Peroxidation and unsaturation indices as potential biomarkers of multifarious zinc and copper micro-supplementation in Helix pomatia L.

The work is a continuation of two previous studies in which biomarker fatty acids (12 of 56 FA pools) were analysed in Helix pomatia L. after heterogeneous micro-supplementation of Zn and Cu (administered in five micro-doses in the form of salts and EDTA and lysine chelates). This time, peroxidation (PI) and unsaturation coefficients (UI) as biomarker were analysed. These indices were calculated based on the FA profile in the foot and hepatopancreas of snails. The correlation of frequently used oxidation status indicators of organisms (catalase - CAT, glutathione peroxidase - GPx, selenium-dependent peroxidase - se-GPx, superoxide dismutase - SOD, glutathione transferase - GST, glutathione reductase - GR, glutathione - GSH, carbonyl protein - CP, thiobarbituric acid reactive substances - TBARS) with the rarely used UI and PI ratios was analysed. It was found that the 12-week micro-exposure to Zn and Cu did not inhibit but rather stimulated antioxidative defence at a sufficient level to increase the values of peroxidation/unsaturation indices in comparison to the control groups. Induction of an opposite process to oxidation of fatty acids was demonstrated. Maximum activities and amounts of antioxidants as well as minima of protein and lipid decomposition were recorded in groups supplemented with 0.75mg/l Zn and 1.0mg/l Cu. The possibility of a direct use of fatty acids as well as peroxidation/unsaturation indices as sensitive and reproducible biomarkers of exposure and oxidative physiological status in snails was confirmed.

Changes in fatty acid metabolism induced by varied micro-supplementation with zinc in snails Helix pomatia (Gastropoda Pulmonata).

We analyzed the changes in the profile of fatty acids (FA) in the foot tissues and hepatopancreas (HP) of snails Helix pomatia exposed to five microdoses of zinc (0.1, 0.25, 0.5, 0.75, or 1mg/l) administered in the form of a pure salt solution and in the form of EDTA and lysine chelates. Selection from a pool of 56 fatty acids analyzed in snail tissues yielded a set of 12 biomarker acids undergoing significant changes in contact with toxic substances. The selection criteria included the greatest percentage among the FA profile and their significant role in physiological processes. The proposed palette of acids of the biomarker FAs comprised C16:0; C18:0; C23:0; C18:1 n-9; C20:1 n-9; C18:2 n-6; C18:3 n-3; C20:2; C20:4 n-6; C20:5 n-3; C22:4 n-6; and C22:5 n-3, and saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs), determined separately in the foot tissues and hepatopancreas. The significant (p=0.01) influence of the dose as well as the source of the zinc on its' concentration in the tissues and on changes in the fatty acid profiles. Among the three zinc forms administered to the snails, the highest bioaccumulation of zinc in both tissues was noted in the group receiving the Zn-EDTA chelate. The content of PUFAs increased as the supplementation with zinc increased up to 0.75mg/l, but at 1mg/l, the share of these FAs began to decrease. This trend was observed in both analyzed tissue types - foot and hepatopancreas. The dose of 1mg Zn/l might be considered as a threshold dose above which the saturation of FAs increases. The results proved that determination of FA profile in snails can be used in ecotoxicological research as a reliable test of the effect of trace doses of stressors. The micro-supplementation of the mollusks diet with zinc is an example of a non-routine approach to issues connected with both diet and toxicology.

Effect of Raw Chickpea in the Broiler Chicken Diet on Intestinal Histomorphology and Intestinal Microbial Populations.

