The effect of Limosilactobacillus fermentum 2i3 and 0.6% addition of humic substances on production parameters and the immune system of broilers.

The widespread use of antibiotics in the poultry industry as growth promoters has led to the emergence of bacterial resistance, which poses a significant health risk to humans and animals. Substances of natural origin, such as probiotic bacteria and humic substances, can be a promising solution. The aim of this experiment was to study the effect of the administration of a probiotic strain of Limosilactobacillus fermentum 2i3 and/or a new formula of humic substances specifically designed for detoxification on the production parameters, including gene expression of myogenic growth factors and selected parameters of the immune response. We found that production parameters such as feed conversion ratio and weekly weight gain, as well as gene expression of mucin-2 and immunoglobulin A, were positively influenced mainly by the administration of L. fermentum 2i3. Similarly, the percentage of active phagocytes and their absorption capacity as well as the proportions of CD8+ and CD4+CD8+ T-lymphocyte subpopulations were significantly increased. The addition of humic substances, either alone or in combination with probiotics, significantly reduced the aforementioned parameters compared to the control. On the other hand, the relative gene expression for all myogenic growth factors was the highest in the humic group alone. Based on the results obtained, we can confirm the immunostimulating effect of L. fermentum 2i3 administered in drinking water, which also had an impact on important production parameters of broiler meat. On the other hand, in the combined group there was no expected potentiation of the positive effects on the observed parameters.

Effect of fungal solid-state fermented product enriched with gamma-linolenic acid and ß-carotene on blood biochemistry and immunology of broiler chickens.

The aim of this study was to investigate the effect of the addition of fungal solid-state fer-mented product (FP) enriched with gamma-linolenic acid (GLA) and ß-carotene to feed on the haematological and immunological parameters of broiler chickens. Eighty 1-day-old COBB 500 broiler chickens were divided into two groups. The control group was fed with basic diets and chickens of the experimental group received 10% addition of FP, while the amount of basic diet was reduced. FP was produced during a solid-state fermentation (SSF) process using Umbellopsis isabellina CCF2412 as a producer of GLA and ß-carotene. After 38 days of feeding, blood sam-ples were collected and analyzed. Lower total and LDL-cholesterol values were measured in blood samples of the experimental animals (p⟨0.05). However, the triacylglycerol content was higher in the experimental group (p⟨0.05). Significantly higher levels of hematocrit and hemo-globin, and lower eosinophil and basophil content in the experimental group were recorded (p⟨0.05). The experimental group showed higher numbers of B lymphocytes and greater phago-cytic capacity (p⟨0.05). The results indicate that a fermented product produced by SSF, using the fungal strain Umbellopsis isabellina, is a good source of GLA and ß-carotene, which can influence the biochemical, hematological and immunological parameters of broiler chickens.

Effect of fungal gamma-linolenic acid and beta-carotene containing prefermented feed on immunity and gut of broiler chicken.

Gamma-linolenic acid (GLA) is a fatty acid from the ω-6 family. It is able to deliver a wide range of health benefits arising from its anti-inflammatory effects. An insufficient supply of GLA from agricultural and animal sources resulted in the development of a fermentation technique using lower filamentous fungi, which have the ability to accumulate high concentrations of GLA and beta-carotene during solid-state fermentation of cereals. The goal of this study was to observe the influence of the addition of prefermented cereal product, containing high amounts of GLA and beta-carotene, into the feed of broiler chickens on their immune status, and also the number of lactic acid bacteria and enterobacteria in gut content, which has never been studied before. Immunostimulation in the GLA group was manifested by a significant increase in the oxidative burst of phagocytes, CD4+CD8- lymphocytes in blood, and the CD4: CD8 ratio. Upregulation of gene expression for IgA in the GLA group indicates that the B-lymphocytes were stimulated at a local gut level. In the caecum, increased mRNA expression for mucin-2 and insulin-like growth factor was observed in the GLA group, which could contribute mainly to the protection of the intestinal mucosa and to better growth and regeneration of skeletal muscles. Improved immune activation and protection of the intestinal mucosa were subsequently reflected in a change of the microbial composition in gut contents; a significant reduction of enterobacteria occurred after GLA administration. We can conclude that prefermented cereals containing fungal GLA and beta-carotene represent a low-cost supplement for broiler diet having a beneficial health effect.

