The Effect of Detopping the Field Bean Plants (Vicia faba L. minor) and Lack of the Pod Setting on the Symbiosis with Root Nodule Bacteria

The plants of field bean var. Nadwiślaνski were detopped after about 1 month since the beginning of flowering and after that all flower buds were removed from some plants. The effect of a change in the trophic balance on the setting and subsequent growth of root nodules was studied.
The removal of pods had the greater effect on the increase in the weight of the remaining organs than the detopping of plants. Detopping retarded the ageing of the plants and delayed the phase of the greatest reduction of the number of pods. The detopping of plants revealed their considerable compensative possibilities in terms of production of the pod weight and the potential of leaves for biomass production. The data obtained indicate that plant detopping may disturb the root–bacteria exchange of signals, necessary to initiate the nodule development, while the further growth of nodules depends, above all, on the amount of available photosynthates.
A very high correlation between the dry weight of the whole plant to the dry weight of leaves ratio and the weight of nodules was observed. It proves the importance of the trophic balance of the plant for the root nodule weight production.     

The Content of β-endorphin-like Immunoreactivity in Porcine Corpus Luteum and the Potential Roles of Progesterone, Oxytocin and Prolactin in the Regulation of β-endorphin Release from Luteal Cells in vitro

The amount of β‐endorphin‐like immunoreactivity (β‐END‐LI) in porcine corpora lutea from several stages of the oestrous cycle and the effects of progesterone, oxytocin, and prolactin on β‐END‐LI secretion in vitro by luteal cells were studied. Porcine corpora lutea obtained on days 1–5, 6–10, 11–13, 14–18, and 19–21 of the cycle were used to prepare extracts for β‐END‐LI determination. Additionally, corpora lutea from days 11–13 and 14–18 were enzymatically dissociated and isolated luteal cells were used for further study of β‐endorphin secretion in vitro. Cells were cultured in serum‐free defined M 199 medium (10⁶ cells/ml) at 37°C under 5% CO₂ in air, for 12 h. The influences of the following factors on β‐END‐LI secretion by luteal cells were tested: progesterone (10^–9, 10^–7 and 10^–5M ), oxytocin (0.01, 0.1, 1 and 10 ng/ml), and prolactin (0.1, 1, 10 and 100 ng/ml). The β‐END‐LI contents in extracts and media were measured by radioimmunoassay. The tissue concentration of β‐END‐LI was lowest on days 1–5 of the cycle (0.35 ± 0.03 ng/g wet tissue). Subsequently, it constantly increased to the highest value on days 14–18 (16.58 ± 0.52 ng/g wet tissue) and on days 19–21 it declined (11.10 ± 0.52 ng/g wet tissue). Progesterone at a low dose (10^–9 M ) resulted in significant (p < 0.05) increases and decreases in β‐END‐LI secretion by luteal cells from days 11–13 and 14–18, respectively. Higher doses of progesterone (10^–7 and 10^–5 M ) had no effect on β‐END‐LI release, compared with the control group. All dose‐levels of oxytocin used decreased β‐END‐LI secretion by luteal cells on days 11–13 and 14–18 of the cycle. Prolactin at doses of 0.1 and 1 ng/ml on days 11–13, and all doses tested on days 14–18 resulted in decreases in β‐END‐LI release from luteal cells. These results document evident changes in β‐END‐LI content in the pig corpus luteum during its development and indicate the potential roles of progesterone, oxytocin, and prolactin in luteal cell secretion of β‐END‐LI.     

Differences in Reproductive Performance, Embryo Development, Interferon-tau Secretion by the Conceptus and Luteal Function in Ewe Lambs Synchronized in Oestrus before or after the Spontaneous Onset of Luteal Activity Preceding Puberty

In mid-September, 1 month before the insertion of intravaginal pessaries to induce sexual activity, blood samples were collected every 4 days from 16 ewe lambs aged 7 months, in order to determine the incidence of ovulations by measurement of plasma progesterone concentrations. It has been studied whether the response to a progestagen treatment of ewe lambs apparently close to puberty could be modified by the onset of the ovarian events preceding puberty. The effect of the presence or absence of ovulations prior to progestagen treatment on the potential reproductive performance (fertility, litter size and fecundity), embryo development [embryo quality and interferon-tau (IFNτ) secretion], luteal function (progesterone secretion in vitro ) and endometrial progesterone content was studied in seven ovulating (Ov+) and nine nonovulating ewe lambs (Ov−) on day 14 after mating. The best potential reproductive results were obtained with Ov+ animals, although these differences could not be initially attributed to either different progesterone secretion in vitro or concentration of endometrial progesterone. Irrespective of the experimental groups, secretion of progesterone by luteal tissue from ewe lambs with normal embryos was significantly greater (p<0.05) than that of animals with abnormal embryos or with no embryos. Normal embryos secreted a higher amount of IFN-τ than those embryos classified as abnormal (p<0.07). In conclusion, ewe lambs which exhibit luteal activity before puberty have the highest levels of reproductive performances after a progestagen treatment. Corpora lutea from ewe lambs with normal embryos had higher rates of progesterone secretion in vitro and their embryos had a higher IFN-τ production by the embryos, indicating greater capacity for subsequent development.     

α‐6 Integrin Expression in Bovine Spermatogonial Cells Purified by Discontinuous Percoll Density Gradient

The study of spermatogonial stem cells (SSCs) provides a model to better understand adult stem cell biology. Besides the biomedical potential to perform studies of infertility in many species, SSCs hold a promising application at animal transgenesis. Because stem cells are thought to be associated with basement membranes, expression of α‐6 integrin has been investigated as a marker of type A spermatogonial cells, which are considered SSCs because of their undifferentiated status and self‐renewal ability. In this manner, the aim of this study was to isolate type A SSCs from adult bulls by a two‐step enzymatic procedure followed by a discontinuous Percoll density gradient purification and verify the expression of α‐6 integrin by flow cytometry and real‐time RT‐PCR before and after Percoll purification. Spermatogonial cells were successfully obtained using the two‐step enzymatic digestion. An average of 1 × 10⁵ viable cells per gram of testis was isolated. However, the discontinuous Percoll did not purify isolated cells regarding α‐6 integrin expression. Flow cytometry analysis demonstrated no differences in the α‐6 integrin expression between cell samples before and after Percoll purification (p = 0.5636). The same was observed in the real‐time PCR analysis (p > 0.05). In addition to α‐6 integrin, the expression of GFRa‐1 and PGP9.5, known bovine SSCs markers, was detected in all samples studied. Considering that Percoll can reduce cell viability, it is possible to conclude that Percoll density gradient is not suitable to purify bovine SSC, according to α‐6 integrin expression.     

Endometrial Adenocarcinoma and Mucometra in a 6-year-old Alaska Malamute Dog

A 6-year-old female Alaska Malamute dog was presented for evaluation of abdominal enlargement referred by a local veterinarian. On the history, the owner complained of chronic abdominal enlargement initiated more than 4 months ago, reduced appetite, occasional vomiting and general dullness. He also complained of greenish mucous intermittent vaginal discharge starting 10 days ago. The bitch was chronically treated with medroxiprogesterone acetate. A laparatomy was performed and fluid in the abdomen was found and aspirated during the surgery. Also a very fluid-filled distended uterus and a mass in the distal part of the left uterine horn were found. The mass was encapsulated by the omentum, but areas of necrosis and calcification were identified. Histopathological diagnosis was endometrial adenocarcinoma.     

Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.