Artificial inoculation and susceptibility of Pinus armandii to Dothistroma septosporum

Dothistroma needle blight (DNB) is a serious needle disease of conifers that primarily affects pine species (Pinus spp.). Dothistroma septosporum is one of the DNB pathogens that has a diverse range of host species excluding Pinus armandii. In 15 inoculated P. armandii seedlings, D. septosporum acervuli were observed in 43 infected needles of ten seedlings with a mean disease severity of 1.11% at 25 weeks after inoculations, demonstrating the potential of D. septosporum to cause symptoms on the needles of P. armandii via artificial inoculation. The disease severity of P. armandii was similar to the positive control, Pinus nigra (median 0.75 for P. armandii to 0.70 for P. nigra), thus, P. armandii acts under artificial conditions as a susceptible host species.     

Specific contrast ultrasound using sterically stabilized microbubbles for early diagnosis of thromboembolic disease in a rabbit model

Specific contrast ultrasound is widely applied in diagnostic procedures on humans but remains underused in veterinary medicine. The objective of this study was to evaluate the use of microbubble-based contrast for rapid ultrasonographic diagnosis of thrombosis in small animals, using male New Zealand white rabbits (average weight about 3.5 kg) as a model. It was hypothesized that the use of microbubble-based contrast agents will result in a faster and more precise diagnosis in our model of thrombosis. A pro-coagulant environment had been previously established by combining endothelial denudation and external vessel wall damage. Visualization of thrombi was achieved by application of contrast microbubbles [sterically stabilized, phospholipid-based microbubbles filled with sulfur hexafluoride (SF₆) gas] and ultrasonography. As a result, rapid and clear diagnosis of thrombi in aorta abdominalis was achieved within 10 to 30 s (mean: 17.3 s) by applying microbubbles as an ultrasound contrast medium. In the control group, diagnosis was not possible or took 90 to 180 s. Therefore, sterically stabilized microbubbles were found to be a suitable contrast agent for the rapid diagnosis of thrombi in an experimental model in rabbits. This contrast agent could be of practical importance in small animal practice for rapid diagnosis of thrombosis.     

Deoxynivalenol and zearalenone residues in eggs of laying hens fed with a naturally contaminated diet: effects on egg production and estimation of transmission rates from feed to eggs

The potential for the Fusarium mycotoxins 4-deoxynivalenol (DON) and zearalenone (ZON) to enter the human food chain through contaminated eggs was assessed using a controlled feed study. Four groups of laying hens (eight in each group) were fed a diet that included differing amounts of naturally contaminated wheat containing DON ( approximately 20 mg kg(-1)) and ZON (0.5 mg kg(-1)). Eggs were collected and pooled from each group on a daily basis. Pooled samples were analyzed by liquid chromatography with mass spectrometry detection (LC-MS/MS). The method allowed DON, other type B trichothecenes, ZON, and its metabolites to be determined in a single multi-residue analysis. The selectivity of the MS/MS procedure allowed cleanup to be minimized (for DON, cleanup by immunoaffinity column was used) or eliminated (for ZON). The limits of detection of 0.01 microg kg(-1) for DON and 0.1 microg kg(-1) for ZON in eggs were lower than previously published methods. None of the samples analyzed had detectable levels of ZON or its metabolites. Although maximum levels of DON contamination (10 mg kg(-1) feed) were relatively high, no adverse effects were observed on egg production. On the basis of the determined DON levels in the hen's diet and the determined levels of DON in the corresponding eggs, transmission rates of 15 000:1, 18 000:1, and 29 000:1 for treatment levels 5, 7.5, and 10 mg DON kg(-1) feed, respectively, were found. These results show that, although eggs could be a human exposure route for DON, the levels are insignificant compared to the other sources, although the presence of metabolites of DON was not studied.     

Leukocytic responses and intestinal mucin dynamics of broilers protected with Enterococcus faecium EF55 and challenged with Salmonella Enteritidis

The protective effect of Enterococcus faecium EF55 in chickens challenged with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was assessed. The antibacterial effect on the bacterial microflora in the small intestine in relation to white blood cell count, phenotyping of peripheral blood and intestinal lymphocytes, functional activity of lymphocytes and phagocytes and mucin quantitation were investigated. Day-old chicks (85) were randomly divided into four groups. The probiotic group (EF) and Salmonella + probiotic group (EFSE) received E. faecium EF55 (10⁹ CFU – 3 g/group/day) for 21 days. The Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis (10⁸ CFU in 0.2 ml PBS) in a single dose per os on day four of the experiment. The control group chicks (C) were fed a commercial diet without added bacteria. Supplementation of EF55 in the diet of the chickens in the EFSE group, challenged with S. Enteritidis, caused the density of the intestinal mucin layer to increase significantly in non-specific regions (duodenum and jejunum), but decrease significantly in target regions (caeca) for S. Enteritidis. Probiotic treatment also appeared to result in a significantly higher number of lymphocytes in peripheral blood and a tendency to increase CD3, CD4, CD8, and IgM positive cells 3 days post-infection with S. Enteritidis. The results demonstrated an antibacterial effect and suggested that EF55 had a moderating effect on intestinal mucin production and leukocytic response in the early phase of S. Enteritidis infection.     

Effects of caprylic acid and triacylglycerols of both caprylic and capric acid in rabbits experimentally infected with enteropathogenic Escherichia coli O103

Eighty-eight rabbits weaned at the age of 35 days were divided into four groups. Rabbits of the first two groups were fed a granulated feed, free of antimicrobials. Rabbits of the 3rd and the 4th groups were fed the same diet, supplemented with caprylic acid at 5 g/kg, and with triacylglycerols (TAG) of caprylic and capric acid at 10 g/kg, respectively. Rabbits of the 2nd, 3rd and 4th groups were challenged orally with 10(9) cells of Escherichia coli of the O103 serogroup. Numbers of coliform bacteria in faeces of non-infected rabbits averaged 4.66 log(10)cfu/g. Six days after inoculation, caprylic acid and TAG of caprylic and capric acid decreased faecal output of coliforms from 10.18+/-0.62 to 7.79+/-0.48 log(10)cfu/g and 8.04+/-0.50 log(10)cfu/g, respectively. In the 2nd, 3rd and 4th groups 15, 11 and 9 infected rabbits died, respectively. However, the differences in mortality rate were not statistically significant. Surviving rabbits were slaughtered at 53 days of age. In caecal contents of infected rabbits, numbers of coliform bacteria were significantly reduced from 8.71 log(10)cfu/g to 5.55-5.83 log(10)cfu/g in treated groups. It can be concluded that both antimicrobial lipids are active against coliform bacteria, and may improve the resistance of weaned rabbits to enterocolitis.     

Needle blight caused by Dothistroma pini in Slovakia: distribution,host range and mating types

Dothistroma needle blight (DNB) has been observed in Slovakia during the last two decades. Up until 2017, Dothistroma septosporum has only been detected and molecularly confirmed to cause DNB in Slovakia. Here, we report the detection of Dothistroma pini at six localities around Slovakia, representing different plantation types. Four pine species (Pinus sylvestris, P. nigra, P. mugo and P. jeffreyi) were confirmed as hosts of D. pini in Slovakia, of which only P. mugo has been previously reported as host in Slovakia. Three gene regions (ITS, EF1 –α, and ß-tubulin) of each of the 13 isolates were sequenced and assigned as D. pini. Based on ITS sequences, the studied isolates represent the haplotypes Dp_HAP.1, Dp_HAP.2. Both mating types were detected but at different localities. Our results suggest that in addition to D. septosporum, D. pini may contribute to DNB also in Slovakia.     

The occurrence of reflexes and reactions in cattle following stunning with a captive bolt at the slaughterhouse

The difference in the frequency of the occurrence of reflexes/reactions in 355 cows (average weight 505.5 ± 7.9 kg) and 262 bulls (average weight 735.6 ± 8.4 kg) following stunning with a Matador SS 3,000 B trigger‐activated captive bolt gun (Termet) was determined. The stun shot more than 2 cm from the ideal position was found in 79.6% of animals. Vocalization, corneal reflex, rhythmic breathing, blinking, eyeball rotation and the absence of tongue protrusion occurred more frequently in bulls (p < .05). Spontaneous limb movements and nystagmus occurred more frequently in cows (p < .05). No difference between bulls and cows was determined in the occurrence of a response to painful stimuli or attempts to regain normal posture. No dependency was determined between the frequency of the occurrence of a reflex/reaction and the distance of the stun shot from the ideal point on the skull for any of the monitored signs. This study shows that the occurrence of reflexes/reactions following the stunning of cattle with a captive bolt is not only dependent on the position of the shot if placed within a 9 cm radius from the ideal point on the cattle skull.     

Drivers of forest fire occurrence in the cultural landscape of Central Europe

Context

Wildfires in temperate Central Europe have traditionally been perceived as a mere consequence of human activity without any relevance to natural forest development, despite their documented frequent occurrence. As a result, knowledge about local fire ecology and patterns of wildfire occurrence in the landscape is lacking.

Objectives

We aimed to reveal the factors influencing the spatial distribution of forest fires in the Czech Republic as a model area for the broader region. Specifically, we aimed to (1) find out which factors influence the occurrence and frequency of the forest fires at the country scale and in a selected fire-prone region; (2) examine the relationship of lightning strikes and their polarity with wildfire incidence; (3) identify the conditions determining areas with naturally driven fire-prone conditions.

Methods

We took data on 15,985 wildfire records and explored their spatial distribution using GIS layers of human, topographic, climatic and vegetation composition factors. We analysed the data using GLM and hierarchical partitioning methods.

Results

Wildfire occurrence was controlled mostly by environmental factors whereas wildfire frequency was strongly driven by human factors. In the selected fire-prone region, the effect of environmental factors was even more pronounced and wildfire frequency was also driven, albeit marginally, by lightning strikes of positive polarity.

Conclusion

The pattern of wildfire occurrence in the Czech Republic was similar also to those from regions where wildfire is considered a natural part of local ecosystems. We identified the areas with natural fire-prone conditions which probably led to the development of local fire-adapted ecosystems.

     

Immunohistochemical localization of phospholipase D in strawberry (Fragaria ananassa Duch.) fruits

Phospholipase D, the key enzyme involved in membrane phospholipid catabolism, was localized in strawberry (Fragaria ananassa Duch.) fruits during development using immunohistochemical techniques. Typical structural features such as the nucleus, cell wall, plasma membrane, mitochondria and vacuole were noticeable in cells of young strawberry fruits. Cytoplasmic structural features were less pronounced in fully grown and turning red fruits, except cell wall, which was considerably enlarged. Also, extensive microvesiculation from the plasma membrane was noticeable in ripening fruits. The number of visible gold particles indicative of phospholipase D increased during development. Phospholipase D was found to be associated with membranous structures and vacuole. Interestingly, gold-particles could also be observed in the cell wall space in ripening fruits suggesting that cytosolic contents may have leaked into the cell wall space as a result of membrane deterioration and loss of compartmentalization. The role of phospholipase D during development and ripening of strawberry, a non-climacteric fruit, is discussed.