A novel canine B‐cell leukaemia cell line. Establishment,characterisation and sensitivity to chemotherapeutics

We established a new B‐cell leukaemia cell line CLB70 from a dog with chronic lymphocytic leukaemia. This cell line is positive for CD20, CD45, CD79a, MHC class II, IgG, IgM; weakly positive for CD21; and negative for CD3, CD4, CD5, CD8, CD14, CD34, CD117. PCR for antigen receptor gene rearrangement (PARR) analysis revealed a biclonal immunoglobulin heavy chain (IgH) gene rearrangement and negative result for TCRγ. Western blot analysis of anti‐ and pro‐apoptotic proteins showed increased expression of Bcl‐2, Mcl‐1, NF‐kB, and Ras, and decreased expression of p53. CLB70 cells grow rapidly in vitro and are tumourigenic in nude mice. The CLB70 line is highly sensitive to doxorubicin, less sensitive to etoposide and imatinib, and resistant to piroxicam, celecoxib and dexamethasone. Our results indicate that CLB70 cells are derived from mature B‐cells and they may be a useful tool for the development of new therapeutic strategies for both dogs and humans.     

Female age as a factor determining the patterns of tree canopy utilisation by the European cherry fruit fly,Rhagoletis cerasi

The European cherry fruit fly, Rhagoletis cerasi is a univoltine, oligophagous pest of cultivated sweet cherries. The pest is closely associated with the canopy of its host tree, and its on-farm behaviour and mobility are determined by the crop structure and spatial arrangement of the farm. However, in spite of its relevance to integrated pest management, little is known about the within-canopy behaviour of R. cerasi, or the role of host and non-host canopy traits in local pest translocations. This paper reports the results of our exploratory studies, which reveal the modulating role of female age on various aspects of R. cerasi on-farm behaviour, such as diurnal and lifetime patterns of canopy utilisation and within-canopy activity, the propensity to undertake local explorative errands and the response to volatiles emanating from host and non-host canopies. The potential implications of our findings on the development of site-optimised IPM and/or pest-resilient agro-landscapes were also discussed.     

UV Light-Assisted Degradation of Methyl Orange,Methylene Blue,Phenol, Salicylic Acid,and Rhodamine B: Photolysis Versus Photocatalyis

Methyl orange (MO), methylene blue (MB), phenol (F), salicylic acid (SA), and rhodamine B (ROD) were used as substrates during the photodegradation experiments in the absence and in the presence of nanostructured Ag/titania-silica. The catalyst was characterized by scanning electron microscopy (SEM), scanning transmission electron microscope high-angle annular dark field (STEM-HAADF), stereological analysis, nitrogen adsorption-desorption, and X-ray photoelectron spectroscopy (XPS) measurements. The results were fitted on pseudo-first and pseudo-second kinetic order models. The film diffusion was also determined. The photolysis degrades MO and F to a greater extent than the photocatalysis. The degradation of SA occurred at the same rate either by photolysis or by photocatalysis. MB was best removed by photocatalysis. With regard to the photocatalysis, the highest rates of film diffusion were obtained for MB, F, and ROD, meaning that these molecules crossed the film to arrive at the catalyst surface more rapidly than the others. For MO and MB, the results followed the pseudo-first-order kinetic model while for SA, F, and ROD, the pseudo-second-order kinetic model was more appropriate.     

Early Cleaved Bovine Embryos Show Reduced Incidence of Chromosomal Aberrations and Higher Developmental Potential on Day 4.5 Post‐Insemination

The present study aimed to investigate whether the timing of the first zygotic cleavage (FZC) influences the speed of embryo development expressed by the total cell count and the rate of chromosomally aberrant embryos. Bovine embryos were produced in vitro and divided into two categories according to the timing of FZC: early cleavers (at 30 hpi; EC) and non‐early cleavers (at 48 hpi; NEC). On day 4.5 pi, embryos were grouped into three classes depending on the number of blastomeres: delayed (<8 BL), normal (8–16 BL) and advanced (>16 BL). We applied fluorescence in situ hybridization (FISH) with probes for bovine chromosomes 6 and X. The only form of chromosomal imbalance observed was mixoploidy [(2n/3n; 2n/4n); 19.9%, 54/271]. Early cleavers were less often chromosomally unbalanced (13.9%, 20/144) than their NEC counterparts (26.7%, 34/127). Among embryos developing at a normal speed, the NEC embryos were more often abnormal (NEC 20/80; EC 10/79; p < 0.05). The advanced embryos were not observed among the NEC category, whereas such embryos from EC category displayed no chromosomal aberrations. The majority of embryos arrested at the 8 BL stage were of NEC category and were carriers of chromosomally abnormal blastomeres. With regard to embryonic sex, we demonstrated that although males dominate among bovine embryos developing in vitro, the incidence of mixoploidy was equal for both sexes. It can be suggested that a good‐quality bovine embryo is usually an early cleaver that develops at higher speed and contains less aberrant cells. The present study also confirmed the usefulness of the FZC as a marker of embryo quality by demonstrating a significantly lower incidence of aberrations in early embryos.     

