Nitric oxide induces apoptosis in bovine luteal cells

We previously showed in in vivo and in vitro studies that nitric oxide (NO) is engaged in luteolysis in cattle. Nitric oxide produced locally in the bovine corpus luteum (CL) inhibits progesterone (P4) synthesis and is suggested to be a component of the luteolytic cascade induced by uterine prostaglandin (PG) F2alpha. In the present study, the molecular mechanisms of NO action during structural luteolysis were studied in cultured bovine luteal cells (Days 15-17 of the estrous cycle). The effects of the NO donor (NONOate; 10(-4)M) on DNA fragmentation, cell viability, P4 production and caspase-3 activity were compared with those of PGF2alpha (10(-6)M). Moreover, mobilization of intracellular calcium [Ca2+]i and gene expressions of Fas-L, Fas, bcl-2, bax, and caspase-3 in the cells were determined by semi-quantitative RT-PCR after NONOate treatment. Caspase-3 activity was examined calorimetrically. Contrary to PGF2alpha NONOate decreased cell viability. DNA fragmentation after NONOate treatment increased by more than with PGF22alpha. NONOate increased mobilization of [Ca2+]i in the cells. Although the NO donor did not affect Fas-L and bcl-2 gene expression, it stimulated Fas and bax mRNA and caspase-3 expression. The ratio of bcl-2 to bax mRNA level decreased in the cells treated with NONOate. Moreover, NONOate stimulated caspase-3 activity more effectively than PGF2alpha. The overall results suggest that NO is a luteolytic factor that plays a crucial role in regulation of the estrous cycle in structural luteolysis by inducing apoptosis of luteal cells in cattle.     

Effect of steroidal and non-steroidal anti-inflammatory drugs in combination with long-acting oxytetracycline on non-specific immunity of calves suffering from enzootic bronchopneumonia

The aim of this paper was to compare the effect of flumethasone and meloxicam in combination with oxytetracycline on clinical and immunological parameters of calves suffering from enzootic bronchopneumonia. The study was performed on 30 Black-and-White Lowland Breed calves with clinical signs of enzootic bronchopneumonia divided randomly into three equal groups and, respectively, treated with-Group I: oxytetracycline and meloxicam; Group II: oxytetracycline and flumethasone; Group III (control): oxytetracycline only. Treatment of calves with the combination of oxytetracycline and meloxicam (Group I) caused a significantly faster, in comparison to other groups, improvement in the clinical illness index score (CIIS: cough, nasal discharge, dyspnea, depression and anorexia) and a faster normalization of body temperature. A slow decrease in white blood cell (WBC) count, the number of neutrophils, MID (mixed number of monocytes, eosinophils and basophils) and in the individual number of monocytes (CD14/CD45 positive cells) was observed in Groups I and III. In the blood of the calves which received oxytetracycline and flumethasone (Group II), leukocytosis, neutrophilia and monocytosis with concomitant lymphopenia and a low number of T cells (CD2+) was observed. Moreover, the calves treated with flumethasone exhibited a decrease in gamma-globulin concentration, and phagocytic parameters. Both drugs, flumethasone and meloxicam slightly decreased tumor necrosis factor (TNF) but meloxicam slightly increased the levels of interferon (IFN) in sera and in bronchoalveolar lavages (BALs). These results suggest that the combination of meloxicam with an antibiotic in calves suffering from enzootic bronchopneumonia is superior to the antibiotic alone and also to the combination of the antibiotic with flumethasone.     

Assessment of soil texture class on agricultural fields using ECa,Amber NDVI,and topographic properties

Soil texture (ST) is relatively stable over time, although it may change due to erosion, clay eluviation, and other processes. Soil texture affects soil quality, productivity and management. Therefore, indirect, accurate methods for assessing of soil texture classes (STCs) are needed in agricultural practice. A study was performed on four production fields in northern and central Poland to compare the fitting performance of STC models based on apparent electrical conductivity (ECa), topographic properties (elevation, slope gradient and wetness index) and Amber NDVI measurements. One common and accurate indicator of STCs was not found for all study fields. On average, ECa was most accurate in indicating areas of different STCs within the fields, but it tended to overestimate the size of sandy areas on loamy fields and vice versa. The accuracy of STC assessment using ECa measurements may be biased due to imperfect soil drainage, high elevations, which increase evaporation and STC variation with depth. STC assessment using Amber NDVI measurements may be useful, particularly on flat and sandy fields, but the results are affected by the same factors as ECa, and additionally by crop growth stages and by the weather conditions in the period preceding the measurements. Despite the good quantitative results of the STC assessment by elevation (one field) and by the topographic wetness index (another field), both terrain attributes failed to accurately indicate the distribution of some STC areas within each field. Therefore, in landscapes developed from deposits of the last glaciation relevant ST differences might not sufficiently be detected by the analysis of terrain attributes alone. The selection of STC predictors and evaluation of the assessment quality must consider both the quantitative indicators such as correlation and determination coefficients describing relationships between ST and ECa, NDVI and topography and percentage of a field area with accurately indicated STC and the distribution of areas with different STCs within a field. The use of ECa, NDVI values, and topographic properties for STC assessment may be useful in reducing costs of soil sampling and analysis, but cannot replace it.     

