Relationships between innovative and traditional parameters to investigate semen quality in pigs

Semen quality assessment represents a fundamental step for obtaining successful artificial insemination (AI). In commercial settings, the semen employed for AI should be of high quality but traditional semen quality estimates are not sufficiently sensitive to discriminate between differences among samples in terms of fertilising ability. Therefore, more discriminative sperm characteristics need to be identified in order to better predict fertility outcome. In the present study, a series of molecular aspects of semen, represented by heat shock proteins, oxidative stress status, antioxidant potential and tumour necrosis factor (TNF)-alpha were evaluated and analysed. Several relationships between traditional and investigated molecular semen quality estimates were found by using a multivariate analysis approach. Tumour necrosis factor (TNF)-alpha was identified in boar seminal plasma resulting in positive correlations with several sperm quality aspects and particularly with motility. The protective roles of antioxidant molecules and heat shock proteins have been demonstrated confirming the data previously published in the literature.     

Antigenic Relationship of Brucella ovis and Brucella melitensis

Immune sera were prepared in rabbits by the injection of living and acetone-killed cells of Brucella ovis and smooth and rough B. melitensis. The use of whole-cell antigens in agglutination and agglutinin-absorption tests revealed little relationship between B. ovis and smooth B. melitensis, although there was extensive cross-agglutination between B. ovis and rough B. melitensis. The use of water-soluble antigens prepared from ultrasonically treated cells of the three strains revealed extensive cross-reactions in indirect hemagglutination, agar gel precipitation, and immunoelectrophoresis tests, as well as in allergic skin tests in rabbits. The most definitive results were obtained with the immunoelectrophoresis technique. B. ovis antigen produced at least 11 lines with its homologous serum. All were removed by absorption of the serum with rough B. melitensis antigen. All but three were removed by absorption with smooth B. melitensis antigen. Smooth B. melitensis antigen produced 11 lines with its homologous serum, and all but 3 were removed by absorption with B. ovis antigen. Rough B. melitensis produced nine lines with its homologous serum, and eight were removed by B. ovis antigen. The extensive cross-reactions between soluble antigens of B. ovis and B. melitensis are added evidence that B. ovis belongs in the genus Brucella.     

Homologies of Deoxyribonucleic Acids from Brucella ovis, Canine Abortion Organisms, and other Brucella Species

The bacterium that causes canine abortion has polynucleotide sequences similar, in deoxyribonucleic acid (DNA)-DNA homology studies, to those of Brucella suis and, by inference from previous data, those of B. abortus and B. melitensis as well as B. neotomae. Therefore, the organism causing canine abortion appears to be a member of the genus Brucella. DNA preparations from Serratia marcescens, Alcaligenes faecalis, and Bordetella bronchiseptica, 58, 62, and 66 mole% guanine plus cytosine, respectively, do not have detectable polynucleotide sequence homologies with B. suis DNA which is 56 mole% guanine plus cytosine. B. ovis DNA lacks some of the polynucleotide sequences present in B. suis DNA and appears to be a deletion mutant. However, a large proportion of B. ovis polynucleotides are similar to those of other Brucella species, which supports the inclusion of B. ovis in the genus.     

Relationships between basic life history parameters and population size

We consider an ideal population with a stable age composition changing according Lotka equation. Additional assumptions are made concerning the constancy of population size, independence of specific mortality rate on age, and linear dependence of female fecundity on its weight. A relationship has been obtained [formula: see text] where N0 is initial numbers of a generation, N[alpha, omega] is total numbers of the mature part of the population, w[alpha, omega] is a mean weight of a mature individual, s is sex ratio, c is specific fecundity (per unit of weight) and l0 is the probability of larval surviving. The growth of an individual is described by the Bertalanffy function. Methods of calculation of life history parameters are discussed. A method is proposed to calculate the age of maturity (alpha) and at the end (omega) of the reproduction period as first and second inflection points of the growth rate curve. Based upon data on development of 27 populations of several species of fishes of inland waters of Russia the following relationship have been obtained: [formula: see text] for populations with [formula: see text] < or = 100 g, [formula: see text] for populations with [formula: see text] > 100 g, and [formula: see text] for all populations.     

