squashing method
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In this study, lactic acid bacteria (LAB) isolation from fermented foods and molecular identification using magnetic bead technology were performed. And then exopolysaccharide (EPS) production possibility was tested in agar medium, and the positive ones were selected for the next step. The bacteria that could produce higher carbohydrate level were grown in MRS medium fortified with whey and pumpkin waste. In our study, 19 different LAB species were identified from fermented products collected from different places in Hatay (Türkiye) province. In molecular identification, universal primer pairs, p806R/p8FPL, and PEU7/DG74 were used for PCR amplification. After that, PCR products purified using paramagnetic bead technology were sequenced by the Sanger sequencing method. The dominant species, 23.8% of the isolates, were identified as Lactiplantibacillus plantarum. As a technological property of LAB, exopolysaccharide production capability of forty-two LAB isolate was tested in agar medium, and after eleven isolates were selected as positive. Two LAB (Latilactobacillus curvatus SHA2-3B and Loigolactobacillus coryniformis SHA6-3B) had higher EPS production capability when they were grown in MRS broth fortified with pumpkin waste and whey. The highest EPS content (1750 mg/L glucose equivalent) was determined in Loigolactobacillus coryniformis SHA6-3B grown in MRS broth fortified with 10% pumpkin waste. Besides the produced EPS samples were validated with FTIR and SEM methods.
- Klíčová slova
- Exopolysaccharide, Magnetic bead, Pumpkin waste, Sanger sequencing, Whey,
- MeSH
- bakteriální polysacharidy * biosyntéza metabolismus MeSH
- Cucurbita mikrobiologie MeSH
- fermentace MeSH
- fermentované potraviny * mikrobiologie MeSH
- fylogeneze MeSH
- kultivační média chemie MeSH
- Lactobacillales * izolace a purifikace klasifikace genetika metabolismus MeSH
- odpadní produkty * analýza MeSH
- potravinářská mikrobiologie * MeSH
- RNA ribozomální 16S genetika MeSH
- syrovátka MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bakteriální polysacharidy * MeSH
- kultivační média MeSH
- odpadní produkty * MeSH
- RNA ribozomální 16S MeSH
This review paper focuses on the recent advancements in the large-scale and laboratory-scale isolation, modification, and characterization of novel starches from accessible botanical sources and food wastes. When creating a new starch product, one should consider the different physicochemical changes that may occur. These changes include the course of gelatinization, the formation of starch-lipids and starch-protein complexes, and the origin of resistant starch (RS). This paper informs about the properties of individual starches, including their chemical structure, the size and crystallinity of starch granules, their thermal and pasting properties, their swelling power, and their digestibility; in particular, small starch granules showed unique properties. They can be utilized as fat substitutes in frozen desserts or mayonnaises, in custard due to their smooth texture, in non-food applications in biodegradable plastics, or as adsorbents. The low onset temperature of gelatinization (detected by DSC in acorn starch) is associated with the costs of the industrial processes in terms of energy and time. Starch plays a crucial role in the food industry as a thickening agent. Starches obtained from ulluco, winter squash, bean, pumpkin, quinoa, and sweet potato demonstrate a high peak viscosity (PV), while waxy rice and ginger starches have a low PV. The other analytical methods in the paper include laser diffraction, X-ray diffraction, FTIR, Raman, and NMR spectroscopies. Native, "clean-label" starches from new sources could replace chemically modified starches due to their properties being similar to common commercially modified ones. Human populations, especially in developed countries, suffer from obesity and civilization diseases, a reduction in which would be possible with the help of low-digestible starches. Starch with a high RS content was discovered in gelatinized lily (>50%) and unripe plantains (>25%), while cooked lily starch retained low levels of rapidly digestible starch (20%). Starch from gorgon nut processed at high temperatures has a high proportion of slowly digestible starch. Therefore, one can include these types of starches in a nutritious diet. Interesting industrial materials based on non-traditional starches include biodegradable composites, edible films, and nanomaterials.
