Effects of the Mixing Process on the Rheological Properties of Waste PET-Modified Bitumen.

This paper analyses the key findings of a study devoted to PET-modified bitumen. The research program was run according to the D-optimal experimental plan based on a factorial design. Five factors, i.e., the type of polymer (source), the type of bitumen (qualitative factors), PET amount, mixing rate, and mixing temperature (quantitative factors), controlled the bitumen-polymer mixing process. The experiment included a series of determinations of bitumen's rheological characteristics obtained by MSCR (Jnr, R) and G*/sin(δ) at 50 °C, 60 °C, and 70 °C. The low-temperature properties of the composite (critical temperature) were evaluated using a BBR test. The findings showed that bitumen modification with PET primarily reduced the creep susceptibility of the bituminous-polymer mixture. The low-temperature characteristics of the modified bitumen played a secondary but essential role. The amount of polymer and the mixing rate interacted with the temperature, significantly reducing the stiffness of the composite, while the type and amount of bitumen had a substantial effect on the results obtained in the BBR test. It is worth noting that when combining bitumen and plastomer, special attention should be paid to ensuring a high level of homogeneity of the mixture by controlling the parameters of the mixing process accordingly. The tests and analyses provided crucial models (GLM), which allowed for the prediction of the plastomer-modified bitumen's low- and high-temperature properties. The resulting relationships between factors and the identification of their impact on the bitumen properties enable a better understanding of the process of bitumen modification with PET. The conclusions presented here serve as a basis for future optimisation of the modified bitumen composition. The performed studies indicate that the use of >3% plastomer in bitumen 70/100 allows for a reduction in its susceptibility (MSCR) to below 0.5 kPa-1, making it suitable for bituminous mixtures for high-traffic roads. No significant increase in critical temperature (BBR) was observed.

Relationship between semen quality and level of heterozygosity on the model of endangered population of Western capercaillie.

Sperm quality is a very important population survival factor, which can be influenced by the level of male's genetic diversity and can influence reproduction effectiveness, what may be crucial in terms of decision about local, endangered population protection plans. Thus, we analysed if heterozygosity and age influence sperm quality in the engendered European capercaillie (Tetrao urogallus) population in Polish breeding stations. Obtained results showed no relationship between normal sperm morphology and heterozygosity, as well as age and sperm morphology, but it was shown that both high and low level of heterozygosity significantly influenced the percentage of abnormal sperm. Thus, heterozygosity level can be a useful tool for population reproduction potential prediction, however, further research on how those abnormalities influence fertilization process are needed.

Effect of freeze-dried quail egg white and yolk addition to semen extender on viability of rooster sperm stored for 6 h at 4°C.

The effect of freeze-dried quail egg white and yolk addition to basic EK extender on morphology and motility of chicken broiler breeder semen was investigated. Fresh pooled semen was divided into eight parts: fresh, undiluted (control), diluted in 1:2 ratio (v/v) with basic EK extender, EK + 200 mg/ml of egg white, EK + 100 mg/ml of egg white, EK + 50 mg/ml of egg white, EK + 100 mg/ml of egg yolk, EK + 50 mg/ml of egg yolk, EK + 25 mg/ml of egg yolk. Semen samples were evaluated 15 min after dilution and after 6 h of storage at 4°C. In the fresh semen, the number of live normal sperm was the highest in semen diluted with EK + 200 mg of egg white and EK + 100 mg of egg yolk, while the highest sperm motility was in the neat semen. Semen storage reduced the number of normal sperm in all analysed semen samples. In the neat semen, the number of normal sperm decreased, in relation to the fresh not-stored samples, by 36.8% (from 72.3% to 35.5%), with EK extender by 9.2%, in samples enriched with egg white, from 8.4% (EK + 200 mg) to 10.0% (EK + 100 mg), and in EK with egg yolk addition, from 1.2% (EK + 50 mg) to 10.6% (EK + 100 mg). The highest percentage of motile sperm was observed in EK extender enriched with 50 mg of egg white (77.1%) and EK + 25 mg of egg yolk (65.3%).

Semen characteristics of ganders kept as genetic resources conservation programme.

