Relationships between pesticides, polychlorinated biphenyls, blood parameters and oxidative stress of white stork Ciconia ciconia chicks in Poland.

The white stork Ciconia ciconia is a bird species located at the top of the trophic pyramid in grassland and wetland ecosystems. This charismatic species is susceptible to pesticides and their environmental residues. In 2016, we collected blood samples from 114 white stork chicks across Western and Southern Poland. Chicks were sexed by molecular analysis and aged by development pattern. We studied the relationship between the concentration of pesticides (beta-HCH, heptachlor, aldrin, endrin, 4.4'-DDD, 4.4'-DDE and 4.4'-DDT) and of PCB in the chicks' blood with blood morphology and biochemistry parameters in the blood. The mean (± SD) values of concentrations of above detection level pesticides were: for (1) beta-HCH 4.139 ± 19.205; (2) 4.4'-DDE 9.254 ± 91.491 and additionally (3) PCB 16.135 ± 44.777 ppb. We found negative relationships between beta-HCH and oxidative stress enzyme activity in the blood, between beta-HCH and leukocyte concentration and between 4.4'-DDE and catalase activity. We also found a positive relationship between the concentration of pesticides in blood and the age of chicks. Interestingly, we found a higher concentration of PCB in the blood of male stork chicks than in female stork chicks. We provide more evidence that the presence of pesticides in the environment can be a strong stress factor, shaping the health status of birds.

The Aroma Composition of Korycinski Cheese Ripened in Different Temperatures.

As a regional product, Korycinski cheese is one of the most important cheeses in the Podlasie region of Poland. In this study, the influence of technological processes, such as ripening, on shaping the organoleptic characteristics of cheese was determined. Korycin-type cheeses are produced from cow's milk according to traditional technology. The ripening process lasted 466 h at 5 °C, 10 °C, and 15 °C. The aging temperature had a decisive influence on the number of esters and organic acids formed, which were analyzed by gas chromatography-mass spectrometry (GC/MS). The organoleptic properties of the cheeses were also related to the ripening temperature.

Could mycotoxigenic Fusarium sp. play a role in ulcerative dermal necrosis (UDN) of brown trout (Salmo trutta morpha trutta)?

Fusarium infections have been reported in aquatic animals, but are still poorly investigated in wild salmonids. The aim of the study was to determine the impact of the fungi and their toxins on the health status of brown trout (Salmo trutta morpha trutta) migrating from the Baltic Sea to the freshwater. Individuals from the wild brown trout population exhibiting ulcerative skin lesions were collected from the Slupia River in Poland and subjected to microbiological, histopathological, and hematological examinations, as well as toxicological analysis for a presence of mycotoxins. The results of microflora isolation from the brown trout skin samples revealed the presence of conditionally pathogenic bacteria and fungi classified by molecular techniques as Fusarium spp. Toxicological analysis allowed for detection of zearalenone (ZEN) in the liver, kidney, and gastrointestinal tract of the fish. In several cases, there was α-zearalenone (α-ZEL) identified at trace levels in the liver, as well as sterigmatocystin and enniatin B at low levels in the kidney and the liver. Histopathological examination revealed the presence of fungal hyphae disrupting the epidermis and penetrating into the necrotic dermis and hypodermis. The decreased values of the blood parameters, i.e., hemoglobin concentration (HGB), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and white blood cell count (WBC), were indicative of osmoregulation failure being a consequence of the skin damage. The results of the study provide new information regarding Fusarium sp. infection in brown trout and serve as the basis for further research on the potential impact of the fungi and their mycotoxins on the Baltic salmonid population, including their role in ulcerative dermal necrosis.

Do agricultural environments increase the reproductive success of White Stork Ciconia ciconia populations in South-Western Poland?