- Klíčová slova
- amylose–lipid complexes, edible films, granule size, nanomaterials, resistant starch, starch analyses, temperature of gelatinization,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
BACKGROUND: Trypanosoma theileri species complex includes parasites of Bovidae (cattle, sheep, goat, etc.) and Cervidae (deer) transmitted mainly by Tabanidae (horse flies and deerflies) and keds (Hippoboscidae). While morphological discrimination of species is challenging, two big clades, TthI and TthII, each containing parasites isolated from bovids and cervids, have been identified phylogenetically. To date, the development in the vector has been studied in detail only for the ked-transmitted sheep parasite T. melophagium (TthII), while the fate of trypanosomes in tabanids was described only briefly by light microscopy. METHODS: We collected infected tabanids of various species and identified trypanosomes by molecular phylogenetic analysis. The morphology and development of trypanosomes was studied using the combination of statistical analyses as well as light and electron microscopy. RESULTS: Two trypanosome species belonging to both TthI and TthII clades of the T. theileri complex were identified. The phylogenetic position of these two trypanosomes suggests that they parasitize deer. Both species were indiscernible by morphology in the vector and showed the same development in its intestine. In contrast to the previously described development of T. melophagium, both trypanosomes of tabanids only transiently infected midgut and settled mainly in the ileum, while pylorus and rectum were neglected. Meanwhile, the flagellates developing in the tabanid ileum (pyriform epimastigotes and metacyclic trypomastigotes) showed similarities to the corresponding stages in T. melophagium by morphology, mode of attachment to the host cuticle and formation of the fibrillar matrix surrounding the mass of developing parasites. In addition, for the first time to our knowledge we documented extraintestinal stages in these trypanosomes, located in the space between the epithelium and circular muscles. CONCLUSIONS: The development of different species of flagellates of the T. theileri complex in their insect vectors shows many similarities, which can be explained not only by their common origin, but also the same transmission mode, i.e. contamination of the oral mucosa with the gut content released after squashing the insect either by tongue or teeth. The observed differences (concerning primarily the distribution of developmental stages in the intestine) are associated rather with the identity of vectors than the phylogenetic position of parasites.
- Klíčová slova
- Deerflies, Horseflies, Life cycle, Trypanosomes, Vector,
- MeSH
- Diptera * parazitologie MeSH
- fylogeneze MeSH
- hmyz - vektory parazitologie MeSH
- ovce MeSH
- skot MeSH
- Trypanosoma * MeSH
- vysoká zvěř * parazitologie MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In the article we summarize the most common recent cytogenetic methods used in analysis of karyotypes in Heteroptera. We seek to show the pros and cons of the spreading method compared with the traditional squashing method. We discuss the suitability of gonad, midgut and embryo tissue in Cimex lectularius Linnaeus, 1758 chromosome research and production of figures of whole mitosis and meiosis, using the spreading method. The hotplate spreading technique has many advantages in comparison with the squashing technique. Chromosomal slides prepared from the testes tissue gave the best results, tissues of eggs and midgut epithelium are not suitable. Metaphase II is the only division phase in which sex chromosomes can be clearly distinguished. Chromosome number determination is easy during metaphase I and metaphase II. Spreading of gonad tissue is a suitable method for the cytogenetic analysis of holokinetic chromosomes of Cimex lectularius.
- Klíčová slova
- holokinetic chromosomes, karyogram, midgut, spreading method, squashing method, testes,
- Publikační typ
- časopisecké články MeSH
A new species of the family Alloionematidae was isolated from a rotten winged gourd at White Crane Garden, San Francisco, USA, sampled by Christopher Nelson in November 2010, and a live culture is deposited in Félix Lab Strain Database (http://www.justbio.com/worms/index.php), IBENS, Paris, France. Specimens from the culture have been examined. Both morphologically and molecularly, the nematode described herein as Alloionema californicum n. sp. differs from the other alloionematid species, A. appendiculatum and Neoalloionema tricaudatum. It is characterised by having a narrow stoma, 2.5-3.5 or 4 times longer than broad in adults or dauer juveniles respectively. Lateral fields are not present in adults but occur as one prominent ridge in dauers. Males have no bursa, six pairs of genital papillae and one single papilla. Dauers have large apparent phasmids in the middle of the tail. The ecology of the newly described species is unknown but probably it is a saprobic bacteriophagous nematode preferring rotting organic material.
