A successful protocol for achieving anhydrobiosis of Gallus Gallus Domesticus spermatozoa while maintaining their fertility IN VIVO.

Lyophilization of avian semen is a new method for gene pool preservation. The goal of this study was to develop a protocol for the lyophilization of rooster semen with preserved fertility. Red Rhode Island rooster ejaculates (n = 20) were assessed by volume, motility, and concentration of spermatozoa. They were pooled and diluted 1:1 with a medium LCM-T20 containing trehalose (9.5 mM), exposed at 5 °C for 40 min and centrifuged, and then the supernatant was removed. Media LCM-T with trehalose (1.75 M) was added and exposed for 10 min. The semen was frozen in a thin layer in glass vials. Samples were lyophilized for 2 h at -150 … -50 °C. The water content of the samples after lyophilization was 6.1 ± 0.5% (CV 20%). The sample was rehydrated with a medium LCM-GA5 containing hyaluronic acid (40mg/100 mL media). The total motility of the spermatozoa was 1.0 ± 0.3%. From artificial insemination of virgin hens (n = 12) with rehydrated semen, one fertilized egg was obtained from eight laid eggs. All samples of perivitelline membranes of the obtained eggs had points of interaction with the spermatozoa (7-37 pcs/cm2), which confirmed the presence of viable rehydrated spermatozoa in the genital tract of the hen. To create a dry biobank for poultry, the first protocol for lyophilization of rooster semen was developed to ensure sperm fertility in vivo.

Magnetically Controlled Carbonate Nanocomposite with Ciprofloxacin for Biofilm Eradication.

Biofilms are the reason for a vast majority of chronic inflammation cases and most acute inflammation. The treatment of biofilms still is a complicated task due to the low efficiency of drug delivery and high resistivity of the involved bacteria to harmful factors. Here we describe a magnetically controlled nanocomposite with a stimuli-responsive release profile based on calcium carbonate and magnetite with an encapsulated antibiotic (ciprofloxacin) that can be used to solve this problem. The material magnetic properties allowed targeted delivery, accumulation, and penetration of the composite in the biofilm, as well as the rapid triggered release of the entrapped antibiotic. Under the influence of an RF magnetic field with a frequency of 210 kHz, the composite underwent a phase transition from vaterite into calcite and promoted the release of ciprofloxacin. The effectiveness of the composite was tested against formed biofilms of E. coli and S. aureus and showed a 71% reduction in E. coli biofilm biomass and an 85% reduction in S. aureus biofilms. The efficiency of the composite with entrapped ciprofloxacin was higher than for the free antibiotic in the same concentration, up to 72%. The developed composite is a promising material for the treatment of biofilm-associated inflammations.

Phylogenetic position of the pelobiont Mastigamoeba aspera and revision of the family Mastigamoebidae (Archamoebae, Pelobiontida).

In this study, we presented the results of our molecular phylogenetic analysis of Archamoebae using a newly obtained sequence of the 18S rRNA gene of Mastigamoeba aspera, the type species of the genus. In addition, we characterized the general organization of the tubulin cytoskeleton of M. aspera by immunofluorescent microscopy and TEM. Our findings allowed us to revise the family Mastigamoebidae and establish within it two subfamilies: Mastigamoebinae for the genus Mastigamoeba and Seraviniinae subfam. nov. for the genera Seravinia gen. nov., Paramastigamoeba gen. nov., Iodamoeba and Endolimax. The type genera Mastigamoeba and Seravinia are clearly distinguished by the structure of the basal apparatus of flagella. In addition, the tubulin cytoskeleton of several Mastigamoeba spp. contains a rim of microtubules around the nuclei.

Intestinal ciliates (Ciliophora) from the wild plains zebra (Equus quagga) in South Africa, with notes on the microtubule cytoskeleton organisation.

Investigation of the ciliate communities from the digestive tract of different wild vertebrates is important in context of host-specificity of different ciliate species and the detection of any cases of non-specific infection. Here we present a description and analysis of the fauna of ciliates (Litostomatea, Trichostomatia) inhabiting the intestine of the wild plains zebra (Equus quagga Boddaert, 1785) in South Africa. Nineteen species belonging to 12 genera of five families were found. Five species were specific to Equus quagga; one was also found in Equus zebra; 29 are common to different equids; and one had been previously described from rhinoceros. For the first time, we used immunofluorescent staining to investigate microtubule cytoskeletons in trichostomatids. We found that this staining method is useful for the identification of trichostomatids.

The nature and features of organization of reserve polysaccharides in three Pelomyxa species (Archamoebea, Pelobiontida).

The nature and features of organization of reserve polysaccharides in three species of the genus Pelomyxa-P. palustris, P. belevskii, and P. stagnalis-were studied using light and transmission electron microscopy. We applied the periodic acid-Schiff reaction that is a highly selective method for detecting glycogen. The fluorescent dye auramine-SO2 (Au-SO2) was used as a Schiff-type reagent. The densely packed aggregates of glycogen that form the morphologically differentiated organelle-like bodies are revealed in the cytoplasm in all studied species. The organization of these bodies is characterized by the species-specific features, while in most cases, their size and number in the cells vary depending on the season of the year. Although in all the cases we studied, these bodies do not have their own boundary membrane, in fact, they are surrounded by membranous structures. These structures differ in a variety of Pelomyxa species. We concluded that there are two groups of species in the genus Pelomyxa. The first one includes organisms containing glycogen structures in the cytoplasm (P. palustris, P. belevskii, P. stagnalis, P. binucleata, P. corona, P. secunda). No inclusions resembling glycogen bodies were found in P. flava, P. paradoxa, P. gruberi, and P. prima that form the second group.

Thin and Flexible Ion Sensors Based on Polyelectrolyte Multilayers Assembled onto the Carbon Adhesive Tape.

A novel flexible ion-selective sensor for potassium and sodium detection was proposed. Flexible ion-selective electrodes with pseudo-liquid internal solution on contrary to the system with a solid contact provided a more stable analytical signal. Such advantages were achieved because of polyelectrolyte (PEI/PSS) layers adsorption on the conduct substrate with a layer-by-layer technique. Such an approach demonstrated that ion-selective electrodes save sensitivity with Nernstian dependence: 56.2 ± 1.4 mV/dec a Na+ and 56.3 ± 1.9 mV/dec a K+ , as well as a fast time of response for potassium (5 s) and sodium (8 s) was shown. The sensing platform proposed demonstrates a better time of response and is close to the Nernstian value of sensitivity with a sensor low cost. The results proposed confirm a pseudo-liquid junction for the ion-selective electrode. Biocompatibility of an ion-selective sensing platform was demonstrated at potassium potentiometric measurements in Escherichia coli biofilms. Potassium levels in a biofilm were measured with potentiometry and showed agreement with the previous results.