Fatty acid makeup of various Brucella species and its relationship to the culture medium]

Gas chromatographic method was applied to the study of the fatty acid composition (in Br. melitensis, Br. abortus, Br. suis, and Br. ovis strains. Fatty acid composition was similar in the mentioned brucellae species, except Br. suis No. 1330 significantly differing by this sign. Methyleneoctadecanoic acid content was considerably elevated, and that of octadecenoic -- reduced in brucellae grown on liver agar with the addition of serum and on meat-peptone agar in comparison with brucellae grown on liver agar; apparently this represents one of the mechanisms of the microorganism adaptation to the less favourable conditions of the nutrient medium. Passage of Br. ovis strain through the guinea pig organism led to the appearance of brucellae forming two types of colonies when grown on liver agar with the addition of serum. The fatty acid composition of brucellae forming small transparent colonies was the same as that of the initial culture with the prevalence of methyleneoctadecanoic acid; as to brucellae with larger colonies with irregular margin and nontransparent centre of the colony--octadecenoic acid prevailed in their fatty acid composition, i.e. their composition was similar to such in brucellae of the melitensis and abortus species grown on liver agar.     

Studies of Antigens for Complement Fixation and Gel Diffusion Tests in the Diagnosis of Infections Caused by Brucella ovis and Other Brucella

Sonically treated and saline-extracted antigens of Brucella ovis, B. canis, B. abortus, and B. melitensis were compared in gel diffusion, complement fixation, and serum absorption tests. All the sonically extracted antigens showed cross-reactions with sera from animals infected or immunized with these species, whereas the saline-extracted antigens were specific for the surface of the rough or smooth colonial phase of the species or strain. The saline-extracted antigens of B. ovis and B. melitensis were both eluted as a single peak in the void volume by Sephadex G-200 column chromatography, in gel diffusion had staining characteristics of lipoproteins, but in immunoelectrophoresis showed distinct mobility patterns. Serological activity for both gel diffusion and complement fixation tests was demonstrated in the immunoglobulin G-containing fraction of sera taken from sheep 12 to 412 days after infection with B. ovis. The gel diffusion test with saline extract of B. ovis is as sensitive as the complement fixation test for the diagnosis of ram epididymitis and is more practical.     

Cholesterol,ganglioside GM1 and class A scavenger receptor contribute to infection by Brucella ovis and Brucella canis in murine macrophages

The establishment of infection by Brucella ovis and Brucella canis in J774.A1 macrophages was found to be dependent upon cholesterol and ganglioside GM₁, two components of lipid rafts. This process also required a class A scavenger receptor of macrophages, and was not inhibited by smooth and rough lipopolysaccharides from Brucella spp. In response to infection, both bacteria induced a weak degree of macrophage activation. These results demonstrate that B. ovis and B. canis use cell surface receptors common to smooth Brucella spp. for macrophage infection, thus limiting macrophage activation and favouring intracellular multiplication and/or the survival of both bacteria.     

Relationship between Semenogelins bound to human sperm and other semen parameters and pregnancy outcomes

Background

Semenogelins (SEMGs) are major components of human seminal vesicle secretions. Due to SEMG’s sperm-motility inhibitor, a significant negative correlation between sperm motility and the proportion of SEMG-bound spermatozoa (SEMG+) was found in asthenozoospermic patients. SEMGs also show intrinsic inhibitory capability for sperm capacitation; however, studies on actual clinical specimens have not been conducted.

Methods

To reveal the relationship between SEMGs and the fertilizing capacity of sperm from male infertile patients who are not restricted to asthenozoospermia, we measured the proportion of SEMG+ in the spermatozoa of 142 male infertile patients. The pregnancy outcomes in partners of these patients were retrospectively analyzed using questionnaires.

Results

Among examined semen parameters, only the total SEMG-unbound sperm count showed a tendency to be different between the spontaneous pregnancy or intra-uterine-insemination-pregnancy groups and in-vitro-fertilization- or intracytoplasmic-sperm-injection-pregnancy groups. It was elevated in the former group, which includes patients who used in vivo fertilization.

Conclusions

The total SEMG-unbound sperm count would be a relevant parameter for in vivo fertilization. This result suggests that SEMGs inhibit ectopic capacitation before sperm reach the fertilization site and that the number of total SEMG-unbound sperm is a parameter directly linked to the possibility of in vivo fertilization.

     

Relationships between morphological parameters in birds with different flying habits

1. Data on morphological and physiological parameters from 346 species of birds were collected from diverse sources. 2. The observed relationships among some pairs of variables showed differences between the five flight styles into which the studied birds were classified. 3. The characteristic kind of flight for each species is related to four easily obtainable non-invasive parameters: body weight (BW), wing area (WA), wing span (WS) and boyd length (BL). 4. Wing-loading index (BW2/3/WA) gives the most effective variable to describe the flying habits of any species.     