The aim of the study was to determine the effect of partial replacement of SBM protein with CPR-derived protein in the broiler rearing period from 22 to 42 days of age on the intestinal histomorphology and the composition of the intestinal microbial population. Male broiler chicks aged 1 day were assigned to two groups with different nutrition schemes (n = 100 in each, 5 cages of 20 chicks in each). The chickens were reared for 42 days. All birds were fed isonitrogenous and isoenergetic diets: starter (1 to 21 d), grower (22 to 35 d), and finisher (36 to 42 d). From rearing day 22, different diets were provided to the birds: the SBM group received feed with 100% soybean meal protein, and the diet of the CPR group the protein originated from soybean meal was replaced by 50% chickpea protein. The study results indicated a significant impact of the inclusion of CPR in the diet on the basic intestinal structures (thickness of myenteron: submucosa, jejunum and duodenum mucosa, and jejunum transversal lamina). The addition of CPR led to shortening of intestinal villi, an increase in villus thickness, and reduced intestine absorptive surface in the duodenum and jejunum. The CPR group exhibited a significantly lower villus length-to-crypt depth ratio in the jejunum (p < 0.001). The inclusion of chickpeas in the diet increased the total count of mesophilic bacteria and coliforms in the intestinal contents (p < 0.05). In summary, it has been demonstrated that the inclusion of CPR in the diet induced considerable disturbances in metabolism and intestinal structure. Although CPR is a cheap protein source, its use in poultry diet does not ensure development of the intestinal structure comparable to that in the case of an SBM-only diet.

Effect of inclusion of micronized camelina, sunflower, and flax seeds in the broiler chicken diet on performance productivity, nutrient utilization, and intestinal microbial populations.

The aim of the study was to evaluate the effect of inclusion of micronized full-fat camelina, flax, or sunflower seeds in the diet for broiler chickens on the performance productivity, nutrient utilization, and composition of intestinal microbial populations and to assess the possibility of modification of the resistance of isolated bacteria to chemotherapeutic agents with different mechanisms of action. The use of micronized oilseeds improved the broiler chicken body weight (P = 0.035) and the FCR value (P = 0.045) in the final rearing stage by enhancement of the utilization of total protein and organic matter. Lactobacillus-Enterococcus spp., Bifidobacterium spp., Escherichia coli, and Salmonella spp. were isolated from small intestinal contents, and Enterobacteriaceae taxa were detected in the cecum and cloaca of the broiler chickens. The addition of micronized camelina seeds (CAM.IR) contributed to an increase in the Bifidobacterium counts in the small intestine, compared with the control treatment (P < 0.050). Escherichia coli bacteria were not isolated only in the CAM.IR treatment. Nitrofurantoin and chloramphenicol were the most effective agents against the isolates from the cecum and cloaca in all oilseed treatments, whereas streptomycin exhibited the lowest efficacy. In the CAM.IR and micronized sunflower seed (SUN.IR) treatments, there were higher counts of trimethoprim/sulfamethoxazole-resistant Enterobacteriaceae strains than in the control and micronized flax seed (FLA.IR) treatments (P < 0.05). There was a difference between strains isolated from the cecum and cloaca only in the FLA.IR treatment, i.e., increased tetracycline sensitivity was exhibited by strains isolated from the cloaca (13% vs. 50%), also in comparison with the control treatments (P = 0.054). In comparison with the CAM.IR and control treatments, reduced numbers of multi-resistant strains were found in the cloaca isolates from the for FLA.IR and SUN.IR variants. Micronized camelina, flax, and sunflower seeds can be used as part of an effective nutritional strategy focused on optimization of the efficiency of rearing broiler chickens, as they positively modify intestinal microbial populations and increase bacterial sensitivity to the analyzed chemotherapeutic agents.

Peroxidation and unsaturation coefficients as biomarkers of environmental micro-exposure to molluscicides in Helix pomatia L.

This study was a follow-up of a previous study that investigated a set of selected fatty acids (FAs; 12 of 56 pools) in Helix pomatia L. as biomarkers of chemical stress induced by applying micro-doses of molluscicides. Here, the potential of rarely used peroxidation (PI) and unsaturation (UI) coefficients were tested as biomarkers. These indices were calculated based on the FA profiles of foot and hepatopancreas tissues of H. pomatia L. Snails were treated with three molluscicides: metaldehyde, methiocarb, and potassium chloride, in three doses each (5, 10, or 15 µl, 0.01% w/v concentration), for 12 weeks, under laboratory conditions. Correlations were evaluated between frequently used oxidation status indicators (catalase, glutathione peroxidase, selenium-dependent peroxidase, superoxide dismutase, glutathione transferase, glutathione reductase, glutathione, carbonyl protein, and thiobarbituric acid reactive substances, in the form of MDA) and UI and PI ratios. These results confirmed that fatty acids could be directly used as biomarkers of exposure and oxidative physiological status in snails. Moreover, the UI and PI, calculated based on FAs, clearly reflected the current oxidation status in snails. These indices changed with the application of micro-doses of molluscicides. In conclusion, these indices could serve as sensitive biomarkers of chemical stress in snails.

Selected fatty acids as biomarkers of exposure to microdoses of molluscicides in snails Helix pomatia (Gastropoda Pulmonata).

Two stages of selection from a pool of 56 fatty acids analyzed in Helix pomatia yielded a set of 12 biomarker acids undergoing significant changes in contact with three microdoses of toxic substances, i.e. three molluscicides containing metaldehyde, methiocarb, and potassium chloride (PC). The proposed palette of acids, including saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA), determined separately in the foot tissues and hepatopancreas of Gastropoda, can be used in ecotoxicological research as a reliable test of the effect of trace doses of stressors. The final set of the biomarker FA comprised C16:0; C18:0; C23:0; C18:1 n-9; C20:1 n-9; C18:2 n-6; C18:3 n-3; C20:2; C20:4 n-6; C20:5 n-3; C22:4 n-6; and C22:5 n-3. A clear physiological response manifested as changes in the content of fatty acids (FA) was observed in the snails even in the case of the lowest doses of the pollutants. All experimental factors analyzed, i.e., the dose (5, 10, or 15 µl 0.01% w/v concentration) and the type of preparation (metaldehyde, methiocarb or PC), had a significant (p ≤ 0.01) impact on the FA composition of the foot and hepatopancreas. Limitation of the analysis to a narrow pool of reactive FA meets the requirements of parameters of biomarkers of exposure and facilitates and accelerates visualization of the bioindicator organism's response to the presence of the stressor in the environment.

Tawny owl (Strix aluco) as a potential transmitter of Enterobacteriaceae epidemiologically relevant for forest service workers, nature protection service and ornithologists.

Established taxa within the Enterobacteriaceae wereisolated from cloacal swabs of Strix aluco chicks in nest boxes located at five research sites. ChromID ESBL medium (bioMerieux) was used to select a pool of Enterobacteriaceae strains producing extended-spectrum beta-lactamases. Drug sensitivity of the chosen strains was determined from the full pool of Enterobacteriaceae to 6 chemotherapeutics of different mechanisms of action. The study evaluated the sensitivity of ESBL-synthesizing isolates to substances belonging to penicillins, cephalosporins, cephamycins, clavams, carbapenems and monobactams. Analysis of the results indicated a potential role of Strix aluco in the dissemination of epidemiologically-relevant Enterobacteriaceae, and, importantly, pose health risks to forest service workers, nature protection service and ornithologists. The results can also serve as the basis for further environmental studies.

[Antimicrobial susceptibility and phage types of Salmonella enteritidis strains isolated in the Lublin area]. / Lekowrazliwosc oraz typy bakteriofagowe szczepów Salmonella enteritidis izolowanych na terenie makroregionu lubelskiego.

The study aimed at specifying the value of two traditional methods of typing, using the collection of 241 strains of Salmonella Enteritidis isolated from people and animals in the Lublin area in 1994-1995 from the occasional cases of infections. There were 8 phage types identified among the examined strains. Phage types PT 6 (40.24% of strains) and PT 7 (29.46%) were the most numerous ones. Salmonella Enteritidis was numbered among 38 profiles on the basis of the analysis of resistance to 15 antimicrobial agents. It was found that nitrofurantoin had the lowest efficacy in vitro in relation to the examined collection of Salmonella isolates. High percentage of strains were characterized by lack of susceptibility to streptomycin, neomycin, cefixime, tetracycline and canamycine. While analyzing the profiles of resistance to chemiotherapeutics and the expression of phage type of the examined strains it was observed that only 14.1% of all strains showing resistance to nitrofurantoin represented at the same time phage type PT 6, whereas 10.8% of strains belonging to the same resistance profile indicated PT 7 phage type. In correlated analysis of resistance to chemiotherapeutics and phage type expression of the examined collection of 241 isolates of Salmonella Enteritidis, 18.3% of strains showed distinct and unique set of these two features, which can be used in epidemiological investigations for the preliminary characterization of the bacterial population.