Genotype-dependent development of cellular and humoral immunity in the spleen and cecal tonsils of chickens stimulated in ovo with bioactive compounds.

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 < 0.05) the number of CD4+ cells in the CT on day 7. On day 21, prebiotics raised (P < 0.01) the number of cells involved in cellular immunity in the CT (CD4+ and CD8+ cells) and spleen (CD8+ cells). On day 42, it was synbiotics that stimulated the colonization of both the CT and spleen by B cells, but colonization of the spleen only by CD4+ and CD8+ cells. In GP chickens, synbiotics enforced the cellular immunity (CD4+ or CD8+ cells) in the spleen at all time points. Synbiotics also stimulated the GC appearance on day 21 and day 42. In GP chickens, the influence of bioactive compounds on colonization of the CT was very limited. In broilers, we determined pronounced and age-dependent effects of prebiotics and synbiotics on the number of B and T cells in both the CT and spleen. In GP chickens, the most potent compound was synbiotics, which stimulated cellular immunity in the spleen but not in the CT. However, given the long-term effects on adaptive immune cells, synbiotics were the most potent compounds in both chicken genotypes.

Stage of goose embryo development at oviposition depending on genotype, flock age, and period of laying.

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.

Secondary sexual traits and semen characteristic of chicken germline chimeras.

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.

Effect of selenium and vitamin E addition to the extender on liquid stored capercaillie (Tetrao urogallus) semen quality.

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 < .05) differences between individual males were stated in relation to the majority of traits evaluated in the freshly collected semen. Comparing to the fresh semen, a significant (p < .05) decrease in percentage of live sperm in total (by 3.8% points on average) has been observed in samples diluted by EK extender, while in semen diluted with EK+Se+E extender this decrease was lower (1.5%pts on average) and not significant. Also per cent of motile sperm in EK+Se+E extender was higher (p < .05) then in EK (71.6% vs. 58.9%), but taking into account the values of individual males, both extender and male effect on liquid semen storage become apparent. Obtained data allow concluding that selenium and vitamin E addition to EK extender had positive effect on morphology and motility of capercaillie semen stored 24 hr at 4°C and can be recommended for similar studies carried out on other Galliformes species.

Expression of advanced glycation end-products and NFκB in chick embryos exposed to dioxins and treated with acetylsalicylic acid and α-tocopherol.

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.

Goose embryonic development from oviposition through 16 hours of incubation.

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.

The Usefulness of Captive Kept Capercaillie (Tetrao urogallus L.) as the Semen Donors for Artificial Insemination and Gene Pool Preservation In vitro.

Captive breeding of birds threatened by extinction in zoological gardens or other closed aviary centres is one of the methods allowing their protection and gene pool preservation ex situ in vivo. Such birds are usually kept in captivity lifelong and serve as parents of several new generations that can be further released into natural environment, or males are used as semen donors for artificial insemination and gene banking. Therefore, the fecundity of such flocks (number of laid egg and spermatozoa quantity and quality) is very important. The aim of this study was to evaluate the usefulness of captive kept capercaillie (Tetrao urogallus L.) as semen donors in three subsequent reproductive seasons, based on the assessment of manually collected semen quality. Male response to dorso-abdominal massage, ejaculate volume, sperm concentration, motility and morphology were evaluated individually at three succeeding years. Depending on individual male properties and year of collection, the number of positive reactions to semen collection attempts (i.e. ending with ejaculation) varied from 44.4% to 100.0%; single ejaculate volume ranged from 10 to 300 µl, spermatozoa concentration from 10 × 10(6) per ml to 3520 × 10(6) per ml and percentage of live morphologically normal spermatozoa from 19.3 to 80.3%. The highest average value (66.7) of semen quality factor (SQF) was noted for a 2-year-old male (varying from 1.9 to 258.1), while the lowest for ten- (4.8; varying from 0.1 to 17.0) and 7-year-old (6.6; varying between 0.6 and 13.6). Assuming that for AI purposes, the ejaculate quality has to be at minimum 10 SQF, obtained results indicate that majority of capercaillie kept in captivity, both young (2-3 years old) and older (up to 10 years old), can be valuable semen producers in succeeding seasons.