Conjugated linoleic acid regulates phosphorylation of PPARγ by modulation of ERK 1/2 and p38 signaling in human macrophages/fatty acid-laden macrophages

Stimulation of macrophages by a variety fatty acids causes activation of MAP kinases (MAPKs). The consequences arising from down-regulation of MAPKs may be a limitation in the activity of PPARγ, which is modulated by a modification catalyzed by these kinases. Phosphorylation of MAP kinases-ERK1/2 and p38 as well as PPARγ was determined by real-time polymerase chain reaction and Western blotting in human macrophages cultured with conjugated linoleic acids (CLAs). We demonstrated that CLA isomers alter MAP kinase phosphorylation and PPARγ activation. Phosphorylation of ERK1/2 was diminished in cells cultivated with cis-9,trans-11 CLA, whereas phosphorylation of p38 was reduced by trans-10,cis-12 CLA. PPARγ was phosphorylated mainly by ERK1/2, and consequently, PPARγ phosphorylation was suppressed mainly by cis-9,trans-11 isomer. In human adipocytes, cis-9,trans-11 C 18:2 raised the activation of PPAR and several of its downstream target genes. We suggest that a similar process may also occur in human macrophages.     

Influence of water salinity on the survival of embryos and growth of the sichel larvae Pelecus cultratus (L.) under controlled conditions

The purpose of this study was to analyse the influence of water salinity on the incubation of eggs and growth of the larvae of sichel Pelecus cultratus (L.) under controlled conditions. The following water salinity was considered: 3, 6, 9, 12, 15 and 0 ppt for the control. The eggs incubation time depended on the water salinity, lasting from 56.3°D [9 ppt] to 107.7°D [0 ppt]. The higher water salinity, shorter the incubation time. The highest survivability of larvae was obtained in water of the salinity equal 3 ppt and in freshwater, where respectively 87% and 84% of the larvae survived. No hatched larvae were obtained in water of the salinity of 12 and 15 ppt, as all embryos died during the experiment. Larvae were reared up for 21 days. The final body weight, depending on the water salinity, reached from 43.5 mg [9 ppt] to 74.6 mg [3 ppt], at the respective body lengths of 19.3 mm and 23.2 mm. The highest body gains and lengths of larvae were observed in water of the salinity of 3 ppt. The lowest survivability of sichel larvae occurred in water of the salinity 9 ppt, where it was only 53.2%. The survivability of larvae in freshwater and water of the salinity of 3 ppt was quite similar: 96.2% and 95.6% respectively. Water salinity degree of 12 and 15 ppt appeared to be lethal for both embryos and larvae of the sichel.     

Genetic diversity of carrot (<Emphasis Type="Italic">Daucus carota</Emphasis> L.) cultivars revealed by analysis of SSR loci

Polymorphism of simple sequence repeat (SSR) loci was assessed in a collection of 88 carrot (Daucus carota L. subsp. sativus Hoffm.) accessions. The collection comprised cultivars and landraces mainly from Asia, Europe, and North America. Plants were grown in the glasshouse and characterized for root color and shape. Thirty SSR loci were fully characterized using parameters derived from allele frequencies, i.e. the number of total, effective and rare alleles, the observed and expected heterozygosity, and fixation index. Using a Bayesian approach, two clusters of 17 and 61 accessions were distinguished, which comprised the Asian and Western type accessions, respectively. Genetic diversity of the Asian gene pool was higher than that of the Western gene pool. The results of SSR analysis were supported by morphological characterization, and are congruent with current knowledge on the history of carrot domestication and breeding.     