Proliferation and Apoptosis in Fetal Membranes and Endometrium During Placental Retention in Heavy Draft Mares

Placental retention (retained placenta [RP]) is a serious and common peripartum disease in mares, but the etiology and pathogenesis of RP still remain unclear. The alteration of cell proliferation and apoptosis is considered to be an important factor in RP. Fetal membranes and endometrial biopsies were collected from mares with RP (n = 8) and from control mares (n = 10). The proliferation and apoptosis levels in the chorionic and the endometrial epithelia were assessed by proliferating cell nuclear antigen immunostaining and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay, respectively. The study revealed that there was an insignificant decrease in proliferation and a significant increase in apoptosis in the chorionic epithelium from mares with RP. This result excludes a proliferation imbalance from the possible causes of RP. In the area of the nonpregnant horn of the placenta, proliferation was negatively correlated and apoptosis was positively correlated with the degree of fetomaternal anchorage. It was observed that, in all mares with placental retention, the endometrial epithelium (both luminal and glandular) showed decreased proliferation and increased apoptosis, which may indicate a delay in postpartum uterine regeneration.     

Synthesis and fungistatic activity of new groups of 2,4-dihydroxythiobenzoyl derivatives against phytopathogenic fungi

Twenty-six compounds, derivatives of amides, hydrazines, hydrazides, hydrazones, and semicarbazides, with a 2,4-dihydroxythiobenzoyl moiety, were synthesized from sulfinyl-bis(2,4-dihydroxythiobenzoyl). The compositions and chemical structures of these compounds were confirmed by IR, (1)H NMR, EI-MS, and elemental analysis. Antifungal properties of chemicals under in vitro conditions against five phytopathogenic fungi were estimated. In vivo studies against Erisiphe graminis were also carried out. The compounds N-substituted with an 2,4-dihydroxythiobenzamide group proved to be the most active. N-2-(1-Cinnamylbenzene ester)-2,4-dihydroxythiobenzamide, under in vitro conditions, showed activity at the level of 80-100% development of most pathogens at a concentration of 20 microg/mL and partially at a concentration of 200 microg/mL. For compounds with -HN-NH- or -NH-N= moiety, weak or no fungistatic properties were found at the concentrations studied.     

Effect of sunflower, linseed and fish oils on the production of trans fatty acids in vitro

In vitro anaerobic incubations were used to determine the effect of different oils (LO-linseed, SO-sunflower, FO-fish oil) on trans fatty acid production in rumen fluid and to test if combining of monensin (MON) with the oils affects the interactions on trans fatty acid concentrations in mixed cultures of ruminal microorganisms. Two different sources of rumen fluid were used; the inoculum from the sheep fed hay and barley (80:20%)--the inoculum A and the inoculum from the sheep fed alfalfa and barley (80:20 %)--the inoculum B. The analyses showed that inoculum B contained more short chain fatty acids (SCFA), medium chain fatty acids (MCFA) and saturated fatty acids (SFA) than inoculum A. In contrast, inoculum A contained more unsaturated fatty acids (UFA) than inoculum B. The results show, that the oils affected the biohydrogenation of fatty acids (FA) by increasing the concentration of C18:0 (3-7 times) and trans C18:1 isomers (2-9 times). The concentration of two main intermediates of FA biohydrogenation-- cis 9, trans 11 C18:2 (CLA) and trans 11C18:1 (TVA) were increased with the oils, but FO was more efficient than other plant oils on CLA and TVA production.The monensin treatment had similar effect on FA metabolism as the oil treatment in comparison to unincubated control. The interactions of monensin treatment with the oils were characterized with decrease (LO+MON, SO+MON) or increase (FO+MON) of the proportions of C18:0 and trans C18:1 isomers in comparison to oil treatment.The highest concentrations of two main isomers--CLA,TVA were found in the samples containing fish oil and monensin. In conclusion, fish oil treatment and monensin with fish oil treatment was more efficient than other plant oils in the effect on trans fatty acid production (mainly CLA and TVA) in fermentation fluid in vitro.     

Choice of the optimal method for the isolation of Salmonella from meat- and bone powder designed for industrial feed mixtures

The purpose of this paper was (1) comparison of four multi-step methods used for Salmonella isolation from meat- and bone powder; (2) elaboration of a new sensitive method of Salmonella isolation from this product; (3) evaluation of a new solid selective medium (BxLH) described by the authors for Salmonella isolation in comparison to brilliant green agar (BGE) according to Edel and Kampelmacher. The study was carried out on 173 meat- and bone powder samples naturally contaminated with Salmonella oranienburg. The samples were examined for the Salmonella presence by means of four compared methods (Methods 1 to 4). The new method of isolation proposed by the authors (Method 3) proved to be the most effective among all compared for Salmonella recovery. It seems that the superiority of Method 3 in comparison to the other applied was a result of, (1) homogenization of the investigated samples in distilled water before preincubation followed by maintenance at room temperature for 2–4 h; (2) the use of a new selective BxLH agar; (3) the use of multiple plating after selective enrichment. The BxLH medium was shown to be more suitable for Salmonella isolation than BGE agar because of more efficient inhibition of other bacterial growth with simultaneously abundant growth of the Salmonella organisms. The additional advantage offered by BxLH agar was the fact that lactose-positive salmonellas grow as typical representatives of this genus. This enables their identification, in contrast to the situation when lactose containing media are used, where the colonies of such salmonellas are similar to the colonies of, for example, Escherichia coli.     

Identification of a new self-incompatibility allele (S31) in a Hungarian almond cultivar and its reliable detection

Self-incompatibility of almond [Prunus dulcis (Mill.) D.A. Webb] is controlled by the S-locus with 30 described allelic variants. In this study, PCR amplification, cloning and DNA sequence analysis revealed a new S-RNase allele in a Hungarian cultivar, ‘Tétényi bőtermő’. This new allele was labelled as S₃₁. Since S₃₁ is characterized by almost identical intron sizes as S₉, consensus PCR was not successful in discrimination of the alleles, even if fluorescently labelled fragments were sized on an automated sequencer. Therefore, an allele-specific forward primer (PdS31-F) was designed to anneal selectively within the second intron of the S₃₁-RNase gene and used in combination with the EM-PC3consRD consensus primer. This allowed for the successful discrimination of S₃₁ from S₉. The PdS31-F primer and allele-specific PCR in general might be useful in the identification of different alleles with matching intron sizes that might occur during screening for S-alleles in a more diverse population, e.g. local cultivars from Central Europe to Asia.     

Stress and its influence on reproduction in pigs: a review

The manifestations of stress, defined as a biological response to an event that the individual perceives as a threat to its homeostasis, are commonly linked to enhanced activity of the hypothalamo-pituitary-adrenal (HPA) axis and the activation of the sympathetic adreno-medullary (SA) system. Activation of the HPA system results in the secretion of peptides from the hypothalamus, principally corticotropin releasing hormone (CRH), which stimulates the release of adrenocorticotropic hormone (ACTH) and beta-endorphin. ACTH induces the secretion of corticosteroids from the adrenal cortex, which can be seen in pigs exposed to acute physical and/or psychological stressors. The present paper is a review of studies on the influence of stressors on reproduction in pigs. The effects of stress on reproduction depend on the critical timing of stress, the genetic predisposition to stress, and the type of stress. The effect of stress on reproduction is also influenced by the duration of the responses induced by various stressors. Prolonged or chronic stress usually results in inhibition of reproduction, while the effects of transient or acute stress in certain cases is stimulatory (e.g. anoestrus), but in most cases is of impairment for reproduction. Most sensitive of the reproductive process are ovulation, expression of sexual behaviour and implantation of the embryo, since they are directly controlled by the neuroendocrine system.