Two bacterial strains, Am4 and G10 were isolated from rumen fluid of different ruminants: cow (Holstein-Friesian) and sheep (Slovenskè merino), respectively. They were isolated from different hosts and regions, but showed 99.2% similarity of the 16S rRNA genes. Both strains are versatile and ferment various sugars to mainly succinate and lactate and small amounts of acetate and formate. The 16S rRNA sequences of Am4 and G10 revealed that they belonged to the genus Actinomyces, and are related to Actinomyces ruminicola JCM 13352 with 97.0% and 97.4% similarity, respectively. DDH showed strain Am4 and G10 had only 55.8 and 43.3% similarity with the Actinomyces ruminicola JCM 13352, and had 69.9% similarity among each other. Comparing strain Am4 and G10, gANI value and dDDH were 92.9% and 68.6%, respectively. Additionally, AAI between the strains was 95.8%. MLSA of housekeeping genes showed difference of metG and pheS. The G+C% contents of strain Am4 and G10 were 69.8% and 68.5%, respectively. MK-10(H) was the principal quinone for strain Am4 (82%) and G10 (91%) with small amounts of MK-10(H) and MK-10(H) for both strains. Only MK-9(H) was detected in strain Am4. MALDI-TOF analysis of protein profiles also revealed that Am4 and G10 are different from each other and from Actinomyces ruminicola JCM 13352. Based on phylogenetic and physiological characteristics, together with genome comparison and MLSA we propose two novel species in the genus Actinomyces: Actinomyces succiniciruminis sp. nov. (type strain Am4=TISTR 2317=DSM 10376) and Actinomyces glycerinitolerans sp. nov. (type strain G10=TISTR 2318=DSM 10377).
Ivanová H., Hrehová Ľ., Pristaš P. (2016): First confirmed report on Fusarium sporotrichioides on Pinus ponderosa var. jeffreyi in Slovakia. Plant Protect. Sci., 52: 250-253.During 2014-2015, samples from ten 20-25 years old pine (Pinus ponderosa var. jeffreyi) trees showing wilt symptoms were collected from the Arborétum Mlyňany park. A disease was observed on 20% of the trees. The first symptoms are wilting, stunting, chlorosis, and discolouration of needles, which turned yellow on affected twigs, then red and finally they fell off. Isolations of the pathogen were done from the discoloured tissues of needles (twenty samples from each tree) on Potato Dextrose Agar. Colonies of the fungus (3-4 Petri dishes from each tree) were initially aerial, white or slightly violet, but with age they became red and red pigments were produced in agar. The observed micromorphological characteristics of the fungus, such as presence of simple and proliferating conidiophores with polyphialides, microconidia, macroconidial shape, and chlamydospore presence matched the description of Fusarium sporotrichioides. The identity of the fungus was confirmed by phylogenetic analysis of internal transcribed spacer (ITS) sequences. Sequence comparisons placed the fungus to the species F. sporotrichioides with similarity of 99.6% at the ITS sequence level.
Lactic acid bacteria are symbiotic bacteria that naturally reside in the gastrointestinal tract of honey bees. They serve a multitude of functions and are considered beneficial and completely harmless. In our experiments Lactobacillus plantarum strain B35, isolated from honey bee digestive tract, was modified using pAD43-25 plasmid carrying a functional GFP gene sequence (gfpmut3a) and used as a model for monitoring and optimisation of the mode of application. The establishment of this strain in honey bee digestive tract was monitored using GFP fluorescence. Three different modes of oral application of this strain were tested: water suspension of lyophilised bacteria, aerosol application of these bacteria and consumption of sugar honey paste containing the lyophilised lactobacilli. Two days after administration the L. plantarum B35-gfp was present throughout the honey bee digestive tract with 10-10 cfu/bee with highest count observed for aerosol application.
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