The effect of different dietary ratios of lysine,arginine and methionine on biochemical parameters and hormone secretion in turkeys

We postulated that the use of optimal levels and proportions of Lys, Arg and Met in compound feed allows for optimal exploitation of the growth potential of contemporary slaughter turkey hybrids and reduces metabolic disorders. The aim of the study was to determine the effect of different proportions of Lys, Arg and Met in diets whose Lys content is in accordance with NRC recommendations, that is a low level, on selected parameters of protein, lipid and carbohydrate metabolism and on hormone secretion in turkeys. The lowest Arg content (90% Lys) in the diet resulted in an increase in plasma total cholesterol levels in the turkeys as compared to higher Arg content (100% or 110% of Lys), (2.50 vs. 2.09 vs. 1.83). Plasma HDL and creatinine concentration increased in turkeys fed diets with higher Arg content (100% and 110% Lys) compared to turkeys receiving the diet with the lowest Arg content (90% Lys). Compared to turkeys receiving the lowest and intermediate Arg content (90% and 100% Lys), the diet with the highest content of this AA (110% Lys) resulted in an increase in the plasma T4 level (71.21 vs. 86.60 vs. 128.2). The varied Arg and Met levels relative to Lys did not affect the secretion of neurotransmitters or hormones regulating glucose metabolism. At low levels of Met in the diet, an decrease in Arg relative to Lys from 100% to 90% caused a growth depression of turkeys (10.68 vs. 10.21 kg), which was not noted in the case of the higher Met content. When using the Lys level recommended by NRC in the turkey diet, the optimal Arg level is 100% and Met is 45% compared to Lys.     

A new classification of pre‐ovulatory oocyte maturation stages in pikeperch,Sander lucioperca (L.), and its application during artificial reproduction

This study aimed at improving the reproduction effectiveness and synchronization of ovulation in the pikeperch, Sander lucioperca (L.), during induced spawning, which is one of the main bottlenecks in the aquaculture of this species. For this purpose, a new categorization of maturation stages in pre‐ovulatory oocytes was applied. It is generally based on two morphological indicators: germinal vesicle migration or its breakdown (GVBD) and different oil droplet coalescence rates. This categorization covered seven stages (from I to VII) – from the end of vitellogenesis to ovulation. The categorization was verified by controlled reproduction with the use of hormonal stimulation (500 IU of hCG per kg of female body weight) and low spawning temperature (12 °C), which extended the latency time. In addition, some morphological indicators (pseudo‐gonadosomatic index, Fulton's condition coefficient) of females were calculated in order to determine their usability in determining the maturation stage. However, these indicators proved to be ineffective for this purpose, further highlighting the need to determine the maturational stages in pre‐ovulatory oocytes to synchronize ovulation in pikeperch. During the experiment, ovulation seemed to be synchronized among the experimental treatments. Statistical differences were found in terms of latency time between experimental groups at different maturity stages (II – 78–98 h; III – 57–78 h; IV – 48–58 h; V – 32–49 h; VI – 5–30 h) according to the proposed classification. This classification and the results presented in the study significantly improved the synchronization of ovulation, which may positively affect the effectiveness of pikeperch production under controlled conditions.     

Rearing river lamprey <Emphasis Type="Italic">Lampetra fluviatilis</Emphasis> (L.) larvae under controlled conditions as a tool for restitution of endangered populations

A method for actively protecting river lamprey and increasing the number of river lamprey populations is presented as protocol of rearing stocking material. The objective of the study was to accommodate lamprey larvae during the most critical life period under controlled conditions until they reach a sufficient size, which will increase their chances to survive in the natural environment. The eggs obtained from wild breeders were incubated in Weiss’s jars at the water temperature of 12 °C. The larvae were reared in tanks set up in closed recirculating aquaculture systems. During the rearing period, the water temperature was 20 °C and the density of larvae was 150 individuals per 1 dm² of bottom. In the initial rearing phase, the main focus was on the type of feeds and the nourishment method. It was determined that the most suitable feed type was Artemia nauplii combined with Hikari dry food. Additionally, the type of substrate was tested under controlled conditions. In this case, the optimum one was composed of sand of the grain size between 0.3 and 0.5 mm. The experiment proved that it is possible to successfully conduct river lamprey larvae rearing under controlled conditions. The growth rates are comparable to or better than those achieved under natural conditions. After 30 days of the rearing period, the lamprey larvae were 25 mm long. The results create a possibility to develop a technology for the restitution of river lamprey and other lamprey species, which are endangered throughout the world.