Evaluation of the ability of colistin,amoxicillin (components of Potencil®), and fluoroquinolones to attenuate bacterial endotoxin‐ and Shiga exotoxin‐mediated cytotoxicity—In vitro studies

Escherichia coli is one of the major pathogens in humans and animals causing localized and systemic infections, which often lead to acute inflammation, watery diarrhea, and hemorrhagic colitis. Bacterial lipopolysaccharide (LPS) and Shiga exotoxins (Stx) are mostly responsible for such clinical signs. Therefore, highly effective treatment of E. coli infections should include both eradication of bacteria and neutralization of their toxins. Here, for the first time, we compared the in vitro ability of common antibiotics to decrease LPS‐ and Stx‐mediated cytotoxicity: colistin, amoxicillin (used separately or combined), enrofloxacin, and its metabolite ciprofloxacin. Three experimental scenarios were realized as follows: (a) the direct effect of antibiotics on endotoxin, (b) the effect of antibiotic treatment on LPS‐mediated cytotoxicity in an experiment mimicking “natural infection,” (c) the effect of antibiotics to decrease Stx2e‐mediated cytotoxicity. Two cell lines, A549 and Vero cells, were used to perform cytotoxic assays with the methyl tetrazolium (MTT) and lactate dehydrogenase leakage (LDH) methods, respectively. Colistin and amoxicillin, especially used in combination, were able to attenuate LPS toxic effect, which was reflected by increase in A549 cell viability. In comparison with other antibiotics, the combination of colistin and amoxicillin exhibited the highest boster or additive effect in protecting cells against LPS‐ and Stx2e‐induced toxicity. In summary, in comparison with fluoroquinolones, the combination of colistin and amoxicillin at concentrations similar to those achieved in plasma of treated animals exhibited the highest ability to attenuate LPS‐ and Stx2e‐mediated cytotoxicity.     

Interactions between fawns and does of farmed fallow deer Dama dama in the postnatal period

The aim of the present study was to describe the suckling behavior of the offspring of farmed European fallow deer Dama dama in the postnatal period. Video surveillance footage from the first 18 days postpartum was analyzed in detail to reveal that the duration of interactions between the mother and the offspring changed with the fawns’ age. The duration of mother‐offspring interactions was longest between postpartum days 12 and 18. The frequency of interactions at different times of the day varied in the analyzed periods. The results of the study could be used to increase the welfare of farmed deer.     

GLP1 and GIP are involved in the action of synbiotics in broiler chickens

Background: In order to discover new strategies to replace antibiotics in the post-antibiotic era in meat-type chicken production, two new synbiotics were tested:(Lactobacillus salivarius IBB3154 plus galactooligosaccharide(Syn1) and Lactobacillus plantarum IBB3036 plus raffinose family oligosaccharides(Syn2).Methods: The synbiotics were administered via syringe, using a special automatic system, into the egg air chamber of Cobb 500 broiler chicks on the 12 thday of egg incubation(2 mg of prebiotics + 105 cfu bacteria per egg).Hatched roosters(total 2,400) were reared on an experimental farm, kept in pens(75 animals per pen), with free access to feed and water. After 42 d animals were slaughtered. Blood serum, pancreas, duodenum and duodenum content were collected.Results: Syn2 increased trypsin activity by 2.5-fold in the pancreas and 1.5-fold in the duodenal content. In the duodenum content, Syn2 resulted in ca 30% elevation in lipase activity and 70% reduction in amylase activity.Syn1 and Syn2 strongly decreased expression of m RNA for GLP-1 and GIP in the duodenum and for GLP-1 receptors in the pancreas. Simultaneously, concentrations of the incretins significantly diminished in the blood serum(P 0.05). The decreased expression of incretins coincides with changed activity of digestive enzymes in the pancreas and in the duodenal content. The results indicate that incretins are involved in the action of Syn1 and Syn2 or that they may even be their target. No changes were observed in key hormones regulating metabolism(insulin, glucagon, corticosterone, thyroid hormones, and leptin) or in metabolic indices(glucose,NEFA, triglycerides, cholesterol). Additionally, synbiotics did not cause significant changes in the activities of alanine and aspartate aminotransferases in broiler chickens. Simultaneously, the activity of alkaline phosphatase and gamma glutamyl transferase diminished after Syn2 and Syn1, respectively.Conclusion: The selected synbiotics may be used as in ovo additives for broiler chickens, and Syn2 seems to improve their potential digestive proteolytic and lipolytic ability. Our results suggest that synbiotics can be directly or indirectly involved in incretin secretion and reception.     