The creation of genetic reserves of domesticated animal species and breeds almost become a necessity in the recent years, but there is a question what is the value of semen of males kept ex situ in vivo as gene conservation flocks. Presented studies assessed the response to semen collection by dorso-abdominal massage and the quantitative and qualitative semen characteristics of six goose breeds (Pomorska, Garbonosa, Kuban, Landes, Roman and Slovakia) covered by the genetic resources' protection programme. Fourteen semen collection attempts were performed per male. In each breed there were ganders with low and high sensitivity to massage. The most positive reactions were stated in Pomorska ganders (67.9%) and the least in Kuban breed (52.60%). Individual male evaluation showed that only in three breeds (Pomorska, Garbonosa and Kuban) there were individuals showing 100% susceptibility to semen collection, in some breeds only one to four positive reactions (ending with ejaculation) were noted. Results obtained indicated breed and male effect on analysed semen traits, with the exception of sperm motility. The highest number of live normal sperm (44.2% on average), sperm concentration (530 × 106 ml-1 ), the highest Semen Quality Factor (92.9) and sperm motility (50.30%) were found in semen of Kuban ganders, while the lowest values of these traits (28.7%; 230 × 106 ml-1 ; 11.4 respectively) in Slovakia ganders. The lowest sperm motility (38.3%) was observed in ganders of Roman breed, but comparing to the other breeds existing differences were not significant. Significant differences in sperm morphology between individual ganders were also observed.

Variation in Bird Eggs-Does Female Factor, Season, and Laying Order Impact the Egg Size, Pigmentation, and Eggshell Thickness of the Eggs of Capercaillie?

Despite numerous studies, intra-species variation in bird eggs is still not well explained. In the presented studies, we investigated the possible sources of this variation: female factor, laying order, and season, using the following traits of Capercaillie eggs as an example: egg size and shape, eggshell lightness, and thickness. Samples were collected for three years from three Capercaillie breeding centres located in different parts of Poland, where birds are kept in conditions close to their natural habitat and have a similar diet. The obtained results showed no significant impact of laying order on egg size, shape, pigmentation, nor eggshell thickness. This indicates that the provided nutrition ensures an adequate supply of minerals for the entire laying period. Most results did not show statistically significant differences between eggs from different breeding centres, but in one breeding centre, eggshells had lighter pigmentation. We assume the observed differences may result from females' individual features or local environmental conditions. Egg traits were highly consistent for individual females, proving that visual identification can be useful in identifying the eggs of different females.

Characteristics of semen collected from gander included in the genetic resources conservation program.

Conservative breeding ex situ in vivo is one of the most popular methods of creating genetic reserves. Unfortunately, keeping animals in small closed populations leads to inbreeding which reduces their reproductive capacity. The aim of the study was to characterize the sperm quality of 6 genetic groups of geese (northern and southern breeds) kept in Poland for many generations as genetic reserve flocks. Each breed was represented by 10 randomly selected 1-yr-old ganders, semen was collected 14 times, individually from each male, and the number of positive reactions (ended with ejaculation), semen volume, sperm concentration, and morphology were assessed. The obtained results showed a significant difference between breeds and individuals of the same group, both in males' reaction and semen quantitative and qualitative traits. From the northern breeds 193 ejaculates were obtained in total (i.e., 45.9% of all attempts), from the southern breeds 242 ejaculates (57.6%). The volume of single ejaculate varied from 0.01 mL (one drop allowing only histological smear and sperm morphology evaluation) to 0.65 mL; sperm concentration varied from 23.0 × 106mL-1 to 2376.0 × 106mL-1; the amount of total live sperm was at a similar level in all breeds (89.6%-97.7%), while live normal cells ranged between 15.2% and 67.9% depending on breed and individuals. When keeping the genetic reserves ex situ in vivo, attention should be paid to the quality of semen and males that are poor in this respect should be eliminated, in order not to lead to an excessive weakening of the reproductive capacity of the flocks covered by the genetic resources protection program.

Effects of Selected Prebiotics or Synbiotics Administered in ovo on Lymphocyte Subsets in Bursa of the Fabricius, Thymus, and Spleen in Non-Immunized and Immunized Chicken Broilers.

