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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 egg white, EK + 50 mg/mL egg white, EK+ 100 mg/mL egg yolk, EK + 50 mg/mL egg yolk, EK + 25 mg/mL egg yolk. Semen samples were evaluated 15 minutes after dilution and after 6 hours 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 egg white and EK + 100 mg egg yolk, while the highest sperm motility was in the neat semen. Semen storage reduced the number of normal sperm in all analyzed 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%).

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 × 10

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.

Avian semen dilution with appropriate extender allows to prolong the fertilizing ability of sperm stored in vitro. In the present study, the impact of extenders and time of storage on morphology of Muscovy duck (Cairina moschata) drake semen were examined. Semen was collected twice a week, using male stimulation by a female method, from 12 adults (29 weeks old) drakes kept individually in cages, under controlled environmental conditions. Freshly collected, pooled ejaculates were divided into three part: neat undiluted sample, and diluted 1:1 with Schramm (SCH) or Watanabe (W) extender and stored at 4°C. Morphological examination of all samples was conducted after dilution and then, after 3 and 6 hr of storage. The storage of undiluted semen caused decrease (p ≤ .01) in live morphologically normal sperm, from 79.73% in the freshly collected ejaculates to 55.75% and to 12.12% after 3 and 6 hr of storage, respectively (average calculated for the entire reproductive season). In the semen diluted with Schramm's extender the adequate values attained 86.84, 79.65 and 61.66%, and using Watanabe extender 84.77, 83.58 and 75.25%, respectively. The period of semen storage and the type of extender caused significant (p ≤ 0,05; p ≤ 0,01) changes in sperm morphology. The longer period of storage contributed to the decrease in number of morphologically normal sperm, whereas their content in Watanabe extender after 3 and 6 hr of storage was higher (p ≤ .01) than in semen diluted in Schramm extender.

Prebiotics, probiotics, and synbiotics, delivered in ovo influence the colonization and development of the peripheral immune system in poultry. This study aimed to investigate the influence of the host genotype (broiler chickens [Ross 308] and old native Polish breed Green-legged Partridgelike [GP] chickens) on the number of B and T cells in the spleen and cecal tonsils (CT). The solution of a bioactive compound was injected in ovo on day 12 of egg incubation: prebiotics (galactooligosaccharides [GOS]), probiotics (Lactococcus lactis subsp. cremoris IBB477), and synbiotics (GOS + L. lactis). The samples were collected on day 7, day 21, and day 42 after hatching (n = 8). The number of Bu-1+ (B) cells, CD4+ cells, and CD8+ cells in the spleen and CT was estimated using immunohistochemistry. The number of germinal centers (GC) was determined in the spleen. In broilers, probiotics increased (P

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.

Embryo development and chick quality are influenced by parental genotype, age, nutrition, environment, and flock management. The aim of study was to determine if genotype, age of goose or eggs laid near the onset of egg production vs. eggs laid near the end of reproduction influence the stage of embryo at oviposition. Three experiments were undertaken. To compare genotypes (Experiment 1) 150 eggs were collected from 3-year-old commercial line White Koluda (WK) geese and from two breeds involved in a genetic resources conservation program, Zatorska (Za) and Bilgoraj (Bi). Age comparison (Experiment 2) was conducted with 200 eggs collected from 1-, 2-, 3-, and 4-year-old WK geese. To compare laying periods (Experiment 3), 150 WK eggs were collected at the first week of March and 100 at the second half of June. Eggs were stored for 72 h at 16°C, staged using Eyal-Giladi and Kochav (EGK, Roman numerals) and Hamburger and Hamilton (HH, Arabic numerals) procedures. Experiment 1: Individual breed differences were evident with Stage X EGK embryos comprising 42.4, 33.3, and 38.7% in the eggs examined from the WK, Bi, and Za, respectively. For all breeds combined, 38.8% of the embryos were in Stage X, but in the next order in WK there was stage XI (18.2%), while in geese from the genetic reserve it was stage XIII (Bi - 33.3; Za - 29.0%). Experiment 2: In eggs of 1-, 2-, and 3-year-old WK geese, the majority of embryos (38.7, 32.4 and 42.2%, respectively) were in Stage X. In contrast, in 4-year-old geese the embryos were in Stage XI (36.1%). Experiment 3: In eggs collected in March and in June most of embryos were in Stage X (33.7% and 43.6%, respectively). In addition, more developmentally advanced stages (XI-XIII) were similar in both periods. However, embryos in Stage 2 HH were only observed in eggs collected at the end of laying season. Interestingly, earlier stages (VI-IX) were observed exclusively in the eggs collected in March.

BackgroundMucosal and systemic immune responses are different strategies to cope with environmental stimuli. In ovo delivery of prebiotic, probiotic, or synbiotic into the avian embryo allows for indigenous microbiota stimulation. Intestinal microbiota in animals is responsible for immune system maturation. The genetic component is critical in host-microbiota crosstalk and immune system development. The goal of this study was to compare mucosal and systemic immune responses in two distinct chicken genotypes stimulated in ovo. ResultsThe experiment was constructed in full-factorial design and was aimed to study the effects of two chicken genotypes (chicken broilers or native chickens), four in ovo-delivered compounds (GOS/galactooligosaccharides/prebiotic, Lactococcus lactis subsp. cremoris/probiotic, or GOS+L.lactis/synbiotic, vs. physiological saline) and three different stimuli (LPS or LTA vs. physiological saline) on a panel of cytokine genes (IL-1B, IL-2, IL-4, IL-6, IL-10, IL-12p40, and IL-17) expressed in caecal tonsils and spleen. We tested significance of the main effects and their interactions. Genotype had the most significant influence on all gene expression signatures in both tissues (P < 0.001 for all genes, except P < 0.05 for IL-10 in spleen). Immune challenge was the second most significant main effect, and influenced IL-1β, IL-6 IL-10 (P < 0.001), and IL-17 genes (P < 0.05) in caecal tonsils, and all genes in spleen (P < 0.001), except IL-4 (P > 0.05). In ovo stimulation influenced IL-2, IL-4, and IL-10 in caecal tonsils (P < 0.05), as well as IL-2 and IL-12p40 in spleen (P < 0.05). ConclusionsThe mucosal and systemic immune responses of chicken broilers and native chickens showed distinct patterns. Genotype influenced gene expression signatures of all immune-related genes, but chicken broilers developed stronger immune responses than native chickens. LPS triggered both mucosal (caecal tonsils) and systemic (spleen) immune responses in chicken broilers, but only systemic (spleen) in native chickens. In ovo stimulation with bioactive compounds (especially prebiotic) modulated innate immune responses to LPS. GOS delivered in ovo induced the most pronounced responses to LPS, which validated its further application as a potent immunomodulator for in ovo applications.

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.

Background. Trunk deformity is an important manifestation of idiopathic scoliosis (IS). Patients’ perception of spinal deformity and its impact on their quality of life (QoL) are important aspects of scoliosis treatment. The aim of this study was to determine the correlation between radiographic parameters (Cobb angle), clinical parameters (angle of trunk rotation, ATR), and the type of conservative treatment used vs the perception of trunk aesthetics as well as QoL in IS patients. Material and methods. The study enrolled 90 subjects (68 females, 22 males) diagnosed with IS (age: 9-18 years, 14.3± 2.1; Cobb angle 10-66°, 22.2°±12.2; ATR 2-20°, 7.6°±3.9). The study group was divided into (1) patients treated with bracing and physiotherapy (n=35) and (2) subjects undergoing physiotherapy alone (n=55). The perception of trunk deformity was analysed with the TAPS visual scale. QoL was measured with the Polish version of the SRS-22 questionnaire. Results. The study showed that the patients’ perception of their trunk aesthetics significantly worsened with increasing Cobb angle (TAPS, rS= -0.327, p

Birds obtained by embryo engineering are used to study embryo development and to produce transgenic birds. As this method of producing birds still generate strong emotions of the public opinion head ornaments, testes and semen characteristics of sex chimera roosters were examined to check whether they differ from chickens obtained by non-manipulated methods. Measurements of head ornaments, testes and semen were correlated with each other. Semen quality factor (SQF) was calculated, as well as the level of fluctuating asymmetry (FA) of bilateral traits (wattles and testes). Positive correlation was found for comb width and wattle length and comb thickness and sperm concentration. Semen characteristics and FA did not exceed the level encounter in other chicken lines. Results obtained indicate that germline chimeras are similar in appearance of secondary sexual traits, and semen and testes characteristics to chickens produced in non-manipulated way.

