Different fixative solutions in the detection of mast cells in the canine and feline reproductive organs.

BACKGROUND: The aim of the study was to evaluate the usability of different fixative fluids in the detection of mast cells in ovaries and uteri of female dogs and cats. MATERIALS AND METHODS: Samples were fixed in 4% formaldehyde, Carnoy's fluid, Mota's basic lead acetate and isotonic formaldehyde-acetic acid (IFAA). RESULTS: Mast cells (MCs) were detected by acidified toluidine blue staining and counted for various parts of the ovaries and uteri. In the ovaries of both species, the numbers of MCs were significantly (p < 0.05) higher in Carnoy than in formalin. No significant differences were found between Carnoy and Mota (tested only in cats). In the uterus, numbers of MCs were significantly (p < 0.05) higher in Carnoy, Mota and IFAA compared to formalin (canine endometrium, feline endometrium and feline myometrium), in Carnoy and Mota compared to formalin (canine myometrium) and in Mota compared to IFAA (feline myometrium). The majority of MCs were formalin-sensitive in the canine and feline uterus, in the canine ovary and in the feline cortex ovarii. In the feline medulla ovarii, the majority of MCs were formalin-resistant. No formalin-resistant MCs were detected in the feline tunica albuginea ovarii. CONCLUSIONS: Thus, using Mota's or Carnoy's fluid in the canine or feline female reproductive organs is recommended. This study improves methodology for all studies which clarify the role of MCs in the reproductive organs of the domestic and laboratory animals.

Distribution of mast cells in the feline ovary in various phases of the oestrous cycle.

This study is the first description of the distribution of mast cells in various phases of the oestrous cycle in the ovary of cat. Furthermore, this is the first description in species with an induced ovulation. The aim was to describe the distribution of mast cells and variability of their numbers in the feline ovaries in different phases of the oestrous cycle. The number of mast cells in medulla ovarii was affected by the estradiol and progesterone level in the blood serum because the lowest number was detected in anoestrus when the levels of hormones were basal. Nevertheless, both high and low numbers of mast cells were found in oestrus and dioestrus. To conclude, mast cells seem to be essential for the induction of spontaneous ovulation, but they do not play the same role for ovulation itself in cats with induced ovulation.

Azoospermia with variable testicular histology after 7 months of treatment with a deslorelin implant in toms.

The main aim of the study was to assess whether the longer use of a GnRH-agonist implant (deslorelin 4.7 mg, Suprelorin) in toms would lead to the suppression of spermatogenesis comparable with histologic appearance in juvenile animals as was previously described in dogs. The other aims were to monitor the progression of the testes size decrease and development of azoospermia 5 to 7 months after treatment with a GnRH-agonist implant. In animals, 5, 6, and 7 months after GnRH-agonist implant insertion, variable histological appearance of germinal epithelium was found, when tubules with elongating spermatids, round spermatids, spermatocytes, and spermatogonia as the most developed germinal cells were found in each group of toms. In all male cats, 5, 6, and 7 months after implant insertion, testosterone concentrations and testes size significantly differed between the first and the last visit. All animals, except one tom castrated 5 months after implant insertion, developed complete azoospermia. However, in this tom, all spermatozoa were immotile. Treatment with the subcutaneous GnRH-agonist implant was well tolerated, and no treatment-related adverse effects were noted. These results reported the efficacy of 4.7-mg deslorelin implant (Suprelorin) during its 7 months of use. The complete azoospermia confirms its contraceptive effect. However, the histologic evaluation revealed a great individual variability in the degree of spermatogenic suppression. The question as to whether spermatogenesis in toms can be suppressed in all males to the level of spermatogonia/primary spermatocytes after prolonged exposure to deslorelin has yet to be answered.

γδ T lymphocytes are recruited into the inflamed uterus of bitches suffering from pyometra.

Very little is known about the occurrence of immune system cells in the canine uterus. The aim of this study was to generate information about lymphocyte subsets that are present in the healthy canine uterus and that are recruited under inflammatory conditions caused by pyometra. Using immunohistochemistry and flow cytometry, a significant influx of γδ T lymphocytes was found in pyometra samples mainly due to recruitment of γδ(+)/CD8(-) T lymphocytes. The relative expression of genes encoding selected cytokines/chemokines was evaluated in samples from healthy and pyometra-affected uteri. Expression of pro-inflammatory cytokines (including IL-1ß, TNF-α, IL-8, IL-17 and IFN-γ) and chemokines (including CXCL10, CCL4 and CCL5) was upregulated in pyometra samples confirming the presence of inflammation. In contrast, the expression of the homeostatic chemokine CCL25 and of the anti-inflammatory cytokine IL-10 was downregulated and unchanged, respectively.