The ostrich (Struthio camelus) ejaculate--effects of the method of collection, male age, month of the season, and daily frequency.

1. Over three breeding seasons on a farm in Poland, semen was collected from 11 ostriches using the dummy and the teaser method to study the effects of the method of collection, male age, month in the breeding season, and daily collection frequency on ejaculate characteristics. 2. A total of 259 ejaculates were collected, with an average volume of 1·28 ± 0·6 (±SEM) ml. Sperm concentration was 3·34 ± 0·08 × 10(9)/ml, the total number of spermatozoa 4·32 ± 0·22 × 10(9), and motility 4·56 ± 0·04. 3. There was no difference in ejaculates collected by the dummy and teaser methods, but the between-individual variation was considerable. Ejaculate characteristics increased with male age and varied between months, with little evidence for seasonal decline. Daily collections for 10 days did not affect sperm output. 4. The results open up avenues for further research on development of a viable protocol for artificial insemination in ostriches and efficient semen storage. 5. The between-male variation suggests that the ejaculate output could be maximized through selection.

Effects of sex and inclusion of dried distillers grains with solubles on slaughter yield and meat characteristics of Pekin ducks.

1. The effects of dried distillers grains with solubles (DDGS) dietary inclusion concentration, and sex, on body weight, slaughter efficiency and meat characteristics of Pekin ducks (Anas platyrhynchos f. domestica, strain P55) were studied. 2. Sexed ducklings (n = 160) were divided randomly into 4 groups (each with 4 replicates). From d 1 to d 21, all the birds received the same commercial feed, then from 22 to 49 d of age the ducks were fed in the following groups: control (commercial feed) and three experimental groups (15%, 25% and 30% inclusion of DDGS). All ducks were weighed individually at d 1, 21 and 49. On the day of slaughter, 5 males and 5 females, of body weight close to the average weight for group and sex, were selected from each group, slaughtered and the following parameters were evaluated: slaughter yield, weight, and percentage of particular elements of carcase, physical and chemical characteristics of meat. 3. The results obtained showed that DDGS included from 22 to 49 d of rearing, at concentrations up to 30%, in a commercial Pekin duck diet did not affect the live body weight, slaughter yield, weight, and percentage of breast and leg muscle, skin with subcutaneous fat, and abdominal fat. There were no differences in physical characteristics (pH(15), pH(24,) meat colour values L*, a*and b*, and hygroscopicity) of breast muscle, as well as in cholesterol content. DDGS addition at 30% significantly increased fat content in male, and crude protein in female, breast meat. Sex effect was observed only in a few traits and was diet dependent. Final body weight of females fed 30% DDGS was significantly lower than males; in the control and 15% DDGS group females had higher percentage of skin with subcutaneous fat. 4. The results obtained, and relative costs of feeds produced, allows the recommendation of DDGS addition at up to 30% to commercial Pekin duck diets from 22 d of age.

Efficacy of evaluation of rooster sperm morphology using different staining methods.

This work focused on inexpensive methods of evaluation fowl sperm morphology, based on eosin-nigrosin smears, which can determine disorders in spermatogenesis and can be recommended for evaluating the fertilising potency and selecting males in flocks reproduced by artificial insemination. Four fowl breeds (Black Minorca, Italian Partridge, Forwerk and Greenleg Partridge) were used to determine the efficacy of sperm morphology evaluation using four eosin-nigrosin staining methods (according to Blom, Bakst and Cecil, Morisson, Jaskowski) and three examiners of different experience (high, medium, novice). There were significant (P< or = 0.01) differences in sperm morphology between Blom's staining method and those of Bakst and Cecil, Morisson or Jaskowski, irrespective of fowl breed and examiners experience. Blom stain caused sperm head swelling and showed a drastic reduction in the proportion of live spermatozoa with normal morphology. The staining method had a greater influence on sperm morphology evaluation than the experience of the examiners.

Correlations between body measurements and tissue composition of oat-fattened White Koluda geese at 17 weeks of age.

