Prevalence of trichomonads in the cloaca of wild wetland birds in the Netherlands.

Severe granulomatosis in productive layer chickens due to Tetratrichomonas gallinarum strain 13/16632 infection occurred in 2013 and 2017 on farms situated in a wetland area in the Netherlands. We hypothesized that wetland birds could be the source of the infection. Therefore, a prevalence study on trichomonads was performed by analysing cloaca swabs of 526 birds belonging to 13 species of wetland birds. The number of birds sampled ranged from 1 to 275 per species. Birds were sampled at 15 locations in the Netherlands. DNA extracted from the cloaca swabs was subjected to nested PCR using trichomonad-specific primers targeting the internal transcribed spacer 1 (ITS1)-5.8S rRNA-ITS2 region followed by cloning and sequencing. In nine bird species, trichomonads were detected; the overall prevalence was 9% (47/526), while the prevalence in the five species for which a substantial number of birds were examined (at least 39 per species) ranged from 4% to 24%. Three trichomonad species were found: T. gallinarum, Trichomonas tenax and Simplicimonas sp. of which T. gallinarum dominated. The virulent T. gallinarum strain 13/16632 was not detected, but closely related strains were. Phylogenetic analysis revealed that all T. gallinarum isolates belonged to two clusters within lineage 15 of Tetratrichomonas lineages. All T. tenax isolates were identical and clustered with reference strain H95, while Simplicimonas sp. isolates showed large genetic diversity. Some isolates may represent a new species of the genus Simplicimonas. We conclude that trichomonads are widespread amongst wetland birds, raising the question, amongst others, of their relevance for commercial poultry.RESEARCH HIGHLIGHTSTrichomonads occur among wild wetland birds in the Netherlands.T. gallinarum is the dominant trichomonad species in the cloaca of wetland birds.Some T. gallinarum isolates are closely related to a strain causing granulomas in layer chickens.Some isolates may represent a new species of the genus Simplicimonas.

Consequences of Keratin Phosphorylation for Cytoskeletal Organization and Epithelial Functions.

Intermediate filaments are major phosphoproteins. The complex patterns of intermediate filament phosphorylation make up a poorly understood code reflecting cytoskeletal properties and cellular function through an intense crosstalk with multiple signaling pathways. This review focuses on the epithelial keratin intermediate filaments highlighting the tight-knit relationship of keratin phosphorylation and network organization during cell division and apoptosis, and the importance of keratin phosphorylation during epithelial stress responses. The occurrence of keratin phosphorylation in genetic skin diseases and acquired diseases of simple epithelial tissues in liver, pancreas, and colon will be discussed. Finally, we will review the role of keratin phosphorylation in cancer with an emphasis on migration.

Production of interspecies chimeras by transplanting rosy barb (Puntius conchonius) embryonic cells to zebrafish (Danio rerio) embryos.

Establishment of a cell-mediated gene transfer system has potential as a new breeding technique for commercially valuable fishes. As an important step toward developing an inter-species chimera, cells from blastula-stage embryos of rosy barb (Puntius conchonius) were transplanted into zebrafish (Danio rerio) blastula-stage embryos to observe the development of the recipient. From the total of 473 transplants obtained only a fraction of 13 chimeras appeared perfectly normal after one month. Over two in normal 13 chimeras showed some characters from the donor cells with scarce pigmentation. This is the first successful inter-species study on zebrafish by using blastula cell transplants from rosy barb both belonging to the same family cyprinidae.

A clinical method for locating the femoral head centre during total knee arthroplasty.