Relationships between culture density and the composition of three floating aquatic macrophytes

Eichhornia crassipes (Mart.) Solms, Pistia stratiotes (L.), and Lemna minor (L.) were cultured at four different densities each and analyzed for cell-wall fraction, crude protein, total available carbohydrate and ash. Cell-wall fraction increased and crude protein content decreased as density increased in Eichhornia and Pistia cultures. The cell-wall and crude protein content of Lemna did not change with increasing culture density. Differences in the trends of cell-wall and crude protein content of the three plants at increasing culture densities appear to be related to differences in growth form. There was no difference in the total available carbohydrate or ash content of the three species at different culture densities.     

Relationships between unconjugated and sulphated steroids in porcine primary Leydig cell culture

Steroidogenesis in immature porcine Leydig cells was investigated in primary culture at 48-84 h under basal conditions and in the presence of hCG. The basal accumulation of unconjugated steroids was close to linear only during the first 4 h of study, whereas the sulphate-conjugated steroids accumulated essentially linearly over the 36 h experimental period. At the last time point, 95% of the steroids measured were sulphated. Stimulation with hCG (1 ng/ml) led to a still more pronounced sulphate conjugation, and approx 99% of the steroids measured were sulphated at 36 h. Under maximal stimulation with hCG (100 ng/ml) the sulphates accounted for 74% of the total steroids measured at 36 h. Testosterone, androstenedione, dehydroepiandrosterone, 5-androstene-3 beta, 17 beta-diol and estrone were usually quantitatively the most important unconjugated steroids, and sulphated dehydroepiandrosterone, estrone, testosterone and 5-androstene-3 beta, 17 beta-diol were the most important steroid sulphates, especially following maximal stimulation of the cultures. These data emphasize the importance of steroid sulphates in porcine testicular steroid metabolism. Under stimulation with hCG, there was a rapid response in testicular steroidogenesis, initially seen as a rapid increase in the secretion of unconjugated and sulphated steroids. At approx 4-12 h, the rate of sulphate conjugation appeared to reach or even to exceed that of steroid biosynthesis, which lead to stabilisation or a decrease in the concentrations of unconjugated steroids. Only high doses of hCG, 10-100 ng/ml, were then able to lead to a net accumulation of unconjugated steroids, at 24-36 h of incubation with hCG.     

Extraction of membrane antigens from Brucella ovis and an assessment of their serological activity by immunoblotting

The efficacy and selectivity of chaotropic and phase-partitioning procedures for the extraction of membrane proteins from Brucella ovis were compared with a standard Sarkosyl method. Major group 1, 2 and 3 outer-membrane proteins (OMPs) of B. ovis stained by Coomassie blue in SDS-PAGE gels had, respectively, apparent molecular masses of 81/82 kDa, 39-41 kDa and 30-32 kDa. The presence of these bands in the Sarkosyl extract of total membrane vesicles (TMVs) indicate that the procedure failed to selectively solubilize only inner-membrane proteins (IMPs). SDS-PAGE analyses also revealed the presence of OMPs and other additional bands following extraction of B. ovis TMVs by butanol phase-partitioning or with extraction solutions based on the chaotropic reagents potassium thiocyanate (KSCN), sodium salicylate (SSC) and lithium acetate (LAE). OMPs are therefore not selectively extracted by any one of these procedures. Based on the number and staining intensity of extracted membrane-associated polypeptides, the efficacy of different extraction procedures could be graded in decreasing order as follows: KSCN, SSC, butanol and LAE. Both butanol and SSC were particularly effective in extracting group 3 OMPs. Sera from chronic excretor rams were used to identify zones of seroreactivity in immunoblots. Essentially, two reactivity patterns were seen: strong antibody binding against polypeptides in zones A (46-85 kDa), C (28-32 kDa) and D (18-22 kDa) in one, and additional reactivity against zones B (34-44 kDa) and E (13-18 kDa) polypeptides in the other.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationships between morphometry,geographic location and water quality parameters of European lakes