Effect of dietary microbially produced gamma-linolenic acid and plant extracts on enzymatic and non-enzymatic antioxidants in various broiler chicken organs.

Plant extracts and fungal fermented feed with gamma-linolenic acid-rich microbial oils are perspective additives for use in animal nutrition as appetite and digestion stimulants, stimulants of physiological functions, for the prevention and treatment for certain pathological conditions, and as antioxidants. The activity of antioxidant enzymes and the level of reduced glutathione were measured in the plasma and in liver, heart and kidney mitochondria after 42 days of feeding broiler chickens both regular and combination diets. These were selected based on our previous experience. The administration of agrimony and gamma-linolenic acid resulted in a significant decrease in superoxide dismutase activity in all four bodies in contrast to plant extracts. We conclude that the decrease in activity is due to decreased production, and hence dismutation, of superoxide radicals to peroxides followed by lower activity of glutathione peroxidase, which was not seen in the case of only plant extract administration. Generally, higher glutathione reductase activity would be in response to increased demands on reduced glutathione as a cofactor for the reaction catalysed by glutathione peroxidase and the utilization of glutathione itself. However, measured levels of reduced glutathione showed no change. The results argue against any oxidative stress conditions. The application of agrimony extract appears to be suitable for the antioxidant effect against peroxidation of gamma-linolenic acid. As the efficacy of measuring the effects of diets on the oxidative stability of meat caused by selected antioxidant enzymes is rather low, additional data from the experiment will be processed to clearly assess the influence of this combination of diets.

The effect of supplementation of clove and agrimony or clove and lemon balm on growth performance, antioxidant status and selected indices of lipid profile of broiler chickens.

The study investigated the effects of diet supplementation with 1% clove flower buds powder combined with either 0.2% lemon balm extract or 0.2% agrimony extract (each of the two pulverized extracts supplied through drinking water) on body weight of broilers, total feed intake, feed conversion ratio and the carcass yield, activity of superoxide dismutase (SOD, EC 1.15.1.1) and glutathione peroxidase (GSH-Px, EC 1.11.1.9) in blood, concentration of sulfhydryl (-SH) groups, malondialdehyde (MDA), vitamin A and E, low-density lipoproteins in the blood plasma, serum cholesterol, total lipids, triglycerides and high-density lipoproteins in broiler chickens at 42 days of age. On the day of hatching, 120 male and female broilers of Cobb 500 were randomly divided into three groups. The control group (1st group) of broilers received a basal diet (BD) without any feed and water additive. Both experimental groups of chicks were fed BD enriched with clove (Syzygium aromaticum L.) powder at a dose of 10 g/kg DM for 42 days. Moreover, either lemon balm (Mellisa officinalis L.) extract or agrimony (Agrimonia eupatoria L.) extract diluted with drinking water (2:1000) was given to broilers in the 2nd and 3rd group respectively. The results indicated that feeding the diets enriched with selected herbal supplements failed to affect the growth performance of broiler chickens at 42 days of age. In addition, this supplementation had no influence on the activities of SOD and GSH-Px, concentration of vitamin A and selected lipid metabolism indices. On the other hand, we observed beneficial effects on some indices of the antioxidant status (increased concentration of -SH groups and vitamin E, decreased concentration of MDA) in the blood of broilers in both experimental groups in comparison with the control group of chickens (p < 0.05). Furthermore, a slightly better antioxidant capacity was found in the blood of broilers supplied the combination of clove and lemon balm compared to clove and agrimony (vitamin E, 11.26 ± 0.73 vs. 9.73 ± 0.64 µmol/L, p < 0.05 respectively). It could be concluded that supplementation of the diet with clove flower buds powder combined with lemon balm extract or agrimony extract dissolved in drinking water has a potential to increase the antioxidant status but fails to influence either the growth performance or the selected lipid metabolism indices of broilers at the age of 42 days.