Plasma and cerebrospinal fluid interleukin-1β during lipopolysaccharide-induced systemic inflammation in ewes implanted or not with slow-release melatonin

Background: Interleukin-1β(IL-1β) is important mediator of inflammatory-induced suppression of reproductive axis at the hypothalamic level. At the beginning of inflammation, the main source of cytokines in the cerebrospinal fluid(CSF) is peripheral circulation, while over time, cytokines produced in the brain are more important. Melatonin has been shown to decrease pro-inflammatory cytokines concentration in the brain. In ewes, melatonin is used to advance the onset of a breading season. Little is known about CSF concentration of IL-1β in ewes and its correlation with plasma during inflammation as well as melatonin action on the concentration of IL-1β in blood plasma and the CSF, and brain barriers permeability in early stage of lipopolysaccharide(LPS)-induced inflammation.Methods: Systemic inflammation was induced through LPS administration in melatonin-and sham-implanted ewes. Blood and CSF samples were collected before and after LPS administration and IL-1β and albumin concentration were measured. To assess the functions of brain barriers albumin quotient(QAlb) was used.Expression of IL-1β(Il1B) and its receptor type Ⅰ(Il1r1) and type Ⅱ(Il1r2) and matrix metalloproteinase(Mmp) 3 and 9 was evaluated in the choroid plexus(CP).Results: Before LPS administration, IL-1β was on the level of 62.0 ± 29.7 pg/mL and 66.4 ± 32.1 pg/mL in plasma and 26.2 ± 5.4 pg/mL and 21.3 ± 8.7 pg/mL in the CSF in sham-and melatonin-implanted group, respectively.Following LPS it increased to 159.3 ± 53.1 pg/mL and 197.8 ± 42.8 pg/mL in plasma and 129.8 ± 54.2 pg/mL and139.6 ± 51.5 pg/mL in the CSF. No correlations was found between plasma and CSF IL-1β concentration after LPS in both groups. The QAlb calculated before LPS and 6 h after was similar in all groups. Melatonin did not affected m RNA expression of Il1B, Il1r1 and Il1r2 in the CP. The m RNA expression of Mmp3 and Mmp9 was not detected.Conclusions: The lack of correlation between plasma and CSF IL-1β concentration indicates that at the beginning of inflammation the local synthesis of IL-1β in the CP is an important source of IL-1β in the CSF. Melatonin from slow-release implants does not affect IL-1β concentration in plasma and CSF in early stage of systemic inflammation.     

Engineering properties of red clover (Trifolium pratense L.) seeds

Selected engineering properties of seeds of four red clover varieties were studied. The seeds of diploid and tetraploid forms were statistically different in terms of true and bulk density, porosity, volume, 1000 seed weight, equivalent diameter and yellowness index. The volume, 1000 seed weight and equivalent diameter were found to be statistically higher for tetraploid than for diploid forms. The accuracy of varietal classification based on the physical properties of seeds was found to be most satisfactory. Varieties of Bona and Jubilatka were identified with 100% accuracy based on the physical properties of seeds. Bona was the most conducive variety for the classification of red clover seeds based on four groups of engineering properties. The relationships between groups of properties described by total redundancy did not exceed 50% for any of variable pairs.     

Magnetic resonance imaging used for the evaluation of water presence in wood plastic composite boards exposed to exterior conditions

Abstract

Two wood plastic composite (WPC) boards, one experimental and one commercial, were exposed to exterior conditions and evaluated non-destructively using a clinical magnetic resonance imaging (MRI) unit for moisture content (MC) and distribution. The experimental board was exposed in Vancouver, British Columbia, for more than 8 years, and the commercial board was exposed near Hilo, Hawaii, for 2 years. Both boards were characterized in terms of wood content, density, water uptake properties and voids content. The experimental board was additionally destructively analysed for water absorption of the WPC and MC calculated based on the wood content for verification of MRI results. MRI detected the presence of free water and its distribution in both of the WPC boards. Fibre saturation in the experimental board was found to be about 22–24%, in comparison to 25–30% present in most wood species. There was good correlation between the detection of free water by MRI and by destructive testing. Magnetic resonance images showed various major points of water entry in the WPC boards such as the support area, the cut ends, the dripping edge and the sides of the boards. For the experimental board, significant water entry also occurred at the upper exposed surface.     

