this work the effects of cadmium chloride (2.5, 5, 7.5, 10 and 12.5 mg/L) on common carp (Cyprinus carpio) was investigated. We determined cadmium content in tissue of muscle, liver and kidney by atomic absorption spectrometry with electrothermal atomization and content of metallothionein (MT) in the same tissues by the Brdicka reaction. Electrochemical methods can be considered as suitable and sensitive tools for MT determination in carp tissues. Results of our study showed a gradually enhancing of cadmium content in muscle with time and dose of cadmium chloride in water. MT levels in liver reached both high levels (above 130 ng/g) in fish exposed to 2.5, 5 and 7.5 mg/L and low level (to 50 ng/g) in fish exposed to 10 and 12.5 mg/L of cadmium chloride. This finding confirms that the synthesis of metallothioneins and binding capacity of these proteins is restricted.

Leeches as Sensor-bioindicators of River Contamination by PCBs.

The aim of the study was to evaluate the use of leeches of the genus Erpobdella as a means of assessing polychlorinated biphenyl contamination of watercourses. The River Skalice, heavily contaminated with PCBs, was selected as a model. The source of contamination was a road gravel processing factory in Rozmitál pod Tremsínem from which an estimated 1 metric ton of PCBs leaked in 1986. Levels of PCB were measured in leeches collected between 1992 to 2003 from 11 sites covering about 50 km of the river (the first sampling site upstream to the source of contamination and 10 sites downstream). The PCB indicator congeners IUPA no. 28, 52, 101, 118, 138, 153, and 180 were measured. Levels were highest at the four sampling sites nearest the source of pollution. The highest values of PCB congeners were found in 1992. PCB content decreased from 1992 to 2003 and with distance from the source. The study indicated that leeches of the genus Erpobdella are a suitable bioindicator of contamination in the surface layer of river sediments.

When a river is affected by a reservoir: Trophic interactions and flexibility in feeding strategies of alpine bullhead (Cottus poecilopus) and European bullhead (Cottus gobio).

The alpine bullhead and European bullhead are benthic fish species protected by law in several European countries. One of the problems of European rivers is the ever-increasing number of reservoirs, which has adverse impacts on aquatic ecosystems. Feeding ecology of both species evaluated separately has been a subject of several studies, however, none of the studies paid attention to feeding interactions between them. Thus, the aims of the study were to evaluate trophic interactions of the two bullhead species and to assess their ability to cope with environmental disturbances caused by a reservoir. The study area consisted of three sites on the river Cierny Váh (Slovakia) that differed from each other by the intensity of pressures posed by the reservoir. The two species were found to coexist at two sites below the reservoir, because the reservoir affected their distribution along the river. For alpine bullhead, chironomid, hydropsychid and baetid larvae were the most important prey items, whereas the diet spectrum of European bullhead contained hydropsychid, chironomid larvae and detritus. Differences in diet composition were found among sampling sites. Temporal variations in the diet composition demonstrated that alpine bullhead is a more flexible feeder than European bullhead. Being predominantly food generalists, both species were able to shift their feeding strategies towards specialization when exposed to environmental disturbances at the most affected site. The reservoir increased the diet overlap between the two species, and thus changed their trophic interactions. Both alpine bullhead and European bullhead managed to cope with environmental disturbances caused by the reservoir, however, in lower population densities. Moreover, the reduced diversity of resources increased the potential for trophic competition between them.

Occurrence of Foodborne Agents at Food Service Facilities in the Czech Republic.

HIGHLIGHTS: Contamination of food service facilities in the Czech Republic by foodborne agents was determined. Bacillus cereus, Staphylococcus aureus, and Escherichia coli were detected in almost 50% of samples. The occurrence of B. cereus, S. aureus, and E. coli depended on the season of the year. Regular monitoring of food service facilities for agents of foodborne disease is necessary.