Influence of storage conditions and shotshell wounding on the hygienic condition of hunted, uneviscerated pheasant (Phasianus colchicus).

The effects of shot wounds on the hygienic conditions of pheasants (particularly those in the body cavity) were studied. Slaughtered (n = 33) and hunted pheasants (31 specimens with, and 33 specimens without shots in the body cavity) were stored uneviscerated at 0 and 4 degrees C. Specimens were taken at d 0, 3, 7, and 14. Hunted pheasants differed from slaughtered pheasants with respect to muscular hemorrhages and blood and fecal matter in the body cavity but also with regard to the presence of Escherichia coli in breast and thigh muscles. In addition, a higher thigh muscle pH (P < 0.05) was noted in hunted pheasants, with no significant (P > 0.05) increase observed during storage. Concentrations of biogenic amines in muscle tissue remained below the determination limit of 1 mg/kg for 90% of samples analyzed, with the maximum concentration for the remaining 10% of samples reaching 5.7 mg/kg, indicating a low incidence of contaminant bacteria. The observed changes in pH values and levels of biogenic amines failed to correlate with the presence or absence of shot lesions in the body cavity or abdominal region. Total aerobic counts increased significantly during storage, but the absolute numbers were consistently below 10(6) log(10) cfu/g. Although E. coli were <1 log(10) cfu/g in muscles of hunted pheasants on d 3 at 4 degrees C, counts of up to 3.7 log(10) cfu/g on d 7 at 4 degrees C indicated a loss of hygienic quality. Therefore, it is recommended that hunted, uneviscerated pheasants be stored 3 d at 4 degrees C, but not longer than 7 d after the hunt.

Comparison of various assays used for detection of beta-lactam antibiotics in poultry meat.

In this study, microbiological tests for the detection of beta-lactam antibiotics in meat and meat products were evaluated. The traditional FPT (four plate test, containing Bacillus subtilis and Kocuria rhizophila), BsDA (Bacillus stearothermophilus disc assay) and a newly developed microbiological test, Premi Test (containing Bacillus stearothermophilus) were included in the study. The limit of detection (LOD) of the Premi Test was compared with the LOD of the traditional methods. The detection limits of the tests were determined by using beta-lactam antibiotic standards dissolved in meat juice, as well as meat tissue obtained from laying hens after experimental administration of amoxicillin. Positive samples, based on inhibition of growth of the organism in the test, were confirmed by high performance liquid chromatography (HPLC). Growth inhibition in the traditional tests is visible as a clear zone on the plate, whereas for Premi Test, this is based on the absence of a colour change of the test. The LODs of antibiotics tested were as follows: Penicillin G (PENG) 5 microg kg(-1), amoxicillin (AMOX) 10 microg kg(-1), ampicillin (AMP) 25 microg kg(-1), oxacillin (OXA) 30 microg kg(-1), and cloxacillin (CLOX) 30 microg kg(-1) on the plate with Bacillus stearothermophilus. Beta-lactam antibiotics can be detected also on one plate seeded with Kocuria rhizophila, although the LODs are higher: PENG 10 microg kg(-1), AMOX 25 microg kg(-1), AMP 30 microg kg(-1), OXA 50 microg kg(-1), and CLOX 50 microg kg(-1). Premi Test was performed according to the Standard Operating Procedure intended for detection of beta-lactam antibiotics in poultry tissues with following LODs: PENG 4 microg kg(-1), AMOX 5 microg kg(-1), AMP 5 microg kg(-1), OXA 40 microg kg(-1), CLOX 50 microg kg(-1). All tests are able to detect beta-lactam antibiotics such as penicillin G, ampicillin, amoxicillin, oxacillin and cloxacillin below the maximum residue level (MRL). However, the detection limits of the Premi Test for PENG, AMOX and AMP were below the limits of BsDA and the plate containing Kocuria rhizophila.