The effects of in ovo-delivered prebiotics and synbiotics on the lymphocyte subsets of the lymphoid organs in non-immunized 7-day-old broiler chickens and in non-immunized, sheep red blood cells (SRBC)-immunized, and dextran (DEX)-immunized 21- and 35-day-old birds were studied. The substances were injected on the 12th day of egg incubation: Prebiotic1 group (Pre1) with a solution of inulin, Prebiotic2 group (Pre2) with a solution of Bi2tos (non-digestive transgalacto-oligosaccharides), Synbiotic1 group (Syn1) with inulin and Lactococcus lactis subsp. lactis IBB SL1, and Synbiotic2 group (Syn2) with Bi2tos and Lactococcus lactis subsp. cremoris IBB SC1. In 7-day-old chicks, a decrease in T splenocytes was noticed in all groups. The most pronounced effect in 21- and 35-day-old birds was an increase in TCRγδ+ cells in Syn1 and Syn2 groups. A decrease in bursal B cells was observed in DEX-immunized Pre1 group (21-day-old birds), and in the Syn1 group in non-immunized and SRBC-immunized 35-day-old birds. An increase in double-positive lymphocytes was observed in Pre1 (35-day-old birds) and Pre2 (immunized 21-day-old birds) groups. In Pre1 and Syn1 groups (21- and 35-day-old), an increase in B splenocytes and a decrease in T splenocytes were observed. We concluded that Syn1 was the most effective in the stimulation of the chicken immune system.

Characterization and biological role of cysteine-rich venom protein belonging to CRISPs from turkey seminal plasma†.

Turkey semen contains cysteine-rich secretory proteins (CRISPs) that belong to the dominant seminal plasma proteins. We aimed to isolate and characterize CRISP from turkey seminal plasma and evaluate its possible involvement in yellow semen syndrome (YSS). YSS, which is well characterized, causes reduced fertility and hatchability. The protein was purified using hydrophobic interaction, gel filtration, and reverse phase chromatography. It then was subjected to identification by mass spectrometry, analysis of physicochemical properties, and specific antibody production. The biological function of the isolated protein was tested and included its effects on sperm motility and migration and sperm-egg interactions. Sperm motility was measured with the CASA system using Hobson Sperm Tracker. The reproductive tract of turkey toms was analyzed for gene expression; immunohistochemistry was used for protein localization in the male reproductive tract, spermatozoa, and inner perivitelline layer. The isolated protein was identified as cysteine-rich venom protein-like isoform X2 (CRVP X2; XP_010706464.1) and contained feature motifs of CRISP family proteins. Turkey CRVP X2 was present in both spermatozoa and seminal plasma. The extensive secretion of CRVP X2 by the epithelial cells of the epididymis and ductus deferens suggests its involvement in post-testicular sperm maturation. The internally localized CRVP X2 in the proximal part of the sperm tail might be responsible for stimulation of sperm motility. CRVP X2 on the sperm head might be involved in several events prior to fusion and may also participate in gamete fusion itself. Although the mechanisms by which CRVP X2 mediates fertilization are still unknown, the involvement of complementary sites cannot be excluded. The disturbance of CRVP X2 expression can serve as an etiologic factor of YSS in the turkey. This study expands the understanding of the detailed mechanism of fertilization in birds by clarifying the specific role of CRVP X2.

Hissing of geese: caller identity encoded in a non-vocal acoustic signal.

Non-vocal, or unvoiced, signals surprisingly have received very little attention until recently especially when compared to other acoustic signals. Some sounds made by terrestrial vertebrates are produced not only by the larynx but also by the syrinx. Furthermore, some birds are known to produce several types of non-syrinx sounds. Besides mechanical sounds produced by feathers, bills and/or wings, sounds can be also produced by constriction, anywhere along the pathway from the lungs to the lips or nostrils (in mammals), or to the bill (in birds), resulting in turbulent, aerodynamic sounds. These noises often emulate whispering, snorting or hissing. Even though hissing sounds have been studied in mammals and reptiles, only a few studies have analyzed hissing sounds in birds. Presently, only the hissing of small, nesting passerines as a defense against their respective predators have been studied. We studied hissing in domestic goose. This bird represents a ground nesting non-passerine bird which frequently produces hissing out of the nest in comparison to passerines producing hissing during nesting in holes e.g., parids. Compared to vocally produced alarm calls, almost nothing is known about how non-vocal hissing sounds potentially encode information about a caller's identity. Therefore, we aimed to test whether non-vocal air expirations can encode an individual's identity similar to those sounds generated by the syrinx or the larynx. We analyzed 217 hissing sounds from 22 individual geese. We calculated the Potential for Individual Coding (PIC) comparing the coefficient of variation both within and among individuals. In addition, we conducted a series of 15 a stepwise discriminant function analysis (DFA) models. All 16 acoustic variables showed a higher coefficient of variation among individuals. Twelve DFA models revealed 51.2-54.4% classification result (cross-validated output) and all 15 models showed 60.8-68.2% classification output based on conventional DFA in comparison to a 4.5% success rate when classification by chance. This indicates the stability of the DFA results even when using different combinations of variables. Our findings showed that an individual's identity could be encoded with respect to the energy distribution at the beginning of a signal and the lowest frequencies. Body weight did not influence an individual's sound expression. Recognition of hissing mates in dangerous situations could increase the probability of their surviving via a more efficient anti-predator response.