Reversible suppression of sexual activity in tomcats with deslorelin implant.

The aim of the study was to assess the efficacy of using a Gn-RH agonist implant (deslorelin, 4.7 mg, Suprelorin) to control sexual activity of male cats and reestablishment of sexual function after the implant removal 4 mo after placement. Using a control group (Group 1, n = 5), 22 domestic tomcats were given the implant subcutaneously in the region of the right shoulder blade and were then divided into two treatment groups. Animals in Group 2 (n = 14) were observed from the date of implant surgery and the observation lasted for 4 mo. In Group 3 (n = 8) all animals were monitored from the date of implant surgery. Then, after 4 mo, all implants were removed and the toms were observed for a further 4 mo. In all animals during their first visit and then in 1-mo intervals, changes in testosterone concentrations were assessed before (T0) and 4 h after (T4) human chorionic gonadotropin (HCG) administration and testis size was measured. In all tomcats, semen collection was performed, using an electroejaculator, in the course of the first visit and then in 2-mo intervals or at the end of observation. Total sperm count was determined in each semen sample. Two to four animals were castrated at weeks 4, 8, 12, 16, 20, 24, 28 and 32 and histologic assessment of the testes was performed. By evaluation of 200 cross sections of seminiferous tubules, the degree of spermatogenic suppression was assessed and animals in Groups 2 and 3 were assigned into groups according to most tubules with the most developed germ cell observed: G1, spermatocytes; G2, round spermatids; G3, elongating spermatids and G4, elongated spermatids. The mean area of Leydig-cell nuclei was calculated. In animals in Group 2, suppression after implant insertion was monitored. T4 concentrations, testis size, and total sperm count gradually decreased (P < 0.01; P < 0.01; and P < 0.05, respectively) within 4 mo after implantation. Histologic evaluation showed a high individual variation in the degree of suppression of spermatogenesis. In animals in Group 3, the implant was removed 4 mo after insertion and the return of sexual activity was monitored. Within 4 mo, T4 concentration and total sperm count increased to the physiological values of intact toms. Testes gradually increased in size and within 4 mo of implant removal almost reached pretreatment size. According to histologic evaluation of the seminiferous tubules, as early as 1 mo after implant removal, all animals were assigned to G4, with most tubules containing elongated spermatids as the most developed germ cells. Treatment with the long-term subcutaneous Gn-RH agonist implant was well tolerated and no adverse treatment-related effects were noted. These results demonstrated efficacy of 4.7 mg deslorelin implant (Suprelorin) with high variability of the effect onset in tomcats. Furthermore, the study revealed a strong need for complex examination, including testis size measurement, monitoring of hormonal changes, spermatological analysis and histologic evaluation, to declare the animal infertile. After the implant removal, all observed parameters confirmed the reversibility of the method and gradual return of sexual activity in toms.

Lactoferrin in canine sera: a pyometra study.

The concentration of lactoferrin was measured in canine sera from groups of healthy male dogs as well as pregnant and non-pregnant female dogs and was compared with that of bitches with pyometra. Lactoferrin concentrations were higher in bitches with pyometra. The role of elevated lactoferrin concentrations in the suppression of lymphocyte activity was examined in sera from bitches with pyometra in a series of investigations. Although the sera from bitches with pyometra were capable of suppressing lymphocyte activity, lactoferrin was not found to be involved in this action.

Hysterectomy leads to fast improvement of haematological and immunological parameters in bitches with pyometra.

OBJECTIVES: To investigate whether a combination of hysterectomy and antibiotic treatment leads to an improvement of altered haematological and immunological parameters in bitches affected by pyometra. METHODS: Blood samples obtained from 13 bitches affected by pyometra were investigated before hysterectomy and seven days later for a total and differential leucocyte count, activity of neutrophils and lymphocytes, and quantification of total serum immunoglobulins, lysozyme and circulating immune complexes. RESULTS: The parameters most affected included changes in blood profile (leucocytosis because of neutrophilia or leucopenia) and inhibition of lymphocyte activity. Seven days after hysterectomy, all affected parameters returned to normal levels comparable to clinically healthy dogs. CLINICAL SIGNIFICANCE: Application of immunostimulating therapy is not necessary to manage changes in blood cell counts and suppression of lymphocyte activity in bitches with pyometra. Removal of the infected uterus, being the source of infection, leads to improvement.