- Klíčová slova
- Nematoda, Alloionema appendiculatum, Neoalloionema tricaudatum, ITS rDNA, SSU rDNA, phylogenetic analysis,
- MeSH
- Cucurbita parazitologie MeSH
- fylogeneze MeSH
- nemoci rostlin parazitologie MeSH
- Rhabditida klasifikace genetika izolace a purifikace ultrastruktura MeSH
- sekvenční analýza DNA MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Spojené státy americké MeSH
Migration of parasitic worms through the host tissues, which may occasionally result in fatal damage to the internal organs, represents one of the major risks associated with helminthoses. In order to track the parasites, traditionally used 2D imaging techniques such as histology or squash preparation do not always provide sufficient data to describe worm location/behavior in the host. On the other hand, 3D imaging methods are widely used in cell biology, medical radiology, osteology or cancer research, but their use in parasitological research is currently occasional. Thus, we aimed at the evaluation of suitability of selected 3D methods to monitor migration of the neuropathogenic avian schistosome Trichobilharzia regenti in extracted spinal cord of experimental vertebrate hosts. All investigated methods, two of them based on tracking of fluorescently stained larvae with or without previous chemical clearing of tissue and one based on X-ray micro-CT, exhibit certain limits for in vivo observation. Nevertheless, our study shows that the tested methods as ultramicroscopy (used for the first time in parasitology) and micro-CT represent promising tool for precise analyzing of parasite larvae in the CNS. Synthesis of these 3D imaging techniques can provide more comprehensive look at the course of infection, host immune response and pathology caused by migrating parasites within entire tissue samples, which would not be possible with traditional approaches.
- Klíčová slova
- 3D imaging, Light sheet microscopy, Neuroinfection, Parasite migration, Trichobilharzia regenti, X-ray micro-CT,
- MeSH
- infekce červy třídy Trematoda veterinární MeSH
- larva MeSH
- nemoci zvířat diagnóza parazitologie MeSH
- obratlovci MeSH
- protozoární infekce centrálního nervového systému veterinární MeSH
- Schistosomatidae * MeSH
- zobrazování trojrozměrné metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Despite remarkable recent progress in the analysis of plant genome organization and chromosome structure, there is a need for methods enabling DNA sequences to be mapped by fluorescence in situ hybridization (FISH) at high spatial resolution. We sorted mitotic metaphase chromosomes of wheat by flow cytometry and observed the occurrence of hyperexpanded chromosomes among them. However, this phenomenon was not reproducible in subsequent experiments. An investigation into the procedures of flow cytometry revealed that the hyperexpansion of chromosomes became reproducible when the concentration of formaldehyde used in sample fixation was reduced. We conducted FISH analysis with 45S rDNA, 5S rDNA and wheat centromeric repeat sequences as probes on flow-sorted chromosomes and also on chromosomes from squash preparations. We measured the length of chromosomes 1B and 6B, identified by FISH. On average, the hyperexpanded 1B and 6B chromosomes were 7.26 and 7.53 times longer, respectively, than the same chromosomes from the squash preparations. The most stretched 1B and 6B chromosomes both exceeded 100 micrometers.
In South Bohemia, Czech Republic, 178 shrews, including 98 common shrews, Sorex araneus L., 70 pygmy shrews, Sorex minutus L., and 10 lesser white-toothed shrews, Crocidura suaveolens (Pallas), were examined for Soricimyxum fegati Prunescu, Prunescu, Pucek et Lom, 2007 infections, using squash preparations of unfixed tissues, histological sections and molecular methods. The infection was found in 51 (52%) S. araneus, 14 (20%) S. minutus and 1 (10%) C. suaveolens. The records of the latter two species extend host range of S. fegati. Lesions associated with S. fegati infections in the liver, the organ of specific localisation of the parasite, were found to be induced by proliferative stages migrating toward lumina of bile ducts. In other organs of these three host species, xenoma-like formations (XLFs) were found that severely injured blood vessels. XLFs contained presporogonic stages of S. fegati, whose species identity was evidenced using molecular methods.
