An immunohistochemical study of ovarian follicle histogenesis in the early post-hatch Japanese quail (Coturnix coturnix japonica)

The early post-hatch development of immunoreactivity to vimentin, desmin, smooth muscle actin (SMA) and laminin, in relation to follicle histogenesis, was described in this study. Ovigerous cords in day old quails contained pre-granulosa cells and oocytes. Pre-granulosa cells at the cortico-medullary junction were vimentin immunopositive. A laminin immunopositive basement membrane and desmin immunopositive mesenchymal cells lined the ovigerous cords. Ovigerous cords in 3-day-old quails contained developing primordial follicles, the vimentin immunopositive pre-granulosa cells of which were partially encircled by a basement membrane and desmin immunopositive mesenchymal cells. In 5- to 7-day-old quails, ovigerous cords formed an outer cortical region, while primordial follicles formed the inner cortical region. Early pre-vitellogenic follicles were present in 9- to 13-day-old quails. Underlying the granulosa cells of these follicles was a laminin immunopositive basement membrane and a layer of desmin immunopositive thecal cells. Early and late pre-vitellogenic follicles dominated the ovary in 15- to 17-day-old quails. The thecal layer in these follicles was desmin immunopositive, but SMA immunonegative. The results of the study have shown that the process of primordial follicle development in the Japanese quail is similar to that reported in mammals. The study suggests that in the quail pre-granulosa cells originate predominantly from the medulla. The study has shown that, in the Japanese quail, thecal cells are derived from desmin immunopositive mesenchymal cells lining the ovigerous cords.     

Ultrastructural features of the follicular wall in developing follicles of the sexually immature ostrich (Struthio camelus)

The ultrastructure of the follicular wall in primordial, previtellogenic and vitellogenic follicles of the sexually immature ostrich is described in the present study. The follicular wall consists of a zona radiata, granulosa cell layer, basal lamina and thecal layer. Cytoplasmic processes from the plasma membranes of the granulosa cell layer and the ovocyte form the zona radiata in previtellogenic and vitellogenic follicles. The granulosa cell layer transforms from simple cuboidal epithelium in primordial follicles to simple columnar or pseudostratified columnar epithelium in previtellogenic and vitellogenic follicles. Transosomes were observed along the apical and lateral plasma membranes of granulosa cells. The thecal layer in previtellogenic and vitellogenic follicles consists of interna and externa components. The fibroblasts in the theca externa contain microfilaments, which are thought to be actin filaments. The study revealed ultrastructural features, which are associated with the transportation of yolk precursors and nutrients into the ovoplasm. In addition, the study indicates that, although the cells in the theca externa contain microfilaments, they do not possess the ultrastructural characteristics of smooth muscle cells.     

Structural and immunohistochemical features of the epididymal duct unit of the ostrich (Struthio camelus)

The epididymal duct unit, comprising the ductus conjugens, ductus epididymidis and ductus deferens, was studied histologically, ultrastructurally and immunohistochemically in five sexually mature and active birds. The main morphological features of the pre-dominant non-ciliated (type III) cell of the epithelial lining of this duct unit include, but are not limited to, a moderately abundant smooth or sparsely granulated endoplasmic reticulum, electron-dense secretory granules and numerous mitochondria in the supranuclear zone of the cytoplasm. A single, large heterogeneous lipid droplet, of unknown function, was characteristically situated immediately proximal to the nucleus. The epithelium is obviously secretory and specifically, of the merocrine, and not apocrine, type of secretion. The epithelium of the epididymal duct unit was only focally and weakly to moderately immunopositive to both actin MF and desmin IF, while the duct unit was immunonegative to cytokeratin and vimentin intermediate filaments. The peritubular muscular layer was moderately to strongly positive to both actin and desmin, and negative to cytokeratins and vimentin.     

Current attitudes to,and use of,peri-operative analgesia in dogs and cats by veterinarians in New Zealand

AIM: To investigate the attitudes of veterinary practitioners in New Zealand to pain and analgesia, and their use of analgesic drugs, in dogs and cats.

