Two successful embryo transfers of mini-donkey embryos in Brazilian Northeastern jennies using an alternative method: Case report     

Lucas T. Rodrigues  Lorenzo GTM Segabinazzi  Sidnei N Oliveira  Mariana Frasson  Frederico O Papa 《Reproduction in domestic animals》2021,56(11):1470-1474

Pregnancy rates after embryo transfer (ET) are disappointing in donkey species. This study aims to report two successful ET of mini-donkey embryos using Brazilian Northeastern jennies as recipients. Eighteen embryo flushes were performed 9 days post-ovulation in two non-pregnant mini-donkeys jennies (11 and 7 cycles per jenny). Eleven embryos (61%, 11/18) were collected and transferred to Brazilian Northeastern jennies 4–6 days post-ovulation by conventional (n = 6) or an alternative (n = 5) technique. The alternative method consisted of inserting a Polansky equine vaginal speculum smeared with lubricant in the vagina of the recipient jenny. The arms of the speculum were extended to allow the visualization of the cervix. Then, using an adapted crafted, elongated, toothed tissue grasping forceps, the external cervical os was held, and the cervix was gently pulled backward, aiming to straight the cervical canal. The ET gun was inserted through the vagina and cervix by visual inspection, and the embryo was released into the uterine lumen. All embryos collected were Grade 1 and classified as Expanded Blastocysts. No jennies become pregnant after conventional ET (0/6), whereas two recipient jennies (40%, 2/5) become pregnant and delivered offspring in the following year after ET using the alternative technique. In conclusion, Brazilian Northeastern jennies can be used as embryo recipients using the alternative method proposed in the present study. However, further investigations are needed to improve the knowledge and results of ET in donkey species.  相似文献   

An improved protocol for the production of AFLP^TM markers in complex genomes by means of capillary electrophoresis     

R. Papa  M. Troggio  P. Ajmone-Marsan  & F. Nonnis Marzano 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2005,122(1):62-68

The amplified fragment‐length polymorphism (AFLP) technology is a recently introduced method to investigate genomes of different complexity, from microbial to higher organisms. It is applied to purposes as diverse as identification of species, strain and varieties, investigation of genetic diversity within and between populations, simple and complex trait mapping, and construction of linkage and physical maps. This technology has been designed on the use of primers labelled with radioactivity and on AFLP fragment separation on sequencing gel. We show that the original EcoRI/TaqI AFLP protocol does not perform appropriately when transferred to fluorescent labelling and capillary electrophoresis (CE), and propose an improved protocol for the production of high‐quality AFLP markers in fish, rodents and artiodactyles by means of the Beckman‐Coulter CEQ2000 automatic DNA sequencer. In addition, we describe the procedure routinely used in our laboratory to obtain binary matrices from AFLP profiles with the aid of Genographer free‐share software (vers. 1.6.0, J.J. Benham, Montana State University), able to elaborate original fragment data and convert them to standard graphical formats for phylogenetic analyses. Comparison with radioactive AFLPs in goats confirmed the reliability of the protocol developed for CE. In fact, 107 fragments generated by two primer combinations and identified by both techniques were attributed the same scoring. Compared with traditional methods, the use of capillary systems and automated analysis increases data throughput and scoring reliability, decreasing the overall experimental error.  相似文献   

Protein content of equine seminal plasma and sperm plasma membrane in subfertile stallions     

P.N. Guasti  F.F. SouzaC. Scott  F.P. HartwigP.M. Papa  G.A.M. dos SantosC.P. Freitas-Dellaqua  F.O. Papa 《Journal of Equine Veterinary Science》2014

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HPLC quantification of coumarin in bastard balm (Melittis melissophyllum L., Lamiaceae)     

Maggi F  Barboni L  Caprioli G  Papa F  Ricciutelli M  Sagratini G  Vittori S 《Fitoterapia》2011,82(8):1215-1221

Coumarin occurs in many plants used as flavoring and is known to possess hepatotoxic effects. Despite in the EFSA ‘Compendium of botanicals containing toxic substances’ coumarin is reported to be present in Melittis melissophyllum (bastard balm), a plant traditionally used as beverage in Italy and Serbia, to the best of our knowledge quantitative data has never been reported. Thus, the amount of coumarin in bastard balm leaves and its variation during the annual phenological cycle were determined. The subsp. melissophyllum resulted to contain high levels of coumarin (14,392 mg/kg), mainly in the early stages of the plant cycle, suggesting prudence in its use as beverage. Furthermore, coumarin was found to be useful as marker compound to differentiate the bastard balm subspecies occurring in Italy, since the subsp. albida contained a much lower content of this molecule (19–34 mg/kg).  相似文献   

Influence of irrigation on the distribution and control of the nematode Meloidogyne javanica by the biocontrol bacterium Pasteuria penetrans in the field     

Kounbobr Roch Dabiré  Saliou Ndiaye  Jean-Luc Chotte  Sabine Fould  Mamadou Thiam Diop  Thierry Mateille 《Biology and Fertility of Soils》2005,41(3):205-211

Pasteuria penetrans, a bacterial parasite of plant-parasitic nematodes, is used to control root-knot nematode Meloidogyne spp. populations in vegetable crops. But its efficiency is variable, mostly because of the patchy distribution of the bacteria in arable fields. As the infective P. penetrans are non-motile bacteria in soil, abiotic soil factors can affect the bacteria–nematode relationships. An epidemiological study, conducted in a vegetable field, showed that abiotic factors such as irrigation, soil water holding capacity and texture, affected the efficiency of P. penetrans. A correspondence analysis between these abiotic factors and the density of P. penetrans spores in the soil, and the proportion of Meloidogyne javanica juveniles infected by the bacteria, revealed that irrigation affected directly the distribution of the spores in soil pores related to their passive transport by water flow. Laboratory experiments conducted on the passive transport of spores confirmed that intensive irrigation leached the spores down the soil profile and decreased the percentage of infected Meloidogyne juveniles.  相似文献   

Litter-fall and litter decomposition in a low Mediterranean shrubland   总被引：1，自引：0，他引：1  

Antonietta?FiorettoEmail author  Stefania?Papa  Amodio?Fuggi 《Biology and Fertility of Soils》2003,39(1):37-44

Annual production of litter by Cistus incanus (L.) and Myrtus communis (L.) and decomposition dynamics of leaf litter of these species was studied in a Mediterranean shrubland. Myrtus and Cistus produced 472 and 429 g dry weight litter m^-2 year^-1, respectively. Leaves were the predominant litter component for both species. The average decay constant of Myrtus and Cistus litters enclosed in litter bags, calculated over the whole study period (38 months), was 0.71 year^-1 and 0.31 year^-1 respectively. In green leaves the N content differed during growth seasons for both species, whereas the content of Ca, Mg, P, K, and Na did not show significant changes. Abscised leaves had lower N, P and K contents than green leaves, evidencing that a nutrient translocation before abscission occurred from senescent leaves. The nutrient contents of the leaves at abscission time, generally higher in Cistus than in Myrtus, allowed us to estimate the annual nutrient input to the soil. Phosphorus and K more than N were rapidly released by the decomposing litters after exposure. Nutrient limitation, in particular P, might be considered the main growth limiting factor for Myrtus and Cistus. Both species were adapted to recovery and rapidly recycle P more than N and K in the living biomass through retranslocation from green leaves before abscission and/or a high release rate from the decomposing litter. The former strategy was better used by Cistus, the latter by Myrtus.  相似文献   

Variation in the distribution of monosaccharides in soil fractions in the mounds of termites with different feeding habits (Senegal)     

Saïdou Sall  Alain Brauman  Saliou Fall  Corinne Rouland  Edouard Miambi  Jean-Luc Chotte 《Biology and Fertility of Soils》2002,36(3):232-239

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Testicular fine needle aspiration cytology from a stallion with testicular degeneration after external genitalia trauma     

Frederico O. Papa DVM  PhD  Denise P. Leme DVM  MS  PhD FAPESP Program 《Journal of Equine Veterinary Science》2002,22(3):121

A seven-year-old Quarter Horse had a serious external genitalia trauma with severe swelling of ventrum, penis, prepuce and scrotum after falling over a fence. Appropriated treatment was rapidly started after clinical examination. During recovery period, the spermatogenesis assess by semen evaluation was not possible due to stallion's inability to ejaculate. Therefore, for testicular evaluation fine needle aspiration cytology (FNAC) was performed. The first FNAC showed a deviation of germ cell line towards immature cells, mainly by primary spermatocytes (59.5%) with very few late spermatids and spermatozoa (2.5% each), and an increased Sertoli cells/germ cells ratio (478/100), which characterized testicular degeneration. One month after the first FNAC, the second exam presented a drastic decrease of Sertoli cells/germ cells ratio (7/100) and marked increase of mature cell number, specially by early and late spermatids (50% and 24.5%, respectively). In this case, the results of both FNAC could demonstrate a partial recovery of spermatogenesis activity. Two months later, the stallion had mated two mares successfully and they became pregnant. In conclusion, the adequate treatment allowed a complete recovery of the stallion's reproductive function, and since semen collection was impossible during treatment, testicular FNAC showed to be an efficient diagnostic method for evaluating acute damage in the spermatogenesis.  相似文献   

Effects of Pentoxifylline on Equine Epididymal Sperm     

Priscilla N. Guasti  Gabriel A. MonteiroRosiara R. Maziero  MSc  Ian MartinBruno R. Avanzi  MSc  José A. Dellaqua Jr.Frederico O. Papa  PhD 《Journal of Equine Veterinary Science》2013

This study evaluated whether pentoxifylline (PTX) present in the flushing extender influenced the function of equine epididymal spermatozoa after recovery and after thawing. For this experiment, 58 testicles from 29 Brazilian Jumping Horses were used. Cauda epididymides of each stallion were separated and flushed with a skim milk extender, with or without 7.18 mM PTX and then subjected to the freezing process. Samples flushed with the extender containing PTX showed a significant increase in total motility, progressive motility, straight line velocity, curvilinear velocity, and percentage of rapid sperm immediately after the recovery of epididymal sperm and after 15 minutes of incubation at 37°C (P < .05). However, the presence of PTX in the flushing extender did not affect the post-thaw motility parameters or plasma membrane integrity (P > .05). The results of this study showed that the PTX present in the flushing extender improved motility parameters of recently recovered epididymal sperm and had no deleterious effects on plasma membrane integrity and freezability of equine epididymal sperm.  相似文献   

FSH up-regulates angiogenic factors in luteal cells of buffaloes     

L.A. Fátima  M.C. Evangelista  R.S. Silva  A.P.M. Cardoso  P.S. Baruselli  P.C. Papa 《Domestic animal endocrinology》2013

Follicle-stimulating hormone has been widely used to induce superovulation in buffaloes and cows and usually triggers functional and morphologic alterations in the corpus luteum (CL). Several studies have shown that FSH is involved in regulating vascular development and that adequate angiogenesis is essential for normal luteal development. Angiogenesis is regulated by many growth factors, of which vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) have an established central role. Therefore, we have used a combination of in vitro and in vivo studies to assess the effects of FSH on the expression of VEGF and FGF2 and their receptors in buffalo luteal cells. The in vivo model consisted of 12 buffalo cows, divided into control (n = 6) and superovulated (n = 6) groups, and CL samples were collected on day 6 after ovulation. In this model, we analyzed the gene and protein expression of FGF2 and its receptors and the protein expression of VEGFA systems with the use of real-time PCR, Western blot analysis, and immunohistochemistry. In the in vitro model, granulosa cells were collected from small follicles (diameter, 4–6 mm) of buffaloes and cultured for 4 d in serum-free medium with or without FSH (10 ng/mL). To induce in vitro luteinization, LH (250 ng/mL) and fetal bovine serum (10%) were added to the medium, and granulosa cells were maintained in culture for 4 d more. The progesterone concentration in the medium was measured at days 4, 5, and 8 after the beginning of cell culture. Cells were collected at day 8 and subjected to real-time PCR, Western blot analysis, and immunofluorescence for assessment of the expression of FGF2, VEGF, and their receptors. To address the percentage of steroidogenic and growth factor-expressing cells in the culture, flow cytometry was performed. We observed that in superovulated buffalo CL, the FGF2 system mRNA expression was decreased even as protein expression was increased and that the VEGF protein was increased (P < 0.05). In vitro experiments with granulosa cells showed an increase in the mRNA expression of VEGF and FGF2 and its receptors 1 and 2 and protein expression of VEGF, kinase insert domain receptor, FGF receptor 2, and FGF receptor 3 in cells treated with FSH (P < 0.05), in contrast to the in vivo experiments. Moreover, the progesterone production by FSH-treated cells was elevated compared with untreated cells (P < 0.05). Our findings indicate that VEGF, FGF2, and their receptors were differentially regulated by FSH in vitro and in vivo in buffalo luteal cells, which points toward a role of CL environment in modulating cellular answers to gonadotropins.  相似文献