As populations of White Stork Ciconia ciconia have decreased at different rates in Europe, the specific environmental drivers that influence breeding success are a matter of controversy. Here we use body size, blood, and environmental data of a total of 1226 stork nestlings of different ages from a total of 363 nests in three different habitats (meadows, forest-edges, open agricultural landscapes) in S-W Poland to ask whether: 1) natural grassland environments increase the reproductive output in comparison with agricultural landscapes, 2) nestlings from agricultural landscapes differ in health status from nestlings in more natural habitats, 3) differences in environmental stress translate into respective differences in reproductive output and health status of nestlings. There was no significant difference in age corrected body mass and in the temporal increase in nestling mass between the study sites. Clutch sizes were highest and age corrected total clutch mass lowest at the meadow sites while agricultural and woodland sites did not significantly differ. Hemoglobin and red blood cell content were lowest and white blood cell and blood antioxidant concentrations were highest in the meadows indicating higher degrees of environmental stressors. These blood parameters varied strongly among study years. Our study does not confirm that agricultural landscapes are less suited for stork breeding success. We even find some indication of a better health status of nestlings in agricultural environments that might compensate smaller clutch sizes. Our data indicate that reproductive output is multifaceted. As we found some indication of a trade-off between clutch size and health status we argue that only multiple metrics of reproductive success are able to assess the long-term effect of habitat choice on fitness.

Relatedness of wildlife and livestock avian isolates of the nosocomial pathogen Acinetobacter baumannii to lineages spread in hospitals worldwide.

The natural habitats and potential reservoirs of the nosocomial pathogen Acinetobacter baumannii are poorly defined. Here, we put forth and tested the hypothesis of avian reservoirs of A. baumannii. We screened tracheal and rectal swab samples from livestock (chicken, geese) and wild birds (white stork nestlings) and isolated A. baumannii from 3% of sampled chicken (n = 220), 8% of geese (n = 40) and 25% of white stork nestlings (n = 661). Virulence of selected avian A. baumannii isolates was comparable to that of clinical isolates in the Galleria mellonella infection model. Whole genome sequencing revealed the close relationship of an antibiotic-susceptible chicken isolate from Germany with a multidrug-resistant human clinical isolate from China and additional linkages between livestock isolates and human clinical isolates related to international clonal lineages. Moreover, we identified stork isolates related to human clinical isolates from the United States. Multilocus sequence typing disclosed further kinship between avian and human isolates. Avian isolates do not form a distinct clade within the phylogeny of A. baumannii, instead they diverge into different lineages. Further, we provide evidence that A. baumannii is constantly present in the habitats occupied by storks. Collectively, our study suggests A. baumannii could be a zoonotic organism that may disseminate into livestock.

Sex ratio of White Stork Ciconia ciconia in different environments of Poland.

The aim of this study was to analyze the variation in sex ratio of White Stork Ciconia ciconia chicks from differentiated Poland environments. We took under a consideration the impact of Cd and Pb for establish differences among sex ratio in chicks. We also study multiplex PCR employment for establish gender considerations. We collected blood samples via venipuncture of brachial vein of chicks during 2006-2008 breeding seasons at the Odra meadows (SW-Poland; control), which were compared with those from suburbs (SW-Poland), and from copper smelter (S-Poland; polluted) and from swamps near Baltic Sea. We found differences among sex ratio in White Stork chicks from types of environment. Male participation in sex structure is importantly higher in each type of environment excluded suburban areas. Differences in White Stork sex ratio according to the degree of environmental degradation expressed by Cd and Pb and sex-environment-metal interactions testify about the impact of these metals upon sex ratios in storks. Simultaneously, as a result of multiplex PCR, 18S ribosome gene, which served as internal control of PCR, was amplified in male and female storks. It means that it is possible to use primers designed for chicken in order to replicate this fragment of genome in White Stork. Moreover, the use of Oriental White Stork Ciconia boyciana W- chromosome specific primers makes it possible to determine the sex of C. ciconia chicks. Many factors make sex ratio of White Stork changes in subsequent breeding seasons, which depend significantly on specific environmental parameters that shape individual detailed defense mechanisms.

Corynebacterium pelargi sp. nov., isolated from the trachea of white stork nestlings.