- MeSH
- cévy parazitologie patologie MeSH
- druhová specificita MeSH
- hostitelská specificita * MeSH
- játra parazitologie patologie MeSH
- molekulární sekvence - údaje MeSH
- Myxozoa klasifikace genetika izolace a purifikace patogenita MeSH
- orgánová specificita MeSH
- parazitární nemoci u zvířat parazitologie patologie MeSH
- rejskovití klasifikace parazitologie MeSH
- RNA ribozomální analýza MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- žlučové cesty parazitologie patologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- RNA ribozomální MeSH
BACKGROUND AND AIMS: Despite extensive study of polyploidy, its origin, and ecogeographical differences between polyploids and their diploid progenitors, few studies have addressed ploidy-level structure and patterns of ecogeographical differentiation at various spatial scales using detailed sampling procedures. The pattern of coexistence of polyploids in the geophyte Allium oleraceum at the landscape and locality scale and their ecology were studied. METHODS: Flow cytometry and root-tip squashes were used to identify the ploidy level of 4347 plants from 325 populations sampled from the Czech Republic using a stratified random sampling procedure. Ecological differentiation among ploidy levels was tested by comparing sets of environmental variables recorded at each locality. KEY RESULTS: Across the entire sampling area, pentaploids (2n = 5x = 40) predominated, while hexaploids (2n = 6x = 48) and tetraploids (2n = 4x = 32) were less frequent. The distribution of tetra- and hexaploids was partially sympatric (in the eastern part) to parapatric (in the western part of the Czech Republic) whereas pentaploids were sympatric with other cytotypes. Plants of different ploidy levels were found to be ecologically differentiated and the ruderal character of cytotypes increased in the direction 4x --> 5x --> 6x with the largest realized niche differences between tetra- and hexaploids. Most populations contained only one ploidy level (77 %), 22 % had two (all possible combinations) and 1 % were composed of three ploidy levels. The majority of 4x + 5x and 5x + 6x mixed populations occurred in sympatry with uniform populations of the participating cytotypes in sites with ecologically heterogeneous or marginal environment, suggesting secondary contact between cytotypes. Some mixed 4x + 6x populations dominated by tetraploids being sympatric and intermixed with uniform 4x populations might represent primary zones of cytotype contact. Almost no mixed accessions were observed on the fine spatial scale in mixed populations. CONCLUSIONS: The results provide evidence for adaptive differences among ploidy levels, which may contribute to their complex distribution pattern. The prevalence of asexual reproduction, limited dispersal and equilibrium-disrupting processes may support local coexistence of cytotypes.
- MeSH
- Allium klasifikace genetika růst a vývoj MeSH
- ekologie * MeSH
- ploidie * MeSH
- polyploidie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
PURPOSE OF THE STUDY: To report on the surgical treatment of intra-articular calcaneal fractures by open reduction and internal fixation with a calcaneal locking compression plate (LCP) from an extended lateral approach, and to retrospectively analyze the mid-term results in a group of patients treated by this technique. MATERIAL: In the period from August 2005 till March 2007, a total of 49 patients with 61 calcaneal fractures were treated. Of these, 11 (18 %) were treated conservatively. Reduction combined with Kirschner-wire fixation was used in four fractures (6.6 %). Open reduction with internal calcaneal LCP fixation (ORIF- calcaneal LCP) from an extended lateral approach was carried out to treat 46 fractures (75.4 %) in 38 patients. The group evaluated here comprised 29 patients with 33 calcaneal fractures treated by ORIF-calcaneal LCP at a follow-up longer than 6 months. The fractures were classified on the basis of computer tomography (CT) findings as Sanders types I to IV. The group had two woman (6.9 %) and 27 men (93.1 %) with an average age of 34.2 years (range, 19-55 years). In 11 fractures (33.3 %), the primary treatment included filling a central cancellous bone defect area. Calcium phosphate bone substitute material (resorbable ChronOS) was used in nine cases (27.3 %), a self-solidifying hydroxyapatite implant was injected in two (6.1 %) cases (X3 Wright and Norian SRS, respectively), and a bone allograft was implanted in one case (3 %). METHODS: Indicated for surgery were patients with an intra-articular