Reproductive season and male effect on quantitative and qualitative traits of individually collected Muscovy duck (Cairina moschata) semen.

Males of Muscovy duck (Cairina moschata) are mainly used for mule duck production via artificial insemination of females originated from wild mallard duck (Anas platyrhynchos); therefore, the quantity and quality of drake semen play a crucial role. The assessment results of male reaction to sexual stimulation by dummy female and basic semen characteristic (ejaculate volume, sperm concentration and morphology) of 12 individually kept Muscovy drakes carried out during the entire reproductive season are described. The male and period of the reproductive season effect on scored semen traits are documented. In total, 792 individual semen collections and evaluations were performed. The average of positive reaction in the entire reproductive season varied from 90.6% in December and April to 50.0% in July, while for individual males, it varied between 97.1% and 29.0%. Throughout the season, the ejaculate volumes ranged from 0.05 to 2.45 ml, sperm concentration from 0.15 × 109 ml-1 to 4.44 × 109 ml-1 , total number of live spermatozoa from 68.0% to 100% and live normal (properly formed, with any deformations) from 51.0% to 99.0%. Our study indicates the necessity of male breeders pre-selection before the onset of the reproductive season, and the need to leave an appropriate number of males to ensure adequate amount of semen for female insemination, especially when using Muscovy drakes (Cairina moschata) for interspecies crossing with Anas platyrhynchos ducks.

Characteristics of capercaillie (Tetrao urogallus) semen analysed with flow cytometry combined with fertility results.

In order to increase the reproductive indices of capercaillie kept in closed breeding facilities, it is necessary to constantly expand the methods of better understanding the characteristics of sperm and their fertilizing potency. The aim of the study was to analyse selected features of capercaillie sperm using flow cytometry and their connection with fertility results. The study included five males, three of which were kept in a family group with eight females and two were kept alone. For sperm viability, acrosome integrity, mitochondrial potential and DNA defragmentation were assessed. Paternity analyses were performed in order to confirm the paternity of the individual and to link the evaluated semen traits with reproductive success. Analyses carried out in the flow cytometer showed any significant differences between males in sperm characteristics. In the semen of male No. 101, the father of all chicks from the analysed family group, 91.3% of live sperm, 91.5% with intact acrosome, 83.6% with active mitochondria and 2.0% with DNA defragmentation were observed. The average fertility rate was 71.0%, and chick hatchability was 100%. Using flow cytometry in the analysis of capercaillie semen and its connection with the results of natural mating, we were able to obtain deeper knowledge about new sperm characteristics that were not examined before and which in the future may be helpful in selecting males for the reproductive flocks and developing assisted reproduction techniques.

Effect of semen extenders on viability of ISA Brown and Hubbard Flex roosters' sperm stored for 24 h.