1. The aim of the present study was to evaluate the yield and tissue composition of carcases from White Koluda ganders (males) and geese (females), and to determine the relationships between body measurements and carcase tissue composition. 2. The experiment was carried out on 200 geese (100 males and 100 females) reared to 14 weeks of age and then fed with oats for the next 3 weeks. Live body weight and body dimensions, slaughter yield, weight and percentage of muscles and skin with fat in carcase were measured and correlation coefficients were calculated between body measurements and slaughter values and carcase tissue composition. 3. White Koluda geese reared to 17 weeks of age were characterised by high body weight (6705 g) and good muscle yield (29.9%). Males had greater body weight, musculature and fatness than females. Sternum length and breast circumference, width and depth were good indicators of carcase muscle weight in 17-week-old geese. 4. Negative coefficients of correlation between sternum length and weight of skin with subcutaneous fat indicate that increased selection pressure in pedigree flocks of geese on sternum length should be paralleled by reduced carcase fatness in these birds.

Semen collection in the ostrich (Struthio camelus) using a dummy or a teaser female.

1. We investigated the feasibility of training male ostriches to ejaculate into an artificial cloaca (AC) using a teaser female or a dummy. 2. Male ostriches develop desirable behaviour patterns that allow them to be trained to the teaser and the dummy female. Training success can be high as both methods rely on natural stimulation and voluntary ejaculation into the AC provided suitable individuals are identified. 3. The key factors to successful training appear to be: the teaser method - temperament of male ostriches, the crouching behaviour of the female ostriches for the semen collector and acceptance of the collector by the male; dummy method - courtship of humans, learning ability of ostriches to habituate and then to mount a dummy. 4. Both methods are reliable, yield ejaculates of high quality and give reproducible results. The ejaculate volume was 1.09 +/- 0.13 ml, the concentration of spermatozoa 4.21 +/- 0.27 x 10(9)/ml, the total number of spermatozoa 4.67 +/- 0.62 x 10(9) and motility 4.3 +/- 0.1. 5. Commercial ostrich production that relies on natural mating can take advantage of those methods to develop artificial insemination technologies.

Comprehensive two-dimensional GC (GCxGC) qMS analysis of tetrachloronaphthalenes in Halowax formulations.

A combination of non-polar and shape selective columns in a comprehensive two-dimensional GC (GCxGC) system consisting of DB-5MS and LC-50 as the 1st and 2nd dimension columns was used to separate 22 tetrachloronaphthalene isomers. These columns enabled separation of all possible isomers of tetraCN found in the technical chloronaphthalene Halowax formulations into 18 peaks covering 15 single separated isomers and 7 co-eluting in triplicate (1,2,4,6-/1,2,4,7-/1,2,5,7-tetraCN; nos. 33/34/37) and in pairs (1,2,3,7-/1,2,4,5-tetraCN; nos. 30/32 as well as 1,3,5,8-/1,3,6,8-tetraCN; nos. 43/45). Twelve isomers of tetraCN resolved as single compounds by the GC x GC system used were found in Halowax 1001, 1013, 1014 and 1099 as well as in Equi-Halowax mixture. Three other tetraCNs that could be also resolved as single compounds were not detected in these materials, while the remaining 7 tetraCNs, which co-eluted, waits a further separation. The majority of constituents of the tetraCN homologue group of the Halowax formulations are isomers such as 1,4,5,8- (no. 46), 1,2,5,8- (no. 38), 1,2,4,8-tetraCN (no. 35), co-eluting 1,3,5,8-tetraCN (no. 43 in a pair nos. 43/45) and also co-eluting 1,2,4,6-/1,2,4,7-/1,2,5,7-tetraCN (nos. 33/34/37). The less abundant by amongst of tetraCNs in these mixtures are 1,4,6,7- (no. 47) and 1,3,5,7-tetraCN (no. 42). The remaining seven isomers, i.e., 1,2,3,4- (no. 27), 1,2,3,5- (no. 28), 1,2,3,6- (no. 29), 1,2,5,6- (no. 36), 1,2,6,7- (no. 39), 1,2,6,8- (no. 40) and 1,2,7,8-tetraCN (no. 41) are usually minor by quantity, while 1,2,3,7-/1,2,4,5-tetraCN (nos. 30/32) are also minor, or one of them is absent. Three tetraCNs not found in the Halowax formulations are isomers such as 1,2,3,8-tetraCN (no. 31), 1,3,6,7-tetraCN (no. 44) and 2,3,6,7-tetraCN (no. 48).

HRGC/HRMS analysis of chloronaphthalenes in several batches of Halowax 1000, 1001, 1013, 1014 and 1099.

Chloronaphthalene (CN) congeners and homologue groups have been quantified in up to three batches of several types of technical CN formulations of the Halowax series (Halowax 1031, 1000, 1001, 1013, 1014 and 1099), to elucidate possible batch-to-batch compositional variations. Using isotope dilution and HRGC/HRMS a relatively large variation in CN congeners and homologues composition among the batches of particular types of the Halowax formulations could be noted, and also when compared to the composition declared by the manufacturer. Depending on the type of the Halowax formulation and its batch in total up to 54 peaks from chloronaphthalenes (Agilent Ultra 2 liquid phase), which represented 70 of 75 CN congeners theoretically possible, could be found in these mixtures. These congeners represented all CN homologue groups from mono- to octaCN but some co-eluted. A co-eluting congeners were such as: 1,4-/1,6- (nos. 5/7), 1,5-/2,7- (nos. 6/12), 2,6-1,7- (nos. 11/8) of diCNs; 1,3,6-/1,3,5- (nos. 20/19), 1,3,7-/1,4,6- (nos. 23/24), 1,6,7-/2,3,6- (nos. 25/26) of triCNs; 1,2,5,7-/1,2,4,6-/1,2,4,7- (nos. 37/33/34), 1,3,6,8-/1,2,5,6- (nos. 45/36), 1,2,3,5-/1,3,5,8- (nos. 28/43), 1,2,3,4-/1,2,3,7- (nos. 27/30), 1,2,5,8-/1,2,6,8- (nos. 38/40) of tetraCNs; 1,2,3,5,7-/1,2,4,6,7- (nos. 52/60), 1,2,3,5,8-/1,2,3,6,8- (nos. 53/55) of pentaCNs; 1,2,3,4,6,7-/1,2,3,5,6,7- (nos. 66/67), 1,2,3,4,5,7-/1,2,3,5,6,8- (64/68) and 1,2,4,5,6,8-/1,2,4,5,7,8- (nos. 71/72) of hexaCNs. Absent in the Halowaxes were CN congeners such as 1,3,8-triCN (no. 22) (<0.0002 mg/g), 1,3,6,7-tetraCN (no. 44), 1,2,3,6-TetraCN (no. 29), 1,2,3,6,7-pentaCN (no. 54) and 1,2,3,6,7,8-hexaCN (no. 70) (<0.0005 mg/g).

Chloronaphthalenes composition of several batches of Halowax 1051.

Halowax 1051 is the highest chlorinated technical chloronaphthalene mixture among seven known formulations of the Halowax series. Octa- and heptaCN homologue groups are the main CN constituents of Halowax 1051 with declared 90% and 10% contents, respectively. In this study, using an isotope dilution technique and HRGC/HRMS, octaCN and heptaCNs contents of six batches of Halowax 1051 were between 82-93% and 6.2-17%, respectively. Also mono- to hexaCNs were found in Halowax 1051, and their content more or less varied according to the batch; also, the abundance of a particular CN congeners varied. Tetra-, penta- and hexaCNs have been found in all six batches of Halowax 1051 examined, and their contents varied between 0.0024-0.77%, 0.031-0.22%, and 0.21-0.82%, respectively. TriCNs have been found in three of six batches, and mono- and diCNs in two of six batches with 0.0020-0.40, 0.0017-0.25 and 0.0012-0.34% for positive findings, respectively. 2,3-DiCN (no. 10), 1,8-diCN (no. 9) at < 0.0002 mg/g, 1,6,7-/2,3,6-triCNs (nos. 25/26), 1,3,8-triCN (no. 22) at < 0.0002 mg/g, 1,3,6,7-tetra (no. 44), 1,2,3,6-tetra- (no. 29), 1,2,7,8-tetraCN (no. 41) and 1,2,3,6,7,9-hexaCN (no. 70) at < 0.0005 mg/g have not been found in Halowax 1051.

Effect of collection frequency on quantitative and qualitative characteristics of pigeon (Columba livia) semen.