calcaneal fracture, Sanders type II or type III, with articular surface displacement by more than 1 mm. Contraindications included age over sixty years, poor cooperation, smoking habits, peripheral vascular disease or skin infection. Surgery was performed only after oedema had resolved. The aim of our treatment was to achieve anatomical reconstruction of all articular surfaces, to restore the height, length, width and axis of the heel bone, to carry out primary stable osteosynthesis, and to enable the patient to begin rehabilitation with early mobilization. A passive rehabilitation usually started on the first post-operative day, and full weight-bearing of the extremity was allowed not earlier than 3 months post-operatively. Regular check-ups were at 6 weeks, 3, 6 and 12 months and then every year. The mid-term results were evaluated by the system of Rowe et al., scoring rest pain, possibility of return to preinjury jobs, use of walking aids, restriction of physical activity and limping. RESULTS The most frequent cause of injury was a fall or jump from height; this was recorded in 27 patients (93.1 %). An open fracture was diagnosed on two occasions (6.9 %). Bilateral calcaneal fractures were found in six patients (20.7 %); four (13.8 %) were treated by bilateral ORIF-calcaneal LPC and two (6.9 %) underwent closed reduction on one and Kirschnerwire transfixation on the other extremity. A combined injury to the musculoskeletal system was diagnosed in 11 patients (38 %), in whom four (13.8 %) had a tibial pylon fracture of the contralateral limb and four (13.8 %) had a thoracolumbar spine fracture. The surgical procedure was performed on average within 11.7 days of injury, and the average hospital stay was 18.2 days (range, 6 to 18 days). Early post-operative complications were recorded in six patients (20.7 %). Wound dehiscence was found in two (6.9 %), necrosis of wound edges in two (6.9 %), and early superficial infection responding to antibiotic therapy also in two patients (6.9 %). Deep infection, non-union or post-operative compartment syndrome were not recorded. Excellent Rowe scores were achieved in 10 patients (34.5 %), good in 15 (51.7 %) and satisfactory in two (6.9 %). Only two patients (6.9 %) reported poor outcome. DISCUSSION: The methods of classification and treatment of calcaneal fractures continue to be a frequently discussed topic. The technique of ORIF-calcaneal LCP from an extended lateral approach has recently been preferred for patients with displaced Sanders type II or III calcaneal fractures. In agreement with other literature references, this approach allowed us to observe the fracture, to reduce both the subtalar and calcaneocuboid articulations, to stabilize the fracture by internal fixation and to begin early rehabilitation. Because of the risk of early complications, the timing of surgery and a thorough consideration of indications and contraindications are of principal importance.We agree with other authors that filling calcaneal bone defects is not necessary. Pre- and post-operative CT scans are necessary. In accordance with literature data, fractures developing compartment syndrome are indicated for urgent fasciotomy and ORIF-calcaneal LCP should be postponed. In patients with multiple trauma and also in those with open calcaneal fractures, a temporary stabilization with an external fixator medially is performed first, and then converted to a second-stage, ORIF-LCP procedure. CONCLUSIONS: The surgical treatment of displaced intra-articular fractures that involves open reduction from an extended lateral approach and internal fixation with a calcaneal LCP shows very good preliminary results. A CT examination is required for the diagnosis and classification of fractures and a correct indication for surgery. Good timing is of principal importance. An urgent surgical intervention is necessary in open fractures or in those in which soft tissues are squashed by bone fragments. In the other fractures, surgery is carried out after oedema subsidence. Foot compartment syndrome is a serious complication of calcaneal fractures and urgent fasciotomy is the only adequate therapy. Full weight-bearing of the operated extremity depends on the rate of bone healing; it is usually allowed at 3 months after surgery.
- MeSH
- dospělí MeSH
- fraktury kostí chirurgie MeSH
- kostní destičky * MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- patní kost diagnostické zobrazování zranění chirurgie MeSH
- radiografie MeSH
- vnitřní fixace fraktury metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH