Morphological and molecular identification and PCR amplification to determine the toxigenic potential of Fusarium spp. isolated from maize ears in southern Poland

The average amount of precipitation in spring and summer 2010 and 2011 coupled with relatively high temperatures caused massive Fusarium spp. infection of maize and yield losses in southern Poland. In order to examine the cause of this disease outbreak, Fusarium spp. were isolated and fungal strains were identified based on morphological characters and species-specific PCR assays. A total of 200 maize samples were processed, resulting in the obtention of 71 strains, which belonged to five Fusarium species, F. poae being the predominant one (74.56%). Other isolates were identified as F. graminearum, F. oxysporum, F. verticillioides and F. proliferatum. PCR-based detection of mycotoxin-synthesis-pathway genes was also used to determine the potential of the analyzed strains to produce trichothecenes (DON and NIV) and fumonisins (FUM). Only 14 isolates revealed the potential to produce DON (11 strains) and FUM (3 strains). HPLC analyses of grain samples revealed the presence of DON only – other mycotoxins were not detected. Moreover, 57.1% of potentially mycotoxin-producing isolates indicated the toxicity in a biological test.     

Rhamnolipids Increase the Phytotoxicity of Diesel Oil Towards Four Common Plant Species in a Terrestrial Environment

The study focused on assessing the influence of rhamnolipids on the phytotoxicity of diesel oil-contaminated soil samples. Tests evaluating the seed germination and growth inhibition of four terrestrial plant species (alfalfa, sorghum, mustard and cuckooflower) were carried out at different rhamnolipid concentrations (ranging from 0 to 1.200?mg/kg of wet soil). The experiments were performed in soil samples with a different diesel oil content (ranging from 0 to 25?ml/kg of wet soil). It was observed that the sole presence of rhamnolipids may be phytotoxic at various levels, which is especially notable for sorghum (the germination index decreased to 41?%). The addition of rhamnolipids to diesel oil-contaminated soil samples contributed to a significant increase of their phytotoxicity. The most toxic effect was observed after a rhamnolipid-supplemented diesel oil biodegradation, carried out with the use of a hydrocarbon-degrading bacteria consortium. The supplemention of rhamnolipids (600?mg/kg of wet soil) resulted in a decrease of seed germination of all studied plant species and an inhibition of microbial activity, which was measured by the 2,3,5-triphenyltetrazolium chloride tests. These findings indicate that the presence of rhamnolipids may considerably increase the phytotoxicity of diesel oil. Therefore, their use at high concentrations, during in situ bioremediation processes, should be avoided in a terrestrial environment.     

Mechanistic insights into furan formation in Maillard model systems

Furan has recently received considerable attention as a possibly carcinogenic compound occurring in thermally processed foods. Although several food constituents have been identified as furan precursors, multiple formation pathways remain unclear. Therefore, the mechanisms of furan formation in Maillard model systems were studied by means of the carbon module labeling (CAMOLA) technique. Under both roasting and pressure-cooking conditions, furan was formed from glucose via the intact skeleton, and its formation pathways from glucose alone were not amino acid-dependent. However, some amino acids, especially alanine and serine, did influence the furan production by providing an additional formation pathway. Furthermore, most amino acids enhanced the furan production from glucose. Roasting conditions produced 25-100 times higher amounts of furan as compared to pressure-cooking conditions. Surprisingly, in the alanine/glucose model systems, the relative importance of furan production from glucose alone and from the combination of a glucose-derived and an alanine-derived fragment changed completely over a limited time course of 60 min.     

Comparison of P-wave dispersion in healthy dogs,dogs with chronic valvular disease and dogs with disturbances of supraventricular conduction

Background

P-wave dispersion (P_d) is a new ECG index used in human cardiology and veterinary medicine. It is defined as the difference between the maximum and the minimum P-wave duration recorded from multiple different ECG leads. So far no studies were performed assessing the importance of P-wave dispersion in dogs.

Methods

The current study was aimed at determining proper value of P_d in healthy dogs (group I), dogs with chronic valvular disease (group II) and dogs with disturbances of supraventricular conduction (group III). The tests were carried out in 53 healthy dogs, 23 dogs with chronic valvular disease and 12 dogs with disturbances of supraventricular conduction of various breeds, sexes and body weight from 1,5 to 80 kg, aged between 0,5 and 17 years, submitted to the ECG examination. ECG was acquired in dogs in a standing position with BTL SD-8 electrocardiographic device and analyzed once the recording was enlarged. P-wave duration was calculated in 9 ECG leads (I, II, III, aVR, aVL, aVF, V1, V2, V4) from 5 cardiac cycles.

Results

The proper P-wave dispersion in healthy dogs was determined at up to 24 ms. P-wave dispersion was statistically significant increased (p < 0.01) in dogs with chronic valvular disease and dogs with disturbances of supraventricular conduction. In dogs with the atrial enlargement the P-wave dispersion is also higher than in healthy dogs, although no significant correlation between the size of left atria and Pd was noticed (p = 0.1, r = 0,17).

Conclusions

The P-wave dispersion is a constant index in healthy dogs, that is why it can be used for evaluating P wave change in dogs with chronic valvular disease and in dogs with disturbances of supraventricular conduction.     