The aim was to estimate the optimal frequency of semen collection from pigeons in relation to ejaculate volume, sperm concentration, total spermatozoa in ejaculate and percentage of live morphologically normal cells. The study was carried out on 455 ejaculates collected from two groups of pigeons, each of 10 males (group I: meat-type breed; group II: fancy pigeon). The birds were selected and kept individually in cages under a natural photoperiod. A two-person technique was used for semen collection (lumbo-sacral and cloacal region massage). Semen was collected once, twice or three times per week. Colour, consistency and volume of ejaculates were evaluated macroscopically immediately after collection. Sperm concentration and total number of cells in the ejaculate were estimated after dilution with Ringer's solution. A live-dead stain technique (nigrosin-eosin) was used to determine the percentage of live and normal spermatozoa. Semen collected 3x/week was of high quality. The average volume of a single ejaculate was small (21 microl in group I and 19 microl in group II), but sperm concentration was high--1.58 x 10(9)/ml and 1.96 x 10(9)/ml, respectively. The mean number of spermatozoa per ejaculate was 30.48 x 10(6) in group I and 39.49 x 10(6) in group II. An increased percentage of live and normal spermatozoa in semen collected more frequently was also observed. Collecting pigeon semen 3x/week provides spermatozoa in larger amounts and of better quality than less frequent collections (1x/week or 2x/week) and is recommended for obtaining more insemination doses.

By-side impurities in chloronaphthalene mixtures of the Halowax series: all 12 chlorobenzenes.

Total chlorobenzene content of the Halowax mixtures ranged from 1100 to 9800 ng/g. A lower chlorinated Halowax 1031 and 1000 were a several times more contaminated with chlorobenzenes than higher chlorinated Halowax 1001, 1099, 1013, 1014, or 1051. Depending on the type of the Halowax mixture the most contributing amongst of chlorobenzene homologue groups were di-, tetra-, penta-, and hexa-CBz. A dominance and specific profile of 1,4-DiCBz, PeCBz, and HCBz were characteristic to Halowax 1001, 1099, 1013, 1014, and 1051, while for other formulation the CBzs profile varied, and 1,2,3,4-TeCBz (Halowax 1031) and 1,4-DiCBz (Halowax 1000) predominated. A steric hindrance effect seems to direct a preferential by-side formation of 1,4-DiCBz and further also of PeCBz and HCBz due to relatively elevated temperature as well as duration time of synthesis for five most chlorinated (49-70 Cl%) chloronaphthalene Halowax formulations.

By-side impurities in chloronaphthalene mixtures of the Halowax series: all 19 chlorophenols.

All seven Halowax formulations examined were contaminated with chlorophenols (CPhs). The total CPhs content of the CN mixtures ranged from 1050 to 34,200 ng/g. The absolute concentration of CPhs decreased with an increase of degree of chlorination (% Cl) of the Halowax 1031 (27%), 1000 (35%), 1001 (49%), and 1099 (51%), respectively, but remained relatively constant for higher chlorinated Halowax 1099 (51%), 1013 (54%), 1014 (59%), and 1051 (70%). 2,4,6-TrCPh and 2,4-DiCPh did contaminated all seven Halowax formulations and PeCPh remained undetected (<90 ng/g) only in Halowax 1099, while less frequently occurred 2,6-DiCPh and followed by 3-/4-MoCPh, 2,5-DiCPh, 2-MoCPh, 2,3-DiCPh, and 2,3,4,6-TeCPh. The CPh congeners such as 3,4-DiCPh, 3,5-DiCPh, 2,3,4-TrCPh, 2,3,5-TrCPh, 2,3,6-TrCPh, 2,4,5-TrCPh, 3,4,5-TrCPh, 2,3,4,5-TeCPh, and 2,3,5,6-TeCPh were undetected in any of the Halowax formulation examined. The profile (%) of higher chlorinated CPh homologues like PeCPh in the Halowaxes followed somehow a degree of chlorination (Cl %) of the parent mixture, i.e., increased from 14, 22, 43, 55, 52 to 80%, respectively, but exception was free of PeCPh the Halowax 1099. And contrary, for a lower chlorinated DiCPhs and TrCPhs their relative proportions decreased but again an exception was Halowax 1099. The fingerprint profiles of 2,4,6-TrCPh, 2,4-DiCPh, and PeCPh, which were usually the most abundant congeners varied and were less consisted when compared to the CPh homologue group profiles, and also more depended on the type of technical Halowax mixture. Due to absence of 2,4,5-TrCPh but presence of 2,4-DiCPh, 2,6-DiCPh, 2,4,6-TrCPh, and PeCPh, which, respectively, are indicative congeners for CPhs obtained by alkaline hydrolysis of chlorobenzenes or phenol chlorination using gaseous chlorine, just co-occurrence of phenol impurity in technical naphthalene is suggested as a solely source of CPhs in the Halowax mixtures.