Artificial insemination is used in almost 95% of turkey reproductive flocks and is becoming more important in chickens, particularly broiler breeders, as well as in assisted reproduction of wild birds kept in breeding centers. Diluted semen is recommended for artificial insemination. Pooled semen samples collected twice a week by dorso-abdominal massage from 2 chicken lines: laying-ISA Brown (ISA-B) and meat type-Hubbard Flex (H-F) were divided into 5 parts: neat semen and diluted in 1:2 ratio with 4 extenders: basic EK; EK + 1 µg/mL organic selenium and 8 µg/mL vitamin E; EK + 10 mg/mL of royal jelly; and EK + 0.25 g/mL of lyophilized bovine colostrum. Diluted semen samples were evaluated after 15 min and then 24 h storage at 4°C. Sperm concentration, motility, motility parameters (with Sperm Class Analyzer), and morphology were evaluated in the neat semen, whereas in diluted and stored samples, the last 3 traits were determined. In case of both lines, dilution did not affect (P > 0.05) the number of live normal cells (78.0-81.1% in ISA Brown and 73.8-68.7% in Hubbard Flex) in relation to neat semen; however, bovine colostrum addition increased (P < 0.05) the percentage of bulb head sperm (5.7 vs. 10.0% and 12.1 vs. 17.6%, for ISA and Hubbard, respectively) and decreased sperm motility (67.4 vs. 92.9% and 67.3 vs. 98.5% for ISA and Hubbard). The 24 h storage of neat semen and semen diluted with colostrum caused (P < 0.05) the unfavorable changes in all evaluated traits and both chicken lines, whereas semen dilution with remaining extenders decreased the percentage of live normal cells (by 18.8-23.4% ISA and by 20.9-25.5% Hubbard) but did not affect sperm motility (81.5-87.6% for ISA and 81.1-96.6% for Hubbard). Sperm motility and motility parameters depended both on the extender and chicken line.

Female-Male and Female-Female Social Interactions of Captive Kept Capercaillie (Tetrao Urogallus) and Its Consequences in Planning Breeding Programs.

Capercaillie behavior, both in the wild and in captivity, is poorly known due to this species' secretive way of life. Female-male and female-female social organization and interactions are especially poorly documented. The research was conducted in Capercaillie Breeding Center in Wisla Forestry District where a breeding flock is kept throughout the year. Thanks to video monitoring, we were able to observe mate choice, and then later, female-female interactions during laying and incubation period. Male individual variation in tooting latency and duration were recorded. Females' interest in males was related to males' tooting activity, but when males became too insistent and started to chase the females, the females avoided contact with them. There was a significant relationship between calendar date and when tooting starts, and between the tooting duration the female spent with a male. Two incidents of female-male aggression caused by competition for food were observed. Female intruder presence and competition for nesting place was observed in 66.67% nests. Most female-female interactions were limited to threat posturing, but fights and attempts to push out the intruder from the nest occurred as well. Such interactions may lead to nest abandonment and egg destruction, lowering the breeding success.

Impact of Prebiotics and Synbiotics Administered in ovo on the Immune Response against Experimental Antigens in Chicken Broilers.

The effect of the in ovo application of selected prebiotics and synbiotics on the humoral immune response against T-dependent (SRBC) and T-independent (dextran) antigens and delayed-type hypersensitivity (DTH) to phytohemagglutinin was studied. On the 12th day of incubation, 800 eggs (Ross 308) were divided into five groups and injected into the egg air chamber with prebiotic inulin (Pre1), Bi2tos (Pre2), a synbiotic composed of inulin and Lactococcus lactis subsp. lactis IBB SL1 (Syn1), a synbiotic composed of Bi2tos and L. lactis subsp. cremoris IBB SC1 (Syn2), and physiological saline (control group; C). The chickens were immunized twice at the 7th and 21st day of life with SRBC and dextran. A DTH test was performed on the 7th, 21st, and 35th day. The application of prebiotics and synbiotics had no significant effect on the humoral immune response. SRBC-immunized in ovo Pre1- and Pre2-treated chickens showed significantly higher serum IgG levels than the control. A significant effect on the DTH reaction was detected on the 7th (Pre1 < C) and 21st (Pre2 > Syn2) day. However; Bi2tos may transiently stimulate the cellular immune response on the 21st day. It may be concluded that the application of inulin in an egg air chamber on the 12th day of incubation may stimulate the secondary immune response. The inulin-treated group exhibited a lower mortality rate than the control group.

Supporting dataset and methods for egg sizes, eggshell thicknesses and metal concentrations measured in the shells and contents of eggs of Capercaillies Tetrao urogallus.