I addressed the question how lake and catchment morphometry influences water chemistry and water quality over a large scale of European lakes, and developed the regression equations between most closely related morphometric and water quality indices. I analysed the data of 1,337 lakes included in the European Environment Agency (EEA) database, carrying out separate analyses for three basic lake types: large lakes (area ≥100 km², 138 lakes), shallow lakes (mean depth ≤3 m, 153 lakes) and large and shallow lakes (area ≥100 km² and mean depth ≤8 m, 35 lakes). The study revealed that in Europe, the lakes towards North are larger but shallower and have smaller catchment areas than the southern lakes; lakes at higher altitudes are deeper and smaller and have smaller catchment areas than the lowland lakes. Larger lakes have generally larger catchment areas and bigger volumes, and they are deeper than smaller lakes, but the relative depth decreases with increasing surface area. The lakes at higher latitudes have lower alkalinity, pH and conductivity, and also lower concentrations of nitrogen and phosphorus while the concentration of organic matter is higher. In the lakes at higher altitudes, the concentration of organic matter and nutrient contents are lower and water is more transparent than in lowland lakes. In larger lakes with larger catchment area, the alkalinity, pH, conductivity and the concentrations of nutrients and organic matter are generally higher than in smaller lakes with smaller catchments. If the lake is deep and/or its residence time is long, the water is more transparent and the concentrations of chlorophyll a, organic matter and nutrients are lower than in shallower lakes with shorter residence times. The larger the catchment area is with respect to lake depth, area and volume, the lower is the water transparency and the higher are the concentrations of the nutrients, organic matter and chlorophyll as well as pH, alkalinity and conductivity. The links between lake water quality and morphometry become stronger towards large and shallow lakes. Along the decreasing gradients of latitude, altitude and relative depth, the present phosphorus concentration and its deviation from the reference concentration increases.     

Relationships between mixis in Brachionus plicatilis and preconditioning of culture medium by crowding

Several experiments with Brachionus plicatilis have been conducted to test the existence of chemical-mediated induction of mixis. In a first experimental set, bioassays were used to test relationships between preconditioning of culture medium to high population density and the occurrence of mixis in mass cultures with these media. The results show that a preconditioned medium has inducing properties that are comparable to the crowding effect.In order to isolate the effect on mixis of the preconditioning, we also carried out an experiment involving individual cultures. Isolated individuals of B. plicatilis, CU strain, placed in 1 ml of medium renewed daily showed no mixis, but mixis was induced when medium preconditioned to high density was used following the same experimental procedure.     

Relation between various cytological parameters of cultured irradiated thymus cells

Bat thymocytes were incubated in a medium with isologous serum. During the first hours following gamma-irradiation alongside with the nuclear pyknosis a dose-dependent decrease in the ability of formation of rosettes with autologous erythrocytes and inhibition of mitotic activity were noted. A conclusion is made of a possible relationship between postirradiation disturbances of membranes and thymocyte nuclei.     

Relationships between uterine culture, cytology and pregnancy rates in a Thoroughbred practice

Endometrial cytology and culture specimens (n=2123) were collected concurrently with a guarded uterine culture instrument from 970 mares (738 barren, 1230 foaling and 155 maiden mares) during three breeding seasons (2001-2004). Results were compared to the 28-d pregnancy rate for the cycle from which the samples were taken. Cytological smears were evaluated for inflammation at x100 and graded as: not inflammatory (0-2 neutrophils/field), moderate inflammation (2-5 neutrophils/field), severe inflammation (>5 neutrophils/field), or hypocellular (scant epithelial cells and no neutrophils). Uterine culture swabs were plated within 6h, incubated for 72 h and results determined at 24, 48, and 72 h. Approximately, 20% (n=423) cytology samples were positive for inflammation (>2 neutrophils), whereas approximately 11% (n=231) of cultures had microorganisms recovered. A majority (64%) of the positive cultures (147/231) had inflammation on cytology smears. Streptococcus equi subsp. zooepidemicus was associated with more positive cytology results than coliforms (P<0.01). Mares with positive cytology or culture had lower pregnancy rates than mares with normal findings (P<0.01). Lowest pregnancy rates were recorded for mares with severe endometrial inflammation (21%, versus moderate inflammation 48%). Isolation of a microorganism from mares with endometrial inflammation was not associated with a further reduction in pregnancy rates. In barren, foaling and maiden mares, cytology was positive in 28, 17, and 5%, respectively, and culture was positive in 12.2, 11.1, and 3.2%. Foaling and maiden mares had higher pregnancy rates than barren mares (62, 69, and 44%, respectively, P<0.001). In conclusion, a positive cytology was twice as common as a positive culture, and isolation of microorganisms was associated with reduced pregnancy rates, even in the apparent absence of inflammation.