Attempts on freezing the Greylag (Anser anser L.) gander semen.

Semen of Greylag (Anser anser L.) ganders was frozen according to a method previously elaborated by the authors for freezing the White Koluda gander semen. Semen was collected from five to eight Greylag ganders, twice a week during three succeeding reproductive cycles, by dorso-abdominal massage. Semen samples were diluted in the ratio of 1:1 or 2:1 (two parts semen: one part diluent) with EK diluent, supplemented by 6% DMF, equilibrated and pre-frozen to -140 degrees C at a rate 60 degrees C/min, before being transferred into liquid nitrogen container. Semen samples thawed in a water bath of 60 degrees C were used for twice a week insemination in a volume of 200 microl. Three Greylag and three White Koluda geese were involved in frozen-thawed semen fertilizing ability test. The reproductive cycle of wild geese lasts usually about 6-7 weeks. The ejaculate volume (30-140 microl) and sperm concentration (10x10(6) to 150x10(6) ml(-1)) are much lower than these of domestic ganders, but spermatozoa morphology is similar, particularly while compared to 1-year-old White Koluda ganders semen. There are about 90% of live spermatozoa and about 30% of live morphologically normal cells in Greylag gander fresh semen. The Greylag gander spermatozoa susceptibility to cryopreservation procedure is as high as in domestic ganders. Dilution ratio 2:1 resulted in higher number of live spermatozoa, which withstood cryoinjury stress. In relation to fresh semen about 60% of spermatozoa remained intact (on the basis of light microscope examination) in the frozen-thawed semen. Insemination of frozen-thawed semen resulted in 37.5% of fertile eggs in Greylag and 25.0% in White Koluda geese. Low fertility rate was caused by an insufficient number of live normal spermatozoa used for insemination (about three million in every dose).

Effect of age on quality of fresh and frozen-thawed semen in White Italian ganders.

AIM: To comparatively evaluate the fresh semen quality of 1, 2 and 3-yr-old White Italian ganders (Anser anser L.) and the susceptibility of spermatozoa to freezing-thawing procedure. METHODS: Semen was collected by dorso-abdominal massage every 2 days~3 days from three groups of ganders: 1-yr-old (n=11), 2-yr-old (n=7) and 3-yr-old (n=9). In the pooled fresh semen samples, the following parameters were evaluated: the ejaculate volume, the blood or feca contamination and the motility, concentration and morphology of spermatozoa. Sperm motility and morphology were evaluated in the frozen-thawed semen. Semen diluted with EK extender was frozen in straws in a computerized freezing unit with 6 % dimethyl-formamide to -140 deg at a rate 60 deg/min and then transferred into the LN2 container. Straws with semen were thawed in a water bath at 60 deg. RESULTS: The ejaculate volume decreased with the age (0.21 mL for 1-yr-old, 0.18 mL for 2-yr-old and 0.14 mL for 3-yr-old ganders); the sperm concentration increased with the age (327 x 10(6) mL(-1) for 1-yr-old, 431 x 10(6) mL(-1) for 2-yr-old and 547x10(6) mL(-1) for 3-yr-old ganders); the number of live - normal sperm was significantly (P<0.01) lower in the 1-yr-old than that in the 2- and 3-yr-old ganders (26.61 %, 41.54 and 35.9 %, respectively). The percentage of normal cells survived the freezing-thawing process was 37.7 %, 43.3 % and 40.9 % for 1-, 2- and 3-yr-old ganders, respectively. CONCLUSION: Freezing and thawing processes more significantly (P<0.01) affected the motility, viability and morphology of spermatozoa in semen of 1-yr-old ganders in comparison with older males.