A Gram-stain-positive, pleomorphic, oxidase-negative, non-motile isolate from the trachea of a white stork from Poland, designated strain 136/3(T), was subjected to a comprehensive taxonomic investigation. A comparative analysis of the 16S rRNA gene sequence showed highest similarities to Corynebacterium mustelae , Corynebacterium pseudotuberculosis , Corynebacterium vitaeruminis and Corynebacterium ulcerans (96.0-96.3%). The quinone system consisted of major amounts of MK-8(H2), minor amounts of MK-9(H2) and traces of MK-8 and MK-9. The polar lipid profile of strain 136/3(T) contained phosphatidylinositol and phosphatidylinositol-mannoside as major lipids and phosphatidylglycerol and an acidic glycolipid in moderate amounts. In addition small amounts of diphosphatidylglycerol, a phospholipid, an aminolipid and two lipids of unknown group affiliation were found. The polyamine pattern was composed of the major components spermidine and spermine. Putrescine, 1,3-diaminopropane, cadaverine, sym-homospermidine and tyramine were found in minor or trace amounts. The diamino acid of the peptidoglycan was meso-diaminopimelic acid. In the fatty acid profile straight-chain, saturated and mono-unsaturated fatty acids predominated (C(18 : 1)ω9c, C(16 : 1)ω7c, C16 : 0, C(18  : 0)). Corynemycolic acids were detected. Physiological traits as well as unique traits of the polar lipid profile and the fatty acid pattern distinguished strain 136/3(T) from the most closely related species. All these results indicate that strain 136/3(T) represents a novel species of the genus Corynebacterium for which we propose the name Corynebacterium pelargi sp. nov. The type strain is 136/3(T) ( =CIP 110778(T) =CCM 8517(T) =LMG 28174(T)).

Prevalence, virulence, and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli in white stork Ciconia ciconia in Poland.

The aim of this study was to investigate the role of white stork Ciconia ciconia as a potential reservoir of Campylobacter spp. Antimicrobial resistance and the presence of putative virulence genes of the isolates were also examined. A total of 398 white stork chicks sampled in Western Poland in habitats with high density of breeding were examined. Rectal swabs were collected during breeding season 2009-2012 from storks developing in a relatively pure environment (Odra meadows), in polluted areas (a copper mining-smelting complex), and in suburbs. Of the anal swabs collected, 7.6% were positive for Campylobacter among chicks (5.3% samples positive for C. jejuni and 2.3% samples positive for C. coli). Samples from polluted areas had the highest prevalence of Campylobacter (12.2%). The prevalence of resistance among C. jejuni and C. coli isolates from young storks was as follows: to ciprofloxacin (52.4%, 44.4%), and to tetracycline (19%, 77.8%). All of the analyzed isolates were susceptible to macrolides. The resistance to both classes of antibiotics was found in the 23.3% of Campylobacter spp. All Campylobacter spp. isolates had cadF gene and flaA gene responsible for adherence and motility. CdtB gene associated with toxin production was present in 88.9% of C. coli isolates and 57.1% of C. jejuni isolates. The iam marker was found more often in C. coli strains (55.6%) compared to C. jejuni isolates (42.9%). Our results confirm the prevalence of Campylobacter spp. in the white stork in natural conditions and, because it lives in open farmlands with access to marshy wetlands, the environmental sources such as water reservoirs and soil-water can be contaminated from white stork feces and the pathogens can be widely disseminated. We can thus conclude that Campylobacter spp. may easily be transmitted to waterfowl, other birds, and humans via its environmental sources and/or by immediate contact.

[The susceptibility of Proteus mirabilis strains isolated from white stork (Ciconia ciconia)]. / Lekowrazliwosc szczepów Proteus mirabilis izolowanych od bociana bialego (Ciconia ciconia).

Proteus sp. rods are ubiquitous bacteria, widespread in the environment and classified also as opportunistic human pathogens. The aim of our study was to evaluate susceptibility of Proteus mirabilis strains isolated from white stork (Ciconia ciconia) regarding as his natural bacterial flora, compare and discuss their results with data obtained from scientific literature for clinical strains of the same species. Susceptibility of 59 P. mirabilis strains was estimated for 27 antimicrobials using disc-diffusion method and the ability to produce extended spectrum beta-lactamases was evaluated by double disc synergy test. Environmental P. mirabilis strains isolated from white stork were assessed as more susceptible to most of the examined antimicrobials and production of extended spectrum beta-lactamases was not noted amongst them.

Blood chemistry in white stork Ciconia ciconia chicks varies by sex and age.

Little is known on how blood biochemistry differs among avian chicks, especially in sexually monomorphic species. In this study we sampled blood chemistry of 342 white stork Ciconia ciconia chicks from nests in western Poland during four years (2005-2008). Special attention was paid to the effect of chick age and sex on blood biochemistry. Since white stork is a monomorphic species, the sex of chicks was established by a molecular technique. Nine blood biochemical parameters were studied: total protein concentration, urea, uric acid, triglycerides, total cholesterol, high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). There were sexual differences in total protein, uric acid, cholesterol, HDL and AST. However, total protein and uric acid only differed significantly between sexes if an age effect was included as a covariate in the analysis. Triglycerides decreased significantly, and AST, increased significantly with chick age. We confirm that blood biochemistry varies with chick age, but we also found significant differences between the sexes. Therefore, to understand changes in blood parameters, and to establish reference ranges useful in captive rearing of this endangered species, establishing gender may be important, even in very young individuals.

Ecophysiological determinations of antioxidant enzymes and lipoperoxidation in the blood of White Stork Ciconia ciconia from Poland.

The aim of this study was to investigate the activity of antioxidant enzymes in the blood of White Stork Ciconia ciconia chicks (aged 19-54 days) in Poland in 2006. We took under consideration superoxide dismutase (SOD), catalase (CAT), ceruloplasmine (CP), glutathione peroxidase (GPx), glutathione reductase (GR) and the content of thiobarbituric acid-reactive substances (malondialdehyde) in polluted (copper manufacture), suburban areas, at the Odra meadows, and at swamps near Baltic Sea in the Pomeranian region. We examined the levels of Na, K, Ca, Mg, Fe, Zn, Cu, Mn, Co, Cd, and Pb and compared ecophysiological determinations for developing storks. Blood samples of wing venous were collected from 91 chicks from 33 nests. The degree of activity of antioxidant enzymes studied has been different in White Stork chicks' blood from Poland regions, as a rule. We have stated a relatively high level of CAT, GPx, SOD, and GR activity and malondialdehyde (MDA) content in chicks from polluted areas. However, relative value for GR in storks from Odra meadows was considerably higher (about 112 nmol NADPH(2)/min ml) than those in chicks from other environments (56-84 nmol on average). Relatively high levels of CAT, CP, and GPx (2.7 mkM/min l, 22.2 mg/l, and 3.8 nmol GSH/min ml, respectively) were also stated in chicks nested in swamps near Baltic Sea. Simultaneously, we have stated differences (p<0.02-p<0.001) in the level of elements (besides Ca) in blood of young storks from the studied areas. We found a high level of toxic metals, e.g. Cd, either from swamps near Baltic Sea (2.7 mg/kg) or from Glogów smelter (2.2mg/kg), whilst Pb concentration was high in chicks from Glogów (7.2 mg/kg). Cd and Pb levels in blood of chicks were different in individuals from each region (p<0.001). Birds from a smelter have the highest level of these elements, whereas the lowest one was stated in chicks from Odra meadows (Cd: 1.45, Pb: 0.84 mg/kg). Thus, Cd could be a useful marker of response for polluted stress. We also observed a relatively high level of Mg in chicks from both Pomeranian (7000 mg/kg) and polluted (about 6000 mg/kg) areas. Potassium, zinc, and cobalt levels were highest in chicks from suburbs (4.65, 10.1, and 2.7 mg/kg, respectively) and polluted regions (3.8, 9.7, and 5.6 mg/kg, respectively), whilst Cu and Mn were highest in those from polluted (10.9 and 47.6 mg/kg, respectively) and Pomeranian regions (11 and 42.2 mg/kg, respectively). Concentrations of Na, K, and Ca in chicks from Glogów smelter (143.2, 3.8, and 115.9 mg/kg, respectively) were often similar to those from Odra meadows (147.8, 3.6, and 112.5 mg/kg, respectively). This was probably due to a similar degree of homeostatic regulations of an organism. The levels of Mg, Fe, Zn, and Cu were often different (p<0.02-p<0.001) in the blood of White Stork chicks from the studied areas. Co, Pb, and Cd levels were higher (p<0.001) in chicks from Glogów smelter than in those from Odra meadows. It is evidence for importance of anthropopression, which influenced the course of biogeochemical processes and the bioaccumulation of toxic metals locally. This takes place also in chicks from swamps near Baltic Sea, in which the level of Cd was high (2.7 mg/kg); so we can state the high intensity of intoxication in this region. We can conclude that the use of hematological research assesses the condition of birds and might give a positive association with miscellaneous environmental loads. The high concentration of toxic heavy metals involved greater intensity of antioxidant enzymes' activity. Environmental intoxication causes an increase of lipoperoxidation intensity in growing chicks and changes the response of their immunological system.

The impact of element-element interactions on antioxidant enzymatic activity in the blood of white stork (Ciconia ciconia) chicks.

The aim of this work was to determine interrelationships among macroelements Na, K, Ca, Mg, and Fe, microelements Zn, Cu, Mn, and Co, and toxic heavy metals Pb and Cd in the blood of white stork Ciconia ciconia, during postnatal development, in different Polish environments, and their impact on the activity of antioxidant enzymes. We considered the content of thiobarbituric acid-reactive substances (TBARSs), i.e., malondialdehyde (MDA), and activity of superoxide dismutase (SOD), catalase (CAT), ceruloplasmine (CP), glutathione peroxidase (GPx), and glutathione reductase (GR). Blood samples were collected from storks developing at Odra meadows (Klopot; southwestern Poland). They were compared with blood of chicks from several suburban sites located 20 km away from Zielona Góra (0.1 million inhabitants; southwestern Poland) and near Glogów, where a copper smelter is situated. We also conducted research in the Pomeranian region (Cecenowo; northern Poland). We collected blood samples via venipuncture of the brachial vein of chicks in 2005-2007. They were retrieved from the nest and placed in individual ventilated cotton sacks. The blood was collected using a 5-ml syringe washed with ethylenediaminetetraacetic acid (EDTA). We found significant interactions between macro- and microelements and enzymatic activity and TBARS products. We noticed the predominance of Cd and Pb participation in element-enzyme interactions. Simultaneously, we found interrelationships between cadmium and Na, K, Ca, Mg, and Fe and the activity of antioxidant enzymes SOD, CAT, CP, GR, and TBARS products in the blood of white stork chicks. In the case of lead these relationships were not numerous and they were significant for Ca, Mg, Cu, Mn, and Co. Correlations with enzymes were significant for Pb-CAT and Pb-TBARS. We noted that activities of most enzymes (SOD, CAT, CP, GR) and TBARS products are determined by their interactions with physiological elements Na, Ca, Mg, Fe, and Zn and toxic heavy metals. White stork chicks ranged in age from 17 to 59 days. Concentrations of elements in the blood were age related. Among enzymes, only SOD, CAT, and GPx were age related. Young storks differed in the case of element concentration (except for Ca, Zn, and Cd) and enzymatic activity. We found that significant element-element interaction/enzyme activity predominated in the case of physiological elements and toxic metals, which we explain by the intensive and prevailing access of toxic metals in redox reactions. This causes changes in the priority of these metals, reflected by their influence on the enzymatic activity of antioxidant enzymes. The content of Cd and Pb in blood of young storks from different regions tends to affect the lipid peroxidation process negatively. However, in many cases we observed an increase in enzymatic activity with an increase in heavy metals. This indicates the changes in oxidative stress intensity in chicks in response to environmental differentiation. The increase in lipoperoxidation modifies antioxidant enzyme activity and causes changes in SOD, CAT, CP, GPx, and GR activity in chicks from various regions, principally increases in enzyme activity in chicks from polluted environments and suburbs. We suggest that the source of heavy metals in chicks' blood might be used as a biological test system of adaptation to oxidative stress. We also report that a high level of heavy metals is accompanied by increased lipid peroxidation. Thus young storks are probably significantly susceptible to environmental conditions. They demonstrated initiation of lipoperoxidation and oxidative modification of proteins that coincide with chemical elements, as a possible antioxidant defense system.

Melatonin and thyroxine response to pollution in white stork nestlings (Ciconia ciconia): aspects of rhythmicity and age.

There is growing evidence that ubiquitous environmental contaminants may interfere with vertebrate endocrine systems. The selected endocrine biomarkers are used to indicate the condition of free-ranging populations of wildlife, including avian species. The aim of this study was to determine the impact of environment quality on serum thyroxine (T4) and melatonin (Mel) in white stork nestlings (Ciconia ciconia) living in different locations: small villages in natural areas surrounded by forests and crop fields, near the city and near the copper smelter. We extended our analyses to examine the hormones' day-night changes in conjunction with chicks' age. Total serum T4 and Mel was measured by RIA. T4 level, as a decisive measure of thyroid hormone productivity, was significantly lower in the nestlings exposed to pollutants from the copper smelter. Mel, as a well-known scavenger of free radicals, was elevated in the nestlings in the area near the copper smelter. This study indicates that alteration in T4 and Mel levels could be a useful marker of exposure of nestling wild storks to different toxic substances in field studies. Mel is postulated to be a susceptible defensive molecule as a protective mechanism for organisms.