Measurement of C‐reactive protein and Prostaglandin F2α Metabolite Concentrations in Differentiation of Canine Pyometra and Cystic Endometrial Hyperplasia/Mucometra

Canine pyometra is a dioestrus period disease in which systemic inflammatory response syndrome (SIRS) is a common outcome due to the response of the body to the bacterial infection. The purpose of this study was i) to differentiate canine pyometra and cystic endometrial hyperplasia (CEH)/mucometra by measuring serum C‐reactive protein (CRP) and prostaglandin F_2α metabolite (PGFM) concentrations in blood and ii) to compare serum concentrations of CRP and PGFM in bitches with a pathological uterus (pyometra or CEH/mucometra) to concentrations in bitches with a healthy uterus. Mean CRP concentrations were found significantly higher (p < 0.001) in dogs with pyometra compared to those with CEH/mucometra or healthy uterus. However, no statistical difference could be detected between the groups for mean PGFM concentrations. Mean white blood cell count (WBC), alkaline phosphatase (ALP) and total protein concentrations were found significantly higher (p < 0.001) in dogs with pyometra. Escherichia coli was the most frequently isolated microorganism from dogs with pyometra (64.3%). Edwardsiella spp. was detected in a single case of pyometra for the first time. In conclusion, our results demonstrate that serum CRP concentrations were increased in dogs with pyometra and thus we conclude that serum CRP concentration but not PGFM might be useful as a marker to differentiate a case of CEH/mucometra from pyometra in female dogs. To the authors' knowledge, this is the first report in which Edwardsiella spp. has been isolated in the canine uterus.     

Is Mechanically Induced Cystic Endometrial Hyperplasia (CEH) a Suitable Model for Study of Spontaneously Occurring CEH in the Uterus of the Bitch?

Cystic endometrial hyperplasia (CEH) was experimentally reproduced by the intraluminal insertion of a silk wire in the uterus of metestral bitches to obtain a model for the study of spontaneously occurring CEH in the bitch. This mechanically reproduced CEH corresponded histologically very well to spontaneously occurring CEH. With respect to sex hormone receptor expressions, however, there was no similarity. In the mechanically induced CEH, progesterone receptor expression was reduced in the epithelial cells (surface epithelium and endometrial glands) and slightly increased in the stromal fibroblasts and myometrium as compared with the normal metestrus uterus. The oestrogen receptor expression in mechanically induced CEH was reduced in the epithelial cells (surface epithelium and endometrial glands) and more or less unchanged in the stromal fibroblast and myometrium as compared with the normal metestrus uterus. This is in contrast to the increased sex hormone receptor expressions in all uterine cell types, observed in spontaneously occurring CEH. The mechanically induced CEH model corresponds immunohistochemically more to early placentation and to pyometra in the bitch. Thus this experimental CEH model is not suitable for pathogenic studies of spontaneously occurring CEH in the bitch.     

Ultrasonography and Cystic Hyperplasia–Pyometra Complex in the Bitch

Cystic endometrial hyperplasia-pyometra complex is the most frequent and important endometrial disorder encountered in bitches. The pathogenesis of the disease is related to the activity of progesterone [Feldman and Nelson, Canine and Feline Endocrinology and Reproduction (1996) W.B. Saunders, Philadelphia]. Cystic endometrial hyperplasia (CEH) is an abnormal response of the bitch's uterus to ovarian hormones [De Bosschere et al. Theriogenology (2001) 55, 1509]. CEH is considered by many authors to be an exaggerated response of the uterus to chronic progestational stimulation during the luteal phase of the oestrous cycle, causing an abnormal accumulation of fluid within the endometrial glands and uterine lumen (De Bosschere et al. 2001). The resulting lesions of pyometra are due to the interaction between bacteria and hormones. The aim of this study was to evaluate if transabdominal uterine ultrasonography can be a useful and reliable diagnostic method to confirm Dow's [Veterinary Record (1958) 70, 1102] and De Bosschere's histopathological classification of CEH-pyometra complex. The study was carried out on 45 bitches with pyometra, 10 purebreds and 35 crossbreeds, 1-15 years old, 20% of which had whelped at least once. None of these animals had received exogenous oestrogen or progesterone treatment. On admission the 45 animals were in the luteal phase of the oestrus cycle. Clinical signs, blood parameters, uterine ultrasonography, bacterial swabs and uterine histopathological results were recorded. Results suggest that ultrasonographic examination is a useful and reliable tool for the diagnosis of cystic endometrial hyperplasia.     

