Bone Marrow Transplantation Restores Follicular Maturation and Steroid Hormones Production in a Mouse Model for Primary Ovarian Failure

Recent studies suggest that bone marrow stem cells (BMSCs) are promising grafts to treat a variety of diseases, including reproductive dysfunction. Primary ovarian failure is characterized by amenorrhea and infertility in a normal karyotype female, with an elevated serum level of follicle-stimulating hormone (FSH) and a decrease level of estrogen caused by a mutation in FSH receptor (FSHR) gene. Currently, there is no effective treatment for this condition. The phenotype of FSHR (−/−) mouse, FORKO (follitropin receptor knockout), is a suitable model to study ovarian failure in humans. Female FORKO mice have elevated FSH, decreased estrogen levels, are sterile because of the absence of folliculogenesis, and display thin uteri and small nonfunctional ovaries. In this study, we determined the effects of BMSC transplantation on reproductive physiology in this animal model. Twenty four hours post BMSC transplantation, treated animals showed detectable estroidogeneic changes in daily vaginal smear. Significant increase in total body weight and reproductive organs was observed in treated animals. Hemotoxylin and eosin (H&E) evaluation of the ovaries demonstrated significant increase in both the maturation and the total number of the follicles in treated animals. The FSH dropped to 40–50% and estrogen increased 4–5.5 times in the serum of treated animals compared to controls. The FSHR mRNA was detected in the ovaries of treated animals. Our results show that intravenously injected BMSCs were able to reach the ovaries of FORKO mice, differentiate and express FHSR gene, make FSHR responsive to FSH, resume estrogen hormone production, and restore folliculogenesis

Simplified method to measure glucocorticoid metabolites in faeces of horses

Glucocorticoids or their metabolites can be measured in several body fluids or excreta, including plasma, saliva, urine and faeces. In recent years the measurement of glucocorticoid metabolites (GCMs) in faeces has gained increasing attention, because of its suitability for wild populations. In horses, however, the group-specific enzyme immunoassay described so far has a limited practicability due to its complex extraction procedure. Therefore, we tested the applicability of other enzyme immunoassays for glucocorticoid metabolites. The present study clearly proved that an enzyme immunoassay (EIA) for 11-oxoaetiocholanolone using 11-oxoaetiocholanolone-17-CMO: BSA (3alpha,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. Therefore it was possible to use just a small amount of the supernatant of a methanolic suspension of faeces. The results correlated well with the already described method for measuring GCMs in horse faeces, i.e. analysing the samples with an EIA after a two step clean up procedure of the samples (Merl et al. 2000). In addition, the 3alpha,11-oxo-A EIA has the advantage of providing a bigger difference between baseline values and peak values after ACTH stimulation. The new assay increased the accuracy of the test, lowered the expenses per sample, and storing samples at room temperature after collection was less critical than with other assays investigated in our study. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports and it shows the importance of choosing an assay which is in good accordance with the metabolites excreted in a given species

Molecular screening of pathogenic and symbiotic bacterial species in African ticks

INTRODUCTION. As haematophagous ectoparasites, hard ticks (Acari:Ixodidae) are vectors of numerous disease-causing bacterial pathogens worldwide. In Africa, the most important tick-borne bacteria are considered those belonging to the genera Rickettsia, Anaplasma, Ehrlichia, Babesia, and Theileria (Jongejan & Uilenberg, 2004, Parasitology, 129:S3-S14). On the other hand, very few data on bacterial symbionts of African ticks are found in the literature. Considering that symbiotic bacteria can play a key role in the host biology, and that they may also compete with or be beneficial towards pathogens transmission and biology, we investigated the co-presence of pathogenic and symbiotic bacteria in ticks collected in Africa. MATERIALS AND METHODS. Taxonomic identification of ticks was performed by means of both morphological (light microscopy) and molecular (PCR) techniques. The ticks were then screened for the presence of bacterial species through ad-hoc molecular approaches (PCR). RESULTS AND CONCLUSION. 250 ticks collected from Kenya, Nigeria, Madagascar, Egypt, and South Africa were taxonomically identified as belonging to Ixodes, Amblyomma, Hyalomma, Rhipicephalus, Dermacentor, and Haemaphysalis genera. Pathogen screening provides additional information on pathogens circulation in Africa, confirming the presence of Rickettsia spp., Anaplasma spp., Borrelia spp., Babesia spp. and Theileria spp. Furthermore, our work provides insights on the African scenario of tick-symbiont associations, laying the foundation for functional studies aimed at interaction analyses with possible implications on pest control