The aim of the study was to describe the morphology of the upper, lower and third eyelid and characterize the organized lymphoid follicles and diffuse lymphocytes from ornamental and wild birds. The goal of these examinations was also to identify avian conjunctiva-associated lymphoid tissue (CALT) and lymphoid tissue that contained specialized high endothelial venules. The upper, lower and third eyelid from 30 species of ornamental and wild birds representing 21 families were examined under light microscopy and using scanning electron microscopy. The third eyelid in all of the examined birds was composed of a free margin, which was divided into two parts. The largest tarsal plate of the third eyelid was observed in the greater rhea (Rheimorphae), the white-tailed eagle and steppe eagle (Accipitrimorphae) and was approximately 13–15 mm wide and 9–11 mm long, respectively. In all of the examined birds, the CALT was associated with a rich network of small vessels. In addition, the presence of characteristic high endothelial venules and roundish bright endothelial cells was confirmed in the upper and lower eyelids or only in the lower eyelid (Phoenicopterimorphae, Procellariimorphae and Strigimorphae).

It is widely practiced that before mating the roosters are selected as reproducers mainly on the basis of head ornaments appearance. Relationship between males secondary sexual traits (comb and wattles) with testes size, as all of them are testosterone-dependent traits, were examined. Measurements of head ornaments and testes from two broiler breeder lines and two laying lines of chicken were correlated. The value of fluctuating asymmetry (FA) of bilateral traits was also measured. Positive correlations were found for head ornaments and testes weight in one of broiler breeder lines. No correlations were found for males of laying lines. The degree of FA did not differ between lines, except for FA of wattle length, which was higher (P≤0.05) for one of the broiler lines. Results obtained indicated that head ornaments cannot be used as the only criterion for rooster selection as reproducers.

Although it has been strongly implied that studies on structural function might resolve certain conflicting theories regarding the huge variability of coloration in avian eggs, including maculation, the fundamental physiological question about the functional role of eggshell speckling in the maternal deposition of micronutrients into maculated (pigment spots) and plain (unpigmented) eggshell regions remains largely unanswered. We measured (within the same egg) the concentrations of 2 micronutrients, the staple components of avian eggshells (calcium and magnesium), and 6 trace elements (copper, manganese, iron, cobalt, cadmium, and lead) in maculated (a superficial layer of brown pigment) and unpigmented areas of eggshells in 2 groups of Japanese Quail (Coturnix coturnix japonica) eggs representing 2 extremes of eggshell coloration: those with darker eggshells and more extensive pigment spots vs. those with bright eggshells and small, more clearly demarcated pigment spots. We found evidence that t...

The influence of a developing bird embryo on eggshell thinning is well known, however this phenomenon has been described mainly for poultry, and in wild birds is poorly understood. This study focussed on Capercaillie ( Tetrao urogallus), in which the effect of a developing embryo on the change of eggshell thickness has not been described. However, problems relating to a large number of dead embryos and eggs cracking during incubation are observed in captive breeding centres giving us the opportunity to test how the developing embryo affects the eggshell and thereby its properties, including eggshell strength and thickness. In total, 196 unhatched eggs, including 146 infertile and 50 eggs with embryos which died at various stages of development, were analysed in relation to eggshell shape, thickness and strength. Moreover, 102 posthatched eggshells were also examined. Although the strength of eggshell was affected by egg shape and shell thickness, contrary to expectations, the stage of embryonic development had no significant influence on eggshell strength. As has been demonstrated in other species, etching of eggshells by the developing embryo reduces shell thickness in Capercaillie but it has minimal effect on the strength of the eggshell. Egg viability is not compromised during the later stages of development by contact incubation with the female. The eggshell of infertile eggs or eggs containing early-dead embryos were 10% thicker than posthatched eggshells and eggs with late-dead embryos, confirming that eggshell thinning in Capercaillie is typical for precocial species.

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.

Simple Summary Probiotics, prebiotics, and synbiotics may be used as feed additives instead of banned antibiotic-based growth promoters. These bioactive compounds applied in ovo have beneficial effects on intestinal bifidobacteria, decrease the number of detrimental bacteria in the gut, stimulate the development of gut-associated lymphoid tissues (GALT), and modulate the development of lymphoid organs. The aim of our study was to determine whether the specific in ovo-delivered prebiotics and synbiotics affected the lymphocyte subsets of the bursa of the Fabricius, thymus, and spleen in non-immunized chicken broilers and in birds immunized with T-dependent (sheep red blood cells—SRBC) and T-independent (dextran—DEX) antigens. This study demonstrated that in ovo administration of prebiotics and synbiotics is a promising approach for enhancing chicken immune system functions. We conclude that a combination of inulin and Lactococcus lactis subsp. lactis IBB SL1 was the most effective of the tested compounds in the stimulation of the chicken immune system. Abstract 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.