ULTRASONOGRAPHY AND RADIOGRAPHY OF THE CANINE POSTPARTUM UTERUS

A vulvovaginal discharge following parturition in a bitch is often a cause of concern to owners and clinicians, especially if whelping was complicated in any way. Ultrasonography could potentially distinguish between normal and abnormal postpartum uterine states because the uterine wall and luminal contents can be imaged in detail. Five normal bitches were examined to determine the normal ultrasonographic appearance of the postpartum uterus and the sensitivity of ultrasonography in detecting the involuting uterus, comparing this sensitivity with that of radiography. Ultrasonography was done at 1, 4, 8, 12, 18 and 24 days postpartum, radiography at 1, 4, 8, 12 and 18 days postpartum. By 12 days postpartum, the uterus could not be seen on radiographs, while at 24 days postpartum the uterus was still sonographically identifiable. The ultrasonographic characteristics of the involuting uterus are described.     

产后奶牛子宫内膜的扫描电镜观察

应用扫描电镜观察了 5头奶牛产后不同时期子宫内膜的变化。结果发现 ,分娩后早期 ,奶牛子宫内膜部分破损 ,与破损部位相邻的细胞在产后 1周内相继脱落 ;未破损内膜上皮细胞表面的纤毛和微绒毛形状不整、数量减少。产后2周 ,基底细胞开始生长 ,至产后 4 5 d时生长完成 ;产后 4周 ,内膜破损区域缩小 ,上皮细胞表面纤毛和微绒毛形状规则、数量增加 ;产后 4 5 d,偶见上皮细胞缺失 ;至产后 6 0 d,子宫内膜上皮细胞完整、排列有序、被覆纤毛和微绒毛。上述结果表明 ,奶牛子宫内膜在分娩后 6 0 d内完成修复。     

Age‐Related Pregnancy Results and Further Examination of Bitches after Aglepristone Treatment of Pyometra

The cystic endometrial hyperplasia and pyometra complex is one of the most common uterine diseases in bitches. The appearance of pharmacological preparations containing anti‐progestagens created new possibilities for pyometra treatment. The aim of this study was to evaluate the curative effect of the anti‐progestagen aglepristone treatment of pyometra in bitches of different ages. Twenty four bitches of different breeds, aged from 0.8 to 9.5 years (21–48 kg) exhibiting clinical pyometra symptoms (two groups – I ≤ 5 years, n = 14 and II >5 years, n = 10) were evaluated. Information about the general reproductive health was collected up to 54 months after anti‐progestagen treatment. Remission of clinical symptoms and return of blood chemistry results and total leucocyte count to referential values were achieved in all cases within 14 days of treatment. Bitches were naturally mated at the first, and when unsuccessful, the second oestrus after treatment. In group I, no recurrence of pyometra symptoms was observed during following cycle(s). Eight bitches (57.1%) had a full‐term pregnancy and the number of newborn pups ranged from 1 to 12. None of the bitches from the group II became pregnant. In conclusion, the basic indication for conservative pharmacological treatment of pyometra is preserving female fertility and obtaining offspring. The important conditions for successful aglepristone treatment are: the young age (up to 5 years) and the lack of detectible ovarian cysts. It seems necessary to mate bitches in the first or second oestrus after finishing treatment. The efficacy of treatment can be measured by the after‐treatment pregnancy rate.     

Evaluation of cervical and uterine size,at 4 weeks postpartum,as a predictor of subsequent fertility in Jersey cattle

Uterine and cervical size of Holstein dairy cows is reported among reasons for a decline in dairy cow fertility. Therefore, the objectives of this study were to (a) determine whether size of the cervix and uterus at 4 weeks postpartum impacted subsequent fertility at first service in Jersey cattle, (b) determine whether progesterone level at 4 weeks postpartum impacted cyclicity and (c) the association of the presence of corpus luteum and uterus and cervix size. Body condition scores at calving, presence of postpartum diseases, parity number and milk weights were taken from lactating Jersey dairy cows (N = 147) for 28 days postpartum. During the fourth week postpartum, a blood sample was obtained for progesterone concentration, and transrectal ultrasonography was performed by a high-resolution ultrasound machine to determine cervical and uterine horn diameter, as well as ovarian structures measurements. Correcting for parity number, BCS at calving, presence of diseases and milk yield, cows with a cervix >2.54 ± 0.63 cm and uterine horn >2.25 ± 0.59 cm were less likely to become pregnant at first service (p = .04 and p = .003, respectively). The cows with larger cervix had a trend to be less likely to have a corpus luteum present at the 4th week of lactation (p = .067). Cows with larger uterine horn size were less likely to have a corpus luteum present at the 4th week of lactation (p = .015). It is concluded that a larger cervix and/or uterus during the postpartum was associated negatively with fertility and cyclicity in Jersey cows.     