Premature centromere division (PCD) identified in a hucul mare with reproductive difficulties

A hucul mare with reproductive abnormalities was examined during karyotype analysis. The karyotype was analysed based on evaluation of 860 metaphase plates in chromosome preparations. The use of fluorescence in situ hybridization (FISH) with an X chromosome painting probe showed premature X chromosome separation in 9.5% cases of examined chromosome spreads. In this report, we present the first identify premature centromere division (PCD) as a possible cause of abnormal X chromosome morphology in horses and as a probable cause of reproductive difficulties.     

Characteristics of capercaillie (Tetrao urogallus) semen analysed with flow cytometry combined with fertility results

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.     

Analysis of introgression of the Tulipa fosteriana genome into Tulipa gesneriana using GISH and FISH

Southern hybridization and genomic in situ hybridization (GISH) have demonstrated that ‘Purissima’ (2n = 2x = 24) is an interspecific hybrid comprised of one genome of Tulipa (T.) gesneriana and one genome of T. fosteriana. Backcrossing T. gesneriana with ‘Purissima’ was partially successful. Simultaneous GISH and fluorescence in situ hybridization (FISH) distinguished chromosomes from both parent genomes, as well as recombinant chromosomes, in interspecific hybrids and their progeny. Chromosome recombination was observed in all cultivars except ‘Purissima’ and ‘Kouki’ (2n = 3x = 36). ‘Kouki’ (2n = 3x = 36) had two genomes of the T. gesneriana and a single genome of the T. fosteriana. The number of nonrecombinant T. fosteriana chromosomes in ‘Judith Leyster’ (2n = 4x = 48) and ‘Purissima’ progeny varied from two in ‘Hatsuzakura’ to six in ‘Kikomachi’ and ‘Momotaro’. The number and type of recombinant chromosomes also differed among cultivars. The total number of translocations ranged from one in ‘Kikomachi’ to six in ‘Hatsuzakura’. Each was a combination of a single T. fosteriana fragment and a single T. gesneriana fragment, indicating that they resulted from a single crossover event. Sequential GISH and FISH analysis with rDNA probes yielded chromosome-specific markers that were used to identify most of the chromosomes in ‘Purissima’ progeny. This is the first report of introgression of T. fosteriana chromatin into the T. gesneriana genome.     

Identification of S-haplotypes of European cultivars of sour cherry

The sour cherry is known to exhibit the phenomenon of gametophytic self-incompatibility which prevents self-fertilization. In sour cherry, besides self-incompatible cultivars, there often occur self-compatible cultivars. This is due to the occurrence of natural mutations of the S-RNase or SFB genes and, consequently, loss of functionality of S-alleles. Here we present the results of the identification of S-haplotypes of 21 cultivars of sour cherry from various regions of Europe. The analyses were performed using methods based on the amplification of intron I and intron II of the S-RNase gene and fragments specific to the individual alleles of the S-RNase or SFB genes. The tested cultivars were found to contain 15 S-haplotypes: S₁, S_1?, S₄, S₆, S_6m, S_6m2, S₉, S₁₂, S₁₃, S_13?, S₂₆, S₃₅, S_36a, S_36b, and S_36b2. The most frequently occurring S-haplotypes were S_13? (61.9%), S_36a (57.1%), and S₂₆ (47.6%). On the basis of the results, 17 of the 21 cultivars were deduced to be self-compatible. The results will be of use in the production of sour cherry fruit by facilitating the selection of suitable pollinating cultivars. The results are also expected to be useful in the breeding of new cultivars of sour cherry when selecting genotypes for crosses.     

Evaluation of the ability of colistin,amoxicillin (components of Potencil®), and fluoroquinolones to attenuate bacterial endotoxin‐ and Shiga exotoxin‐mediated cytotoxicity—In vitro studies

Escherichia coli is one of the major pathogens in humans and animals causing localized and systemic infections, which often lead to acute inflammation, watery diarrhea, and hemorrhagic colitis. Bacterial lipopolysaccharide (LPS) and Shiga exotoxins (Stx) are mostly responsible for such clinical signs. Therefore, highly effective treatment of E. coli infections should include both eradication of bacteria and neutralization of their toxins. Here, for the first time, we compared the in vitro ability of common antibiotics to decrease LPS‐ and Stx‐mediated cytotoxicity: colistin, amoxicillin (used separately or combined), enrofloxacin, and its metabolite ciprofloxacin. Three experimental scenarios were realized as follows: (a) the direct effect of antibiotics on endotoxin, (b) the effect of antibiotic treatment on LPS‐mediated cytotoxicity in an experiment mimicking “natural infection,” (c) the effect of antibiotics to decrease Stx2e‐mediated cytotoxicity. Two cell lines, A549 and Vero cells, were used to perform cytotoxic assays with the methyl tetrazolium (MTT) and lactate dehydrogenase leakage (LDH) methods, respectively. Colistin and amoxicillin, especially used in combination, were able to attenuate LPS toxic effect, which was reflected by increase in A549 cell viability. In comparison with other antibiotics, the combination of colistin and amoxicillin exhibited the highest boster or additive effect in protecting cells against LPS‐ and Stx2e‐induced toxicity. In summary, in comparison with fluoroquinolones, the combination of colistin and amoxicillin at concentrations similar to those achieved in plasma of treated animals exhibited the highest ability to attenuate LPS‐ and Stx2e‐mediated cytotoxicity.