Morphometric and biochemical screening of old mulberry trees (Morus alba L.) in the former sericulture region of Slovenia

Over centuries, in many European countries, the white mulberry trees (Morus alba L.) became an integral part of the cultural landscape, bearing witness to past sericulture activities. The distribution records of white mulberry trees in the cultural landscape are incomplete and in general poorly documented. The aim of the presented research was to collect data regarding geographical locations of mulberry trees and to define their morphological and biochemical variability in Goriška region, one of the historical sericulture regions in Slovenia. Principal component analysis of all morphometrical leaf traits allowed us to characterize two separate groups of morphotypes. Recordings of the tree pruning management revealed that annual base cutting is traditionally used in Goriška region. Significant correlations between pruning management and leaf morphology traits showed that frequently pruned trees form larger leaves. Biochemically, mulberry leaves are shown to be rich in proteins containing threonine, arginine, asparagine, serine, and glutamine as the most prominent free amino acids. The main phenolic compounds were identified as caffeoylquinic acid derivatives, quercetin malonyl-hexoside, rutin, kaempferol acetyl-hexoside, quercetin-3-glucoside, and p-coumaric acid derivatives. The difference in concentrations of the investigated metabolites is correlated either with the pruning management or the morphotype. Pruning significantly affected the levels of asparagine, alanine, and serine, which were higher in the annually pruned trees regardless of the morphotype. Furthermore, we were able to confirm a significant effect of pruning on total phenolics as well as on the levels of rutin, quercetin malonyl-hexoside, and quercetin-3-glucoside contents. Multivariate analysis allowed us to determine seven chemotypes with distinctive biochemical traits. Our results are the basis for defining superior high-yielding genotypes with optimum metabolic composition for both silkworm feeding as well as for innovative usage in food processing and pharmaceutical industries

Spremenljivost vsebnosti skupnih proteinov, fenolov in tiolov, v listih starih genotipov murv v Goriški regiji

Preserving the plant genetic resources of genus Morus is insuff­cient but undoubtedly vital for conservation of the world’s germplasm for our successors. This research was focused on old mulberry varieties from the Gorizia region in Slovenia which were assessed for their contents on crucial metabolites (proteins, phenolics and thiols) in leaves regarding their antioxidant and nutraceutical potentials. Total proteins were measured spectrophotometrically by following the procedure of Bradford, the total phenolic contents were determined using the Folin-Ciocalteu method and thiols were established with monobromobimane fluorescent dye. The presented metabolite screening showed that some of the evaluated genotypes had higher concentrations of glutathione and were superior in contents of proteins and phenolics when compared to the results of other authors and could be propagated as highly recommendable feed for silkworms, and other animals.Ohranjanje genskih virov rodu Morus je nezadostno, čeprav je nedvomno bistvenega pomena za ohranjanje svetovne zapuščine za naše naslednike. Ta raziskava temelji na določitvi ključnih metabolitov (proteinov, fenolov in tiolov) v listih starih genotipovmurv iz Goriške regije v Sloveniji z namenom proučitve antioksidativne aktivnosti in hranilne vrednosti. Skupni proteini in fenoli so bili določeni spektrofotometrično s pomočjo Bradfordove in Folin-Ciocalteu metode, tioli so bili določeni s pomočjo HPLC po predhodnem markiranju s fluorescenčnim barvilom monobromobimane. Rezultati določitve metabolitov kažejo, da imajo nekateri genotipi visoko vsebnost glutationa,v primerjavi z rezultati drugih avtorjev so superiorni v vsebnosti proteinov in fenolov in bodov nadaljnjih raziskavah razmnoženi z namenom uporabe listov za krmo sviloprejk in ostalih živali

Mycoplasma gallisepticum and Mycoplasma synoviae express a cysteine protease CysP, which can cleave chicken IgG into Fab and Fc

Major poultry pathogens M. gallisepticum and M. synoviae share a gene encoding a putative cysteine protease CysP similar to papain cysteine (CaA subfamily). Comparison of the cysP gene sequences of 18 M. synoviae and 10 M. gallisepticum strains sequenced in this study showed polymorphisms, including deletions. Seven M. synoviae strains, including the type strain WVU 1853, had a 39 bp deletion in the 3\u27 end of the cysP gene. In the same cysP region, all M. gallisepticum strains showed a deletion of 66bp. Immunoblot analysis with specific antibpodies demonstrated that M. synoviae strains expressed CysP, which was approximately 65 kDa. both M. synoviae and M. gallisepticum were able to digest chicken IgG (cIgG). Incubation of cIgG (-170 kDa) with M. synoviae or M. gallisepticum cells (-15 h at 37oC) resultes in a papain-like cleavage pattern of cIgG and fragments corresponding to the antigenbinding fragment of IgG (Fab, -45kDa) and the crystallizable region fragment (Fc) of the IgG heavy chain (dimer of -60 kDa). Iodoacetamide (50 mM) prevented cleavage of cIgG by both Mycoplasma species. Following site-directed mutagenesis (eight TGA codons were changed to TGG) the cysP gene of M. synoviae ULB 925 was expressed as a His-taggede protein in a cell-free system. Purified recombinant CysP (rCysp-67kDa, pI-8) cleaved cIgG into Fab and Fc fragments. This indicates that CysP is responsible for the cIgG cleavage caused by M. synoviae and probbaly, by M. gallisepticum. This is the first evidence to our knowledge that mycopasmas have enzymes that can cleave the host IgG and indicates a novel strategy used by M. gallisepticum and M. synoviae for prolomged survival despite the antibody response of their host