The development of cystic endometrial hyperplasia in the bitch following serial uterine biopsies

Serial uterine biopsies were performed on two bitches during three oestrous cycles. The uterus was biopsied once in one other bitch and twice in another case following exteriorization and palpation of the uterine horn on four occasions earlier in the cycle. Following the serial biopsies both bitches developed a vulval discharge for a duration of 8-21 days. This first appeared between metoestrus days 14 and 30. By the 5th week after oestrus the uteri of both bitches developed cystic endometrial hyperplasia with inflammatory cell infiltration which regressed spontaneously. This lesion did not develop in the two other bitches, and it may have been caused by the surgical interference with the endometrium during the period of luteal development.     

Escherichia coli strains isolated from the uterus and urinary bladder of bitches suffering from pyometra: comparison by restriction enzyme digestion and pulsed-field gel electrophoresis.

Pyometra (uterine inflammation with accumulation of pus in the uterus) is regarded as one of the most common illnesses in bitches. The ethiology and pathogenesis are complex with both hormonal and bacterial elements. The bacteria most frequently isolated from the uterine content is Escherichia coli.In this study, 84 E. coli strains from the uteri of 70 bitches suffering from the disease were examined and their DNA-profiles compared by restriction enzyme analysis and pulsed-field gel electrophoresis (PFGE). Through variations in DNA-profiles of the E. coli isolates, this study indicates that pyometra is caused by E. coli originating from the normal flora of the dogs and not by certain clones spread between animals.E. coli strains from the urinary bladder and the uterus of six of the bitches suffering from simultaneous urinary tract infection and pyometra were examined and compared as above. The DNA-profiles of the isolates from each of the six bitches were 100% identical. This study supports the theory suggesting that in cases of simultaneous urinary tract infection and E. coli pyometra, the urinary tract and uterus are infected with the same bacterial strain.To evaluate whether the uterus was infected with a single clone of E. coli or if multiple clones were present, eight to 16 colonies of E. coli isolated from pyometra samples from a further 10 bitches were examined. All bacterial colonies from the culture of the same bitch showed identical DNA-profiles.In 14 of the 70 bitches, two macroscopically different but biochemically identical E. coli colony types were isolated. The two colony types from the same bitch proved to have identical DNA-profiles in 13 cases and almost identical in the remaining bitch.     

Prostaglandins and reproduction in female farm animals

Prostaglandins impact on ovarian, uterine, placental, and pituitary function to regulate reproduction in female livestock. They play important roles in ovulation, luteal function, maternal recognition of pregnancy, implantation, maintenance of gestation, microbial-induced abortion, parturition, postpartum uterine and ovarian infections, and resumption of postpartum ovarian cyclicity. Prostaglandins have both positive and negative effects on reproduction; they are used to synchronize oestrus, terminate pseudopregnancy in mares, induce parturition, and treat retained placenta, luteinized cysts, pyometra, and chronic endometritis. Improved therapeutic uses for prostaglandins will be developed when we understand better their involvement in implantation, maintenance of luteal function, and establishment and maintenance of pregnancy.     

Prostaglandin F2 alpha treatment of canine pyometra

Immediate and long-term outcomes of prostaglandin F2 alpha treatment for canine pyometra were studied in 10 bitches. Examination of pretreatment uterine biopsy specimens, taken for histopathologic diagnosis and classification of disease severity, revealed either type III or IV pyometra. Dinoprost tromethamine (0.25 or 0.5 mg/kg of body weight, SC) was given once daily for 3 days. Bitches were bred at the first posttreatment estrus and monitored for a minimum of one year. When pure cultures of Escherichia coli (n = 3) or Staphylococcus aureus (n = 1) were obtained from the vagina, these bacteria also were found in the uterus. Pretreatment WBC counts often did not reflect the severity of histopathologic findings in the uterus, but posttreatment WBC counts were useful in monitoring response to treatment. Four bitches produced a litter within one year of treatment. Four bitches (40%) had recurrence of pyometra within one year of treatment, and these same bitches had another recurrence after an additional prostaglandin treatment. Three additional bitches had a recurrence by 27 months after therapy, establishing a total recurrence rate of 77% (7/9). Results suggested that subclinical disease may persist after treatment, with clinical recurrence during diestrus. Despite the high recurrence rate, it was concluded that this treatment is a practical treatment for canine pyometra when reproduction is desired.     