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.

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 &lt, C) and 21st (Pre2 &gt, 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.

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.

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.

A new and efficient method of the synthesis of tributyl-β-keto- and tributyl-β-alkoxycarbonylalkylidenephosphorane via treatment of tributyl[(trimethylsilyl)- methylene]phosphorane with acid chlorides or chloroformate is described. These compounds have been studied less often than their triphenyl analogues (Maryanoff and Reitz, Chem Rev 1989, 89, 870; Taillefer and Cristau, Top Curr Chem, 2003, 229, 41; Appel, Loos, and Mayr, J Am Chem Soc 2009, 131, 704) because trialkyl-stabilized ylides are very reactive, highly perishable, and more difficult to synthesize. This paper also presents the reaction of in situ generated tributyl-β-keto- and tributyl-β-alkoxycarbonylalkylidenephosphoranes with p-nitrobenzaldehyde as a model aldehyde to obtain α,β-unsaturated ketones and esters. All reactions result in Wittig products in a completely E-stereoselective manner.

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.

The mechanism of the reaction of tributyl[(trimethylsilyl)methylene]phosphorane with benzaldehyde and its p-substituted analogues has been examined. It has been found that the electronic nature of the p-substituents in aromatic aldehydes strongly influences the stereochemical and kinetic outcome of the Peterson olefination whereas temperature substantially affects their Hammett correlation. This indicates that the Peterson olefination is a multistep reaction involving the formation of at least an oxyanion/betaine and a carbanion as intermediates. In turn, moderate Z-selectivity might be the result of “steric approach intermediate control”; however, E-selectivity seems to result from the silicon–oxygen interaction and interactions of steric substituents in competing erythro- and threo-betaines.

Contents Captive breeding has become an important tool in species conservations programmes, maintaining genetic diversity and restoring wild, endangered populations. In order to improve the reproductive efficiency of captive kept capercaillie, the purpose of the study was to determine the effect of selenium and vitamin E addition to semen extender on sperm characteristic during short-term storage. Ejaculates collected individually from four capercaillie were divided into two parts, diluted threefold with basic EK extender and EK enriched with 1 mg/ml of organic selenium and 8 mg/ml of vitamin E (EK+Se+E) and stored 24 hr at temp. +4°C. Spermatozoa morphology, motility and motility parameter were evaluated in net, diluted and stored semen samples. Significant (p

Normal tables provide an objective step-wise description of the morphological development of an embryo. Such tables have been described for the chicken, turkey, quail, and duck embryos, but there is no such staging table for goose embryos. As the goose has one of the longest incubation periods of all the poultry species and embryo mortality during incubation is relatively high, a normal table of goose embryo development would be useful in assessing the morpho-genetic status of the goose embryo before and during incubation. In this study, embryos were isolated from commercial White Koluda goose eggs stored no longer than four days in a cool room (18°C) prior to incubation and after 4, 8, 12, and 16 h of incubation. Embryo staging was based on the normal tables described for the chicken by Eyal-Giladi and Kochav (EGK) and Hamburger and Hamilton (HH). Goose embryos from unincubated eggs were at Stage X and XI EGK and after 16 h of incubation the majority of embryos were between Stages 2 and 4 HH. Our results suggest that while the stage of development of the embryo in the unincubated goose egg is similar to that reported for the chicken, although the diameter of goose embryo is slighter larger. Following incubation, a goose embryo advances more slowly than a chicken embryo up to 16 h of incubation.

Dioxins have adverse and multifaceted effect on body functions. They are known to be carcinogens, immunotoxins, and teratogenic agents. In vivo, transformation of dioxins occurs after their interaction with the aryl hydrocarbon receptor (AhR) and leads to formation of proinflammatory and toxic metabolites. The aim of this study was to verify whether α-tocopherol (vitamin E) and acetylsalicylic acid (ASA), could reduce the damage caused by the action of dioxins. Fertile chicken eggs were injected with a solution of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), followed by the injection of α-tocopherol or acetylsalicylic acid. Organs such as heart and liver were dissected from the chick embryos at d 13 and 19 of development and subjected to immunohistochemical analysis of presence of advanced glycation end products (AGEs) and nuclear factor kappa B (NFκB) in tissues. The AGEs were used as the marker for exposure to dioxins, since it is well established that their level increases in dioxin-damaged tissues. Formation of AGEs was evaluated in embryos exposed to dioxin and treated with vitamin E and/or ASA (against dioxin-exposed, untreated controls). We have found that TCDD causes developmental disorders and increases the level of AGEs in chick embryo tissues. The use of such pharmacological agents as vitamin E, ASA, and combination of ASA and vitamin E, inhibited formation of the AGEs in 13-day-old embryos and reduced the AGEs level in embryos after 19 d of the development.

The aim of the present study was to demonstrate the histological and histochemical structure of the Harderian gland in wild and hybrid of wild and domestic birds. The samples were stained with haematoxylin-eosin, methyl green-pyronin Y, periodic acid-Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale's dialyzed iron staining's. In both species, the glands had multilobar tubuloacinar structure type I. The Harderian gland was located in the orbit near the inter-orbital septum, between the medial rectus muscle, the pyramidal muscle of the third eyelid, and the ventral oblique muscle. In the common pheasant, the gland was wider in the proximal and distal part. The common pheasant had more elongated lobes of the Harderian gland than in the hybrid. In the common pheasant, the glandular cells presented darkly-stained serous secretion and lightly-stained mucous secretion. In the hybrid, the glandular cells presented seromucous secretion. The central lobar space, interacinar space, and apical parts of the acini of the Harderian glands were filled with many lymphocytes and plasma cells, particularly in the common pheasant, where centers of all large lobes were abundantly filled with plasma cells. The plasma cells dominated in common pheasant's Harderian gland, while in the hybrid, lymphocytes and plasma cells were present at similar quantities. The cells positive for periodic acid of Schiff staining were dominant in hybrid. Periodic acid-Schiff, Hale's dialyzed iron and alcian blue pH 2.5 stainings demonstrated acid-carboxylated mucopolysaccharides in the glandular cells cytoplasm of the examined birds.

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.

The aim of the present study was to determine the influence of chicken semen cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activities. Pooled semen from 10 Black Minorca roosters was used in the study. Semen samples were subjected to cryopreservation using the "pellet" method and dimethylacetamide (DMA) as a cryoprotectant. In the fresh and the frozen-thawed semen sperm membrane integrity (SYBR-14/propidium iodide (PI)), acrosomal damage (PNA-Alexa Fluor(®)488) and mitochondrial activity (Rhodamine 123) were assessed using flow cytometry. Malondialdehyde (MDA) concentration, catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in sperm cells and seminal plasma by spectrophotometry. All sperm characteristics evaluated using flow cytometry were affected by cryopreservation. After freezing-thawing, there was significant (P < 0.01) reduction in sperm membrane integrity, sperm acrosome integrity and mitochondrial activity. Following cryopreservation, MDA concentration significantly increased in chicken seminal plasma and spermatozoa (P < 0.01, P < 0.05). The CAT activity in seminal plasma significantly decreased (P < 0.05), while intracellular activity of this enzyme did not significantly change in frozen-thawed semen. In seminal plasma of frozen-thawed semen the significant increase (P < 0.01) in GPx activity was detected. Whereas GPx activity in spermatozoa remained statistically unchanged after thawing. The SOD activity significantly increased (P < 0.01) in cryopreserved seminal plasma with simultaneous decrease (P < 0.01) of its activity in cells. In conclusion, this is probably the first report describing the level of antioxidant enzymes in frozen-thawed avian semen. The present study showed that the activity of CAT, GPx and SOD in chicken semen was affected by cryopreservation, what increased the intensity of lipid peroxidation (LPO). Catalase appeared to play an important role in the sperm antioxidant defense strategy at cryopreservation since, opposite to SOD and GPx, its content was clearly reduced by the cryopreservation process. Change in the antioxidant defense status of the chicken spermatozoa and surrounding seminal plasma might affect the semen quality and sperm fertilizing ability.

Mazowiecka 28, 85-084 Bydgoszcz, Poland(Received 4 July 2011; revised version 27 October 2011; accepted 12 March 2012) ABSTRACTThe effect of the level of maize distiller’s dried grains with solubles (DDGS) in diets for ducks and duration of the fattening period on feed efficiency, body weight, carcass and meat quality was studied. One hundred and sixty commercial Pekin ducklings were divided randomly into four groups (four replicates, each). From 1 to 21 days of age all birds received the same commercial feed, then from day 22 to 56 the ducks were fed with a diet containing one of four levels of DDGS (%): 0 (control), 15, 25 or 30. All ducks were weighed individually at 1, 21, 49 and 56 days of age and feed intake and feed efficiency were calculated. At 49 and 56 days of rearing, 10 birds with body weights close to the average weight for the group were chosen from each group, slaughtered and the following parameters were evaluated: weight of eviscerated carcass with neck, neck without skin, wings with skin, breast and leg muscle, edible giblets (heart, liver, gizzard), skin with subcutaneous fat, abdominal fat, remainder of carcass, as well as the physical and chemical characteristics of breast meat. Inclusion of maize DDGS in amounts up to 25% in the diet of ducks from day 22 of rearing had no adverse effect on the variables studied, regardless of age at slaughter. Moreover, it allowed decreasing the soyabean meal and wheat contents from 18.0 and 41.6% in the control feed to 10.0 and 24.1% in the 25% DDGS feed, respectively.KEY WORDS: DDGS, Pekin ducks, body weight, feed efficiency, carcass quality, breast meat

Experiments on semen collection and preservation were undertaken by Wroclaw University of Environmental and Life Sciences and Forestry Wisla, Poland to assist in the protection of the capercaillie (Tetrao urogallus L.) and to create an ex situ in vitro cryobank. Semen was collected from 11 captive-bred males, using dorsoabdominal massage. Ejaculates once obtained were diluted 3-fold at room temperature with EK diluent and then a number of them were stored at 4 °C for 18, 24, and 48 hours, while the remaining ejaculates were equilibrated with 6% dimethylacetamide and frozen by pipetting, drop-by-drop directly onto a liquid nitrogen surface. Frozen pellets were thawed at 60 °C in a water bath after 4 to 28 mo of storage. In total, 103 individually collected ejaculates (54 stored as liquid and 49 frozen in liquid nitrogen) were of appropriate value for further processing. The volume of ejaculates varied from 30 to 240 μL; spermatozoa concentration from 70 × 10(6) mL(-1) to 1950 × 10(6) mL(-1). The total amount of live spermatozoa in the fresh semen varied from 85.3% to 99.0%, of which from 41.1% to 85.3% were morphologically normal. Among morphologically abnormal forms, bulb-head (5.6% to 36.0%) and midpiece deformations (1.3% to 16.6%) were the most frequent. Dilution and semen storage up to 24 h at 4 °C did not affect the semen quality, as far as motility and sperm morphology are concerned. A significant (P < 0.05) decrease in total live (94.9 vs. 91.7%) and live normal cells (66.4 vs. 56.7%) was observed after 48 h. About 30% to 40% of spermatozoa remained motile. Cryopreservation significantly decreased (P < 0.05) the total number of live and live normal spermatozoa however, in relation to the fresh semen, their average content was 44.1% and 37.4%, respectively. Significant (P < 0.05) individual differences were observed in the quality of the fresh, liquid stored and the frozen-thawed semen assessed in terms of spermatozoa motility and morphology. After a single insemination with thawed semen containing 9.7 million live normal cells, 80% fertility and 100% hatchability were achieved. The obtained results indicate for the first time that there is the potential to use liquid stored and cryopreserved capercaillie semen to support conservation measures for the maintenance of genetic diversity, as well as to increase the number of reintroduced progeny of this endangered grouse species.

The objective of the present experiments was to produce the intergeneric hybrids of domesticated and wild goose via artificial insemination with fresh and frozen-thawed semen. The experiments were carried out during two successive goose reproductive seasons, on eight five-year-old Canada Goose (Branta canadensis L.) males used as semen donors and 16 two-year-old White Koluda geese designated to fertility tests. Pooled semen was collected twice a week by the dorso-abdominal massage. In freshly collected semen, ejaculate volume, color, consistency, degree of fecal or blood contamination, spermatozoa concentration, motility, and morphology were evaluated. Part of the semen collected in the first year of the experiment (Experiment 1) was used for geese insemination with fresh semen, while the remainder was frozen. In Experiment 2 all samples were subjected exclusively to freezing procedure. Geese were inseminated once a week with fresh semen in a dose of 80 μl or 160 μl, and twice a week with frozen-thawed semen in a dose of 80 μl (160 μl per wk) or 100 μl (200 μl per wk). Eggs were set weekly and incubated up to hatching. The volume of ejaculates varied from 0.100 to 0.470 ml; spermatozoa concentration from 140 to 310 million ml−1; progressive movement was observed in 40 to 60% of spermatozoa; the percentage of total live spermatozoa ranged from 69.3 to 92.0%, the highest percentage (34.0–68.3) was represented by live normal spermatozoa and those with bulb-head (13.3–41.0). Cryopreservation caused a decrease in percentage of motile cells to 30%; total live spermatozoa contribution by 27.2%p, including those live normal by 15.9%p (in relation to the fresh semen), bulb-head spermatozoa by 10.9%p, and increase (by 5.9%p) in number of spermatozoa with other deformations. Goose insemination 1×/week with fresh semen containing about 10.3 million live normal spermatozoa resulted in 66.7% of fertile eggs and with dose higher by 2.8 million spermatozoa (on average) the fertility increased by 20.9%p (up to 87.6% on average). Hatchability from set and fertile eggs was 55.9% and 83.9% vs. 66.3% and 75.6%, respectively. After twice a week insemination with frozen-thawed semen containing about 10.2 million live normal cells 58.2% eggs were fertile; hatchability from set eggs was 42.8% and from fertile eggs 71.7%, while insemination dose increase by 2.7 million spermatozoa per week caused a fertilization increase by 3.8%p (62.0% on average), this increase was not statistically significant, but hatchability from the fertile eggs (95.4%), was significantly (P < 0.05) higher. The use of AI with fresh semen in the creation of intergeneric hybrids of Canada goose males and White Koluda females allows a high level of egg fertility to be obtained. Furthermore, one limitation which is the short reproductive season of the Canada goose may be overcome by the use of cryopreserved semen.

Efficient synthesis of alkenylphosphonium salts via treatment of enolizable aliphatic aldehydes with tributyl[(trimethylsilyl)methylene]phosphorane has been described. The simple way for preparation of tributyl[(trimethylsilyl)methylene]phosphorane is also presented.

In Poland Capercaillie (Tetrao urogallus L.) is one of the most seriously endangered grouse species. The ability of semen collection and its utilization for Capercaillie female insemination would allow overcoming some fertility problems observed in captive-bred populations and thus reduce the rate of loss of genetic diversity. The present experiment was carried out on 13 individuals: eight males were kept with females and five alone. From each male, semen was collected four times, every second day, and overall semen appearance (color, viscosity), ejaculate volume, spermatozoa concentration, motility and morphology were examined. Ejaculates suitable for artificial insemination (AI) were obtained from 11 individuals. The volume of ejaculates varied from one drop (noted as 0.010 ml) to 0.180 ml, whereas spermatozoa concentration varied from 100 × 106 ml−1 to 1950 × 106 ml−1. The total amount of live spermatozoa for males kept with females varied from 82.0 to 98.3% (92.9% on average) and among them, from 38.7 to 82.0% were morphologically normal (67.6% on average), whereas for solitary males these values were the following: from 93.7 to 98.7 of total live (96.3% on average) and from 45.0 to 85.3% live normal cells (65.7% on average). No significant group effect was observed for above traits. Semen from males kept with females contained significantly (P

The mefenamic acid sodium, potassium, and calcium salts with general formulae [Na(mef)(H 2 O) 2 ] n · n H 2 O, [K(mef)(H 2 O)] n and [Ca(mef) 2 (H 2 O) 2 ] n · n H 2 O have been synthesised, studied by X-ray crystallography, 1 H and 13 C NMR and IR spectroscopy. The complex salts are air stable and soluble in water. During heating the Na and K complexes melt in the complexed water and next recrystallise in anhydrous form. In the solid state all salts create one-dimensional coordination polymers. The central atoms are five, six and seven coordinated, respectively, for Na, K and Ca complexes. In all structures exist O H⋯O, N H⋯O and C H⋯O hydrogen bonds. The vibrational analysis has been carried out for mefenamic acid and its three coordination polymer compounds on the basis of experimental results as well as quantum mechanical calculations. The theoretical and experimental vibrational frequencies are similar and reveal characteristic vibrations for all IR active oscillators. In the IR spectra of salts exist strong bands at ca. 1365 and 1600 cm −1 typical for carboxylate groups.