METHODS: A questionnaire posted to 1,200 practising veterinarians was used to gather information about the use of analgesia in dogs and cats, assessment of pain, attitudes to pain relief, analgesic drugs and procedures used, factors affecting choice of analgesic agent, and veterinary demographics, continuing education and staffing.

RESULTS: Three hundred and twenty questionnaires with useable data were returned, a response rate of 28%. Male and female veterinarians were evenly represented. The analgesic agents most commonly used were morphine (opioids) and carprofen (a non-steroidal anti-inflammatory drug; NSAID). Use of peri-operative pain relief ranged from 50% for castration of cats to 91% for fracture repair in dogs. For most procedures, female veterinarians scored pain at a significantly higher level than their male colleagues. Fifty-eight percent of respondents considered their knowledge in the area of assessment and treatment of pain was adequate.

CONCLUSIONS: This survey was considered representative of veterinarians working in companion animal practice in New Zealand. Results indicated a relatively high use of peri-operative analgesia, including both pre-emptive and multi-modal analge- sia, in cats and dogs, although there was still some disparity between the perception of how painful a procedure was and the consequent use of pain relief.

CLINICAL RELEVANCE: The establishment of current attitudes and practices indicates to practising veterinarians how their own use of analgesics compares with that of their colleagues. It also provides information to educators on potential areas of focus, given that 42% of respondents felt their knowledge in the area of assessment and treatment of pain was inadequate.     

Activation of Mouse Oocytes Following Intracytoplasmic Injection of Chicken Sperm Extract

The cytosolic factor from the sperm of a wide variety of species has been reported to induce [Ca²⁺]i increase and/or activation in oocytes. Although many species have been studied, there is no data available on the chicken sperm factor responsible for activation of oocytes. This study was aimed to verify the activation of mouse oocyte by intracytoplasmic injection of chicken sperm extract (CSE). Survival rate of oocytes without rupture or lytic degeneration following injection was greater than 80% regardless of the presence or absence of CSE. Among the intact oocytes, activation rates following injection were 27.5% (11 of 40) in the sham operation group. Injection of CSE resulted in significantly higher activation rates in the 1/10 dilution group (57.8%, 26 of 45) compared with the sham operation group (p = 0.0045). Calculated amount of injected sperm extract of 1/10 dilution of CSE was equivalent of 12 chicken sperm. When the 1/10 diluted mouse sperm extract (MSE) was injected, survival rate of injected oocytes and activation rate of the survived oocytes was 82% (41 of 50) and 78% (32 of 41), respectively. Activation rate of MSE‐injected oocytes was a little but significantly higher than that of CSE‐injected oocytes (p < 0.037). In conclusion, it suggests that CSE can activate the mouse oocyte and that oocyte activating sperm factor(s) may be common between avian and mammal.     

Effects of Glucose Concentration on in vitro Fertilization in BALB/c Mice

BALB/c mice are widely used in genetic, tumour and immunological studies. However, the mice demonstrate a lower reproduction rate, low fertility and small litters, because of their highly genetic homozygoisty. Based on in vitro fertilization (IVF), a routine technique for biomedical studies, it is worth to evaluate the effects to BALB/c mice on IVF efficiency. In order to test the genetic factor affecting the IVF efficiency of BALB/c, four reciprocal IVF tests of BALB/cByJ and FVB/NCrl mice were performed. The results showed that the average fertility of IVF sponsored by FVB/NCrl spermatozoa was 69.6%, but only 12.1% was obtained from BALB/cByJ strain. Effect of glucose contained in the culture medium to the IVF efficiency of BALB/cByJ was also evaluated. The results showed that the fertility of BALB/cByJ spermatozoa incubated with 0, 2.7, 5.5, 11.1 and 22.2 mm of glucose in the TYH medium were 6.8, 9.9, 13.9, 32.7 and 22.2%, respectively. It is showed that IVF efficiency of BALB/cByJ spermatozoa could be improved depending on the concentration of glucose in the IVF medium. According to the results, it is beleived that lower IVF of BALB/cByJ mice might be due to the genetic defect in spermatozoa and increasing glucose in the IVF medium which significantly affect the IVF efficiency of BALB/cByl via activating the spermatozoa.     