The dataset presented in this data paper supports "Eggshell resorption, and embryonic mobilization and accumulation of calcium and metals in eggs of wild and captive Capercaillies Tetrao urogallus" (Orlowski et al., 2019) [1]. Here we present the supplementary data on the following: (1) egg sizes, regional eggshell thicknesses (at the equator, sharp pole and blunt pole) as well as the concentrations of two major micronutrients (Ca and Mg) and eight trace elements (Cr, Cu, Mn, Fe, Co, Cd, Pb and Zn), measured in the shells and contents of eggs of captive-bred and wild Capercaillies. (2) How the proportions of elements sequestered into eggshells become depleted during embryogenesis expressed as the %change of concentrations of various elements measured in the shells and contents of eggs at different stages of embryonic advancement (early dead embryos, late dead embryos and hatched eggs). (3) The relationships between the age of dead embryos and three regional eggshell thicknesses and concentrations of different elements measured in the shells and contents of these eggs.

Selected prebiotics and synbiotics administered in ovo can modify innate immunity in chicken broilers.

BACKGROUND: A previous study showed that prebiotics and synbiotics administered in ovo into the egg air cell on the 12th day of incubation enhance the growth and development of chickens. However, the influence of this procedure on the development and efficiency of the innate immune system of broiler chickens is unclear. Therefore, the aim of this study was to evaluate whether the early (on the 12th day of embryo development) in ovo administration of selected prebiotics (inulin - Pre1 and Bi2tos - Pre2) and synbiotics (inulin + Lactococcus lactis subsp. lactis IBB SL1 - Syn1 and Bi2tos + L. lactis subsp. cremoris IBB SC1 - Syn2) influences the innate immune system. RESULTS: Chickens (broiler, Ross 308) that were treated with Pre1 exhibited a decreased H/L ratio on D7, but an increased H/L ratio was observed on D21 and D35. In the remaining experimental groups, an increase in the H/L ratio was observed on D21 and D35. The oxidative potential of leukocytes measured using the NBT test increased on D21 in Pre2 and Syn1 groups. The rate of the phagocytic ability of leukocytes increased in Pre1 and Syn1 groups on D21. The phagocytic index decreased in Pre1 and Syn2 groups on D21 and D35. Concurrently, the count of WBC in circulating blood decreased on D21 in Pre1, Pre2, and Syn1 groups. The hematocrit value was increased in Syn1 chickens on D21, in Pre1 chickens on D35, and in Syn2 chickens on both time points. CONCLUSIONS: Early in ovo treatment of chicken embryos with prebiotics and synbiotics may temporarily modulate not only the production/maturation of leukocytes but also their reactivity.

Eggshell resorption, and embryonic mobilization and accumulation of calcium and metals in eggs of wild and captive Capercaillies Tetrao urogallus.

Although it is fairly well known that eggshells are the major source of Ca and other micronutrients for developing avian embryos, potential anatomical and/or ecological correlates explaining the variation in embryo-induced depletion of micronutrients from eggshells remain to be addressed. Using unhatched eggs at various developmental stages and post-hatched eggshells of wild and captive-bred birds of a large precocial species, the Capercaillie Tetrao urogallus, we explored two major questions. (i) Do egg size, regional eggshell thickness as well as levels of two major micronutrients (Ca and Mg) and eight trace elements (Cr, Cu, Mn, Fe, Co, Cd, Pb and Zn) measured in egg shells and contents vary with embryonic age within and between wild and captive-bred bird eggs? (ii) How the proportions of eggshell elements become depleted during the entire process of embryogenesis? The eggshells of wild Capercaillies were thinner at the equator and the sharp pole, and contained more Mg and Ca, while in their contents there was more Cu, Cd and Pb compared with captive birds. All ten metal concentrations measured in the egg contents increased (from 15% to 285% more) with the age of embryos. The concentrations of six elements measured in eggshells (Cu, Mn, Fe, Co, Cd and Zn) decreased consistently (from 15 to 70% less) during embryogenesis. This suggests the presence of these elements in the innermost decalcified shell layer and their further mobilization into the embryos/egg contents. Our results strongly imply that most trace elements present in the egg contents/developing embryos are mobilized from the eggshells. We highlight the fact that knowledge of the magnitude of embryogenesis-related functional changes in the physical (such as shell thickness) and chemical traits of the eggs and eggshells of oviparous vertebrates is fundamental for drawing correct inferences regarding the response of an individual to variable environmental conditions.

Protective effects of levamisole, acetylsalicylic acid, and α-tocopherol against dioxin toxicity measured as the expression of AhR and COX-2 in a chicken embryo model.