Development of the neonatal B and T cell repertoire in swine: implications for comparative and veterinary immunology

Birth in all higher vertebrates is at the center of the critical window of development in which newborns transition from dependence on innate immunity to dependence on their own adaptive immunity, with passive maternal immunity bridging this transition. Therefore we have studied immunological development through fetal and early neonatal life. In swine, B cells appear earlier in fetal development than T cells. B cell development begins in the yolk sac at the 20th day of gestation (DG20), progresses to fetal liver at DG30 and after DG45 continues in bone marrow. The first wave of developing T cells is gammadelta cells expressing a monomorphic Vdelta rearrangement. Thereafter, alphabeta T cells predominate and at birth, at least 19 TRBV subgroups are expressed, 17 of which appear highly homologous with those in humans. In contrast to the T cell repertoire and unlike humans and mice, the porcine pre-immune VH (IGHV-D-J) repertoire is highly restricted, depending primarily on CDR3 for diversity. The V-KAPPA (IGKV-J) repertoire and apparently also the V-LAMBDA (IGLV-J) repertoire, are also restricted. Diversification of the pre-immune B cell repertoire of swine and the ability to respond to both T-dependent and T-independent antigen depends on colonization of the gut after birth in which colonizing bacteria stimulate with Toll-like receptor ligands, especially bacterial DNA. This may explain the link between repertoire diversification and the anatomical location of primary lymphoid tissue like the ileal Peyers patches. Improper development of adaptive immunity can be caused by infectious agents like the porcine reproductive and respiratory syndrome virus that causes immune dysregulation resulting in immunological injury and autoimmunity.     

Effects of dietary meals containing different levels of phytoestrogens on reproductive function in Bilgoraj ganders

The aim of this study was to investigate how long-term feeding of ganders with diets containing different levels of phytoestrogens affects the morphometry of testes, plasma steroid levels, sperm parameters and reproductive performance. Male Bilgoraj geese were fed diets containing grass meal (with low phytoestrogen levels) and those containing alfalfa meal and soybean meal (with higher levels of phytoestrogens). Testes were obtained from ganders at the peak of mating and the ongoing laying period as well as at the beginning of photorefractoriness. The morphometric parameters were measured in frozen testicular sections. Blood was collected monthly from December to June, and plasma testosterone, androstenedione and corticosterone concentrations were determined. During the mating season, semen was collected and its quantity and quality were investigated. In the flocks (ganders with geese) reproductive output was noted. Seasonal changes were observed in the morphology of testes and in plasma testosterone, androstenedione and corticosterone concentrations in ganders. The feeding of ganders with dietary meals containing higher levels of phytoestrogens did not affect the plasma steroid contents or the majority of the morphometric parameters of testes, except for the height of the seminiferous epithelium, which was lowered. Moreover, diets with higher levels of phytoestrogens decreased the volume of ejaculates and the percentage of normal spermatozoa. However, these changes were not reflected in the fertilisation rates.     

Evaluation of silicon as a nutritional supplement for greenhouse zinnia production

Greenhouse production of floricultural crops uses soilless substrates, which have limited amounts of plant-available silicon (Si) compared to mineral soils. The inclusion of Si supplements in greenhouse production has proven beneficial for several crops. The goal of this study was to evaluate the incorporation of Si supplements for improved flower quality of greenhouse-produced zinnia (Zinnia elegans). In the first study, quantitative horticultural traits and leaf Si, macronutrient and micronutrient concentrations were compared between Si-treated and control plants. The Si supplements evaluated were rice husk ash substrate incorporation, hydrous potassium silicate (KSiO₃) substrate incorporation or weekly KSiO₃ substrate drenches, and weekly sodium silicate (NaSiO₃) foliar sprays. Several horticultural traits differed in Si-treated plants. The flower and stem diameters were larger in plants supplemented with at least one rate of each Si supplement. Reduced plant heights and 3–5-day delays in anthesis occurred in plants supplemented with weekly KSiO₃ drenches. Leaf Si concentrations were 1.2–3.3-fold higher in Si-supplemented plants, while the macronutrients, N, S, Mg, and Ca, were lower and K was higher only in KSiO₃-supplemented plants. In a second study, the effects of weekly KSiO₃ drenches and weekly NaSiO₃ foliar sprays on the leaf resistance of zinnia plants were evaluated. Only the NaSiO₃ foliar sprays moderately increased leaf resistances. While not supporting an active role for Si in reducing transpiration, NaSiO₃ foliar sprays may form antitranspirant film that increases leaf resistance. Thus, the beneficial effects of Si supplementation on greenhouse-produced zinnia depend on the form and rate of Si used.