Demonstration of neuraminidase activity in Mycoplasma neurolyticum and of neuraminidase proteins in three canine Mycoplasma species

Neuraminidases are virulence factors in many pathogenic microarganisms. They are present also in some Mycoplasma species that cause disease in birds, dogs and alligators. Thirty-seven Mycoplasma species have been examined previously for neuraminidase (sialidase) activity, whereas many of the species causing disease in man, ruminants, pigs, rodent and other animals have not. In this study neuraminidase enzymatic activity (NEAC) was examined in 45 previously untested Mycoplasma species, including those causing diseases in man, farm animals and laboratory animals. The only species in which NEAC was found was Mycoplasma neurolyticum, specifically, its type strain (Type A[up]T) which is capable of inducing neurologic signs in inoculated young mice and rats. The NEAC of washed cells was relatively weak, but it differed even more than 10-fold among cells of cultures derived from individual colonies of M. neurolyticum. A weak NEAC was also detected in the supernatant of the M. neurolyticum broth culture. Canine Mycoplasma spp. with high sialidase activity reported previously, Mycoplasma canis, Mycoplasma cynos and Mycoplasma molare had 100-fold more NEAC than M. neurolyticum, but apparent differences in NEAC levels existed among strains of M. canis and of M. cynos. Zymograms using neuraminidase-specific chromogenic substrate were used to showproteins havingNEAC. In M. canis (a field isolate Larissa and the type strain PG14[up]), M. cynos (isolate 896) and M. molare (type strain H542[up]) proteins with NEAC had molecular mases of - 130 kDA, 105 kDa and 110 kDa, respectively. Identification of these neraminidases could provide the basis for their molecular characterization

The Effect of Feeding with Central European Local Mulberry Genotypes on the Development and Health Status of Silkworms and Quality Parameters of Raw Silk

SIMPLE SUMMARY: Several regions of Slovenia and Hungary retained numerous centuries-old white mulberry trees, evidence of past sericultural activities, being traditionally used to feed the silkworm larvae. Attempts for the reintroduction of sericulture in these countries are ongoing. The current study assessed the suitability of the locally adapted mulberry trees for contemporary sericultural needs. Silkworm hybrids were fed with leaves of the selected local mulberry genotypes and the larvae performance parameters (bodyweight, spinning success, cocoon quantity, and quality) were compared to those fed with reference mulberry varieties. The chemical contents and nutritive parameters of the mulberry leaves were determined, and connections were predicted between selected leaf compounds and silkworm performance parameters. The local mulberries had higher total protein contents, and lower total phenolic contents and differed in some individual phenolics, macro- and microelements compared to the reference sericultural and fruit varieties. A combined positive influence of proteins, specific phenolics, and microelements on larval growth and silk thread parameters was predicted. The health status and gut microbiome compositions of larvae were also analyzed. The results of the study indicate that selected local Slovenian and Hungarian mulberry varieties are suitable for high-quality silk cocoon and raw silk production. ABSTRACT: Silkworm rearing activities ceased in the 1970′s in several European countries. Attempts on the re-establishment of ecological and sustainable sericulture in Slovenia and Hungary are ongoing. The aim of the study was to assess the usability of locally adapted mulberry genotypes for sericulture and to estimate connections between leaf compound and silkworm performance parameters. A controlled feeding experiment of silkworms was performed to test the influence of leaves from selected trees on the growth of larvae, the health and microbiological status of larvae (e.g., gut bacterial microbiome, Bombyx mori nucleopolyhedrovirus infection), weight of cocoons and raw silk parameters. The Slovenian and Hungarian mulberry genotypes had significantly higher total protein contents, and lower total phenolic contents and differed significantly in some individual phenolics compared to the reference sericultural and fruit varieties. Significant differences were found in the contents of the macro- and microelements, namely S, Mn, Fe, and Sr. Based on correlative statistics and multivariate analysis, a combined positive influence of proteins, specific phenolics, and microelements on larval growth and silk thread parameters was predicted. The results of the study indicate that selected local Slovenian and Hungarian mulberry varieties are suitable for high-quality silk cocoon and raw silk production