Polychlorinated dibenzo-p-dioxins and dibenzofurans (dioxins) are classed as persistent organic pollutants and have adverse effects on multiple functions within the body. Dioxins are known carcinogens, immunotoxins, and teratogens. Dioxins are transformed in vivo, and interactions between the products and the aryl hydrocarbon receptor (AhR) lead to the formation of proinflammatory and toxic metabolites. The aim of this study was to determine whether α-tocopherol (vitamin E), acetylsalicylic acid (ASA), and levamisole can decrease the amount of damage caused by dioxins. Fertile Hubbard Flex commercial line chicken eggs were injected with solutions containing 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or containing TCDD and the test compounds. The chicken embryos and organs were analyzed after 7 and 13 days. The levels at which AhR and cyclooxygenase-2 (COX-2) proteins (which are induced during inflammation) were expressed were evaluated by performing immunohistochemical analyses on embryos treated with TCDD alone or with TCDD and the test compounds. TCDD caused developmental disorders and increased AhR and COX-2 expression in the chicken embryo tissues. Vitamin E, levamisole, ASA, and ASA plus vitamin E inhibited AhR and COX-2 expression in embryos after 7 days and decreased AhR and COX-2 expression in embryos after 13 days. ASA, levamisole, and ASA plus vitamin E weakened the immune response and prevented multiple organ changes. Vitamin E was not fully protective against developmental changes in the embryos.

Histological, histochemical and ultrastructural studies on Harderian and lacrimal glands of the Capercaillie (Tetrao urogallus major L.).

This study describes the macroscopic anatomy and the microscopic and ultrastructural features of the Harderian gland and lacrimal gland of the Capercaillies. It was conducted both on adult male and female Capercaillies. Tissue sections were stained with hematoxylin and eosin, azan trichrome, modified Mallory's trichrome, methyl green-pyronin Y, periodic acid-Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale's dialysed iron. The morphometric study of the Harderian and lacrimal glands indicated that they are both larger in male than in female Capercaillies. The histological analysis showed that the HG has a multilobar tubulo-alveolar structure with numerous lymphocytes and plasma cells. The LG has a multilobar tubulo-acinar structure without lymphocytes and plasma cells. The periodic acid-Schiff staining and alcian blue pH 2.5 staining demonstrated a mild positive reaction in the epithelial cells of the Harderian gland and weak positive reaction in the lacrimal gland. The HDI staining detected the presence of carboxylated acid mucopolysaccharides in the Harderian and lacrimal glands. Transmission electron microscopy revealed the presence of two types of secretory vesicles in the cytoplasm of both studied glands. It also showed that lipid droplets and glycogen granules were more abundant in the Harderian gland than in the lacrimal gland of this species.

Light and electron microscopic study of the eyelids, conjunctiva-associated lymphoid tissue and lacrimal gland in Bilgorajska Goose (Anser anser).

Normal structure of the accessory organs of the eye is essential for normal eye physiology. Among the most important accessory organs of the eye are the eyelids, the conjunctiva-associated lymphoid tissue (CALT) and the lacrimal gland (LG). The aim of this study was to demonstrate the histological structure of the eyelids and LG by histochemical and ultrastructural analysis. The study was performed on 13 adult female Bilgorajska geese. Eyelid samples were stained with the Alcian blue (AB pH 2.5) and periodic acid-Schiff (PAS) methods. Staining methods used for LG were AB pH 2.5, aldehyde fuchsin (AF), PAS and Hale's dialysed iron (HDI). Within the connective tissue of the eyelids, well-developed, diffuse, CALT follicles were observed, mostly under the conjunctival epithelium. Numerous lymphocytes were present within loose connective tissue. Staining of the eyelids with the PAS method demonstrated the presence of goblet cells of a mucous nature, and AB pH 2.5 staining indicated the presence of sulfated acid mucopolysaccharides. PAS staining of LG revealed the presence of secretory cells containing weakly PAS-positive granules. All epithelial cells of the corpus glandulae and the duct systems reacted positively to AB pH 2.5. HDI staining detected the presence of carboxylated acid mucopolysaccharides. Transmission electron microscopy investigations revealed two types of secretory epithelial cells in LG. Both types of LG cells contained drop-like secretory vesicles of different sizes with low or high electron density in cytoplasm, as well as small and large lipid vacuoles, and numerous small primary lysosomes.