Prolactin,Androstenedione and IGF1 Serum Concentrations During Induced Follicular Growth by eCG Administration in the Bitch

The oestrus cycle in the domestic bitch, a monoestrous species, differs considerably from that of other veterinary domestic animals species. In the bitch the combined use of eCG and hCG is effective to induce oestrus predictably and safely (Stornelli et al., Theriogenology, 78, 2012 and 1056). Although several studies were done to describe the hormonal changes during the canine oestrus cycle, to our knowledge none was done to describe the hormonal changes during induced follicular growth after the administration of eCG. The aim of this work was to study prolactin (PRL), insulin‐like growth factor (IGF1) and androstenedione (ANDR) serum concentrations during follicular growth induced by a single dose of eCG administered to late anoestrous bitches. PRL and ANDR concentrations were lower before than after eCG TRT (before eCG vs pro‐oestrus, oestrus and dioestrus; 4.3 ± 1.8 ng/ml vs 6.5 ± 1.6 ng/ml, p < 0.05; 0.08 ± 0.2 ng/ml vs 0.42 ± 0.16 ng/ml, p < 0.05). Conversely, IGF1 concentrations were similar before and after eCG TRT (286.0 ng/ml ±32.2, p > 0.53). Additionally, PRL concentrations were similar before oestrus compared to during oestrus and dioestrus (6.9 ± 1.7 ng/ml, p > 0.19). Furthermore, IGF1 concentrations were higher before and during oestrus compared to first day of dioestrus (286.1 ± 29.8vs 200.4 ± 29.2 ng/ml, p < 0.01). On the contrary, ANDR concentrations were lower before and during oestrus compared to first day of diestrum (0.35 ± 0.17 ng/ml and 0.38 ± 0.15 vs 0.68 ± 0.17 ng/ml, p < 0.05). These results show that treatment with a single injection of 50 IU/kg of eCG in late anoestrous bitches successfully induced changes in follicular growth which were paralleled with changes in PRL, IGF1 and ANDR serum concentration similar to those occurring during a normally occurring oestrous cycle. In addition, our results suggest that IGF1 in the bitch could play an important role in ovarian folliculogenesis.     

Doppler and Contrast‐Enhanced Ultrasonography of Testicles in Adult Domestic Felines

The objective was to characterize the vascular patterns of testicular blood flow of adult cats, measuring the systolic velocity (SV), diastolic velocity (DV), resistance index (RI), gate time (wash‐in) peak enhancement and output time (wash‐out) of the contrast and addition of tissue fill characteristics. Forty‐five adult cats were selected, and the echotexture, echogenicity, size, contours and margins of testicles were assessed via ultrasound. By Doppler were evaluated the blood flow and determined of vascular index in testicular artery (SV, DV and RI) and via contrast‐enhanced ultrasonography determine the time for phases: wash‐in, wash‐out and peak enhancement. Sonographic findings presented normal. Testicular artery was observed in the spermatic cord with tortuous patter and showed monophasic‐patterned waves and low vascular resistance and with systolic peak evident. Values of indices vascular were as follows: SV = 6.73 cm/s, DV = 2.8 cm/s and RI = 0.54 for left testicles; and SV = 6.23 cm/s, DV = 2.77 cm/s and RI = 0.53 for right testicles. Contrast filled the subcapsular vascular structures and after a few seconds, a homogeneous moderate enhancement of the parenchyma, with parenchymal vessels still distinguishable and after the peak phase, a rapid homogeneous decrease in echogenicity. Values of time for contrast‐enhanced ultrasonography were as follows: wash‐in = 8.78 s, peak enhancement = 21.62 s and wash‐out = 75.36 for left testicles; and wash‐in = 10.76 s, peak enhancement = 21.50 s and wash‐out = 81.81 for right testicles. Doppler and contrast‐enhanced ultrasonography of the testicles in healthy adult cats was easily implemented and may provide baseline data for this organ to allow the use of these techniques as a diagnostic tool for evaluating testicular abnormalities in sick cats.