Localisation of the femoral head centre (FHC) during total knee arthroplasty (TKA) is necessary when assessing the overall alignment of the leg using an extramedullary guide for the femur. On-table radiographs for this purpose are time consuming and inconvenient. We describe a clinical method for estimating the FHC: it is marked 1.5 cm lateral to the point where the femoral artery crosses the line joining the pubic tubercle and the anterior superior iliac spine. Using this method the estimated FHC was within 15 mm of the anatomic centre in 84% of cases, and within 24 mm in 99%. This translates into possible knee alignment errors of 2 degrees and 3 degrees, respectively. As alignment error of up to +/-3 degrees is compatible with satisfactory clinical results of TKA, our method provides useful approximation of the FHC. For greater accuracy however, an on-table radiograph is recommended.

Purification, characterization and antigenic species-specific reactivity of vitellogenin of rosy barb (Puntius conchonius Hamilton).

Vitellogenin (Vg) was isolated using gel filtration and ion-exchange chromatography from plasma of rosy barb (Puntius conchonius) treated with estrogen (estradiol-17beta). The purified Vg was stained positive for carbohydrate, lipid and phosphorus and was rich in Ala (10.58%), Asp (8.46%), Glu (10.30%), Leu (11.23%), Lys (7.22%) and Val (7.49%). It appeared as a single band of approximately 450 kDa in native PAGE and was reduced to a single band of approximately 167 kDa under SDS-PAGE, suggesting that it is probably composed of three identical polypeptide subunits. Double-immunodiffusion assay showed that the plasma from female rosy barb reacted with the mouse antisera against rosy barb Vg, forming a single immunoprecipitin line, while the plasma from male rosy barb or female zebrafish showed no such reactivity, confirming the existence of the sex- and species-specific reactivity for rosy barb Vg antisera.

Cryopreservation of flounder (Paralichthys olivaceus) sperm with a practical methodology.

A simple and convenient protocol for the cryopreservation of the flounder (Paralichthys olivaceus) sperm was established for "on the spot" cryopreservation of large quantities of semen. The use of three cryoprotectants, dimethyl sulphoxide (DMSO), glycerol (Gly) and methanol was tested in the method. The percentage of motile sperm present in semen after it had been frozen and thawed in the presence of DMSO, Gly or methanol was 60.5+/-3.6, 79.17+/-4.5 and 13.25+/-4.7%, respectively. The fertilization rates of this sperm were 67.06+/-15.1, 76.20+/-10.0 and 44.93+/-22.6%, while the hatching rates of eggs fertilized with this sperm were 37.40+/-8.3, 48.18+/-25.7 and 23.35+/-10.8%, respectively. It was found that Gly and DMSO were better cryoprotectants than methanol, with Gly giving the best overall results. Under scanning electron microscopy, it could be seen that while the majority of the frozen-thawed sperm remained morphologically normal, some exhibited lost or dilated mitochondria, swollen mid-pieces, broken tails, or damaged cell membrane, which probably caused the decrease in motility and fertility of the frozen-thawed sperm.

Study of solubility of isoproturon by its complexation with beta-cyclodextrin.

The inclusion complexes of beta-cyclodextrin-isoproturon were prepared using techniques such as kneading, co-evaporation, and co-precipitation. These complexes were characterised for physical properties and spectral studies such as UV, FTIR, and 1HNMR spectroscopy. To investigate the stoichiometry of the complexes Phase solubility method was adopted. Most of the complexes showed an increase in the dissolution rate of the herbicide. The best results were obtained when inclusion complexes were prepared by co-precipitation and co-evaporation method. Findings of this paper will allow the rational use of isoproturon with increase in its efficacy.

Pseudo-aneurysm of the anterior tibial artery complicating high tibial osteotomy--a case report.

The anterior recurrent branch of the anterior tibial artery is vulnerable during high tibial osteotomy. It may be injured during subperiosteal elevation of soft tissue from the lateral tibia--necessary for visualisation of the osteotomy or for placing the fixation device; or during excision of the infero-medial aspect of the fibular head--necessary to facilitate closure of the osteotomy. Recurrent bleeding from the operation site should draw attention to the possibility of a pseudo-aneurysm. Angiography helps in making an accurate diagnosis. Ligation of the feeding vessels provides effective treatment.