Quantity of IL‐2, IL‐4, IL‐10, INF‐γ, TNF‐α and KC‐Like Cytokines in Serum of Bitches With Pyometra in Different Stages of Oestrous Cycle and Pregnancy

The occurrence of the pyometra is most common in the first half of the dioestrus when there is decreased cellular immunity associated with increased serum concentration of progesterone in females. The aim of this study was to determine the immunological profile of bitches with pyometra, studying serum levels of IL‐2, IL‐4, IL‐10, IFN‐γ, KC‐like and TNF‐α and comparing them with those of healthy bitches in anoestrus, dioestrus and pregnant. Forty females were divided into four experimental groups: group 1 (G1): with pyometra (n = 10); group 2 (G2): bitches in the second week of gestation (n = 10); group 3 (G3): in anoestrus (n = 10); and group 4 (G4): in dioestrus (n = 10). The serum levels for IL‐2, KC‐like, INF‐γ and TNF‐α were similar for all experimental groups. The values obtained for IL‐10 were found increased (p < 0.001) in animals in dioestrus and pyometra compared with females in anoestrus and pregnant, and the levels of IL‐4 observed were significantly greater (p < 0.001) in bitches with pyometra when compared with others. The cytokine profile in animals with pyometra is similar to bitches in dioestrus for IL‐10 and had increase in IL‐4 for bitches with pyometra, which represents an anti‐inflammatory these cases. This suggests the presence of an immunosuppressive state in both cases, which may explain the propensity of bitches in dioestrus to be affected by pyometra and the severity of the disease on these animals.     

Treatment of spontaneous pyometra in 22 bitches with a combination of cabergoline and cloprostenol

Twenty-two bitches with ultrasonographically diagnosed spontaneous pyometra were treated with a combination of 5 microg/kg cabergoline per day and 5 mug/kg cloprostenol every third day, and potentiated sulphonamide twice a day. Bitches with either open-cervix or closed-cervix pyometra showed a rapid clinical improvement, associated with a reduction in plasma progesterone concentration, increased vulval discharge and a reduction in the diameter of the uterus. The haematological profiles of 21 of the bitches returned to normal within six days of treatment, and their biochemical profiles returned to normal within nine days; 19 of the bitches were managed successfully by a 10-day period of treatment. Two bitches required a further three days of treatment, and in one bitch with a partial uterine torsion the treatment was not successful. Adverse effects of the treatment were limited to the 60 minutes immediately after the administration of prostaglandin, and included retching, vomiting, mild abdominal straining, diarrhoea and panting. The incidence of adverse effects was reduced after each successive dose of prostaglandin. Eleven of the 21 successfully treated bitches were mated at the next oestrus, and seven became pregnant; their litters were smaller than the published breed averages. In four of the bitches the pyometra recurred after the next oestrus.     

Efficacy and toxicity of estrogens commonly used to terminate canine pregnancy

Three groups of bitches were treated with diethylstilbestrol (75 micrograms/kg) orally for 7 days (n = 12), estradiol cypionate intramuscularly once (22 micrograms/kg; n = 12), or estradiol cypionate intramuscularly once (44 micrograms/kg; n = 12). Treatments commenced during late proestrus (n = 4/group), the fourth day of behavioral estrus (n = 4/group), or the second day of diestrus (n = 4/group). All bitches were bred on alternate days throughout estrus to stud dogs of known fertility. Ovariohysterectomies were performed on day 25 of diestrus to diagnose pregnancy and to assess any pathologic changes in the uterus. Eleven bitches treated with diethylstilbestrol, 6 bitches treated with the low dosage of estradiol cypionate, and 4 bitches receiving the high dosage of estradiol cypionate were pregnant at the time of surgery. Ten of the bitches treated with estrogens during proestrus, 6 treated during estrus, and 4 treated during diestrus were pregnant. The serum concentration of progesterone in 2 bitches treated with the high dosage of estradiol cypionate decreased to less than 2 ng/ml by day 25 of diestrus, suggesting premature luteal regression. Diethylstilbestrol appeared to have little efficacy in terminating pregnancy. Estradiol cypionate appeared to have greater efficacy when administered during estrus or early diestrus; however, pyometra developed in 2 bitches treated with this estrogen during diestrus.     

Uterine morphology and function in postpartum mares

Ultrasonically detectable characteristics of the uterus and embryo and palpable uterine tone were assessed in 10 postpartum mares. A bright fern-like pattern of ultrasonic uterine echogenicity, outlining the endometrial folds, was observed for an average of 2.1 ±0.2 days following parturition (range, 1 to 3 days). Unexpectedly, the uterus was quiescent throughout the postpartum interval, based on daily one-minute contractility scans. Contractility was maximal on Days 12 to 15 of pregnancy in both postpartum (n=7) and nonparturient (n=7) mares. The mean diameter of ultrasonically detectable intrauterine fluid collections increased (P<0.05) abruptly between days 1 and 2 postpartum and gradually decreased (P<0.05) between days 4 and 7; no collections were detected after day 16. There was no effect of day on echogenicity of the intrauterine fluid collections; on all days, fluid was relatively black or nonechogenic, suggesting that puerperal endometritis was not a problem in this group. Because the increase in intraluminal fluid occurred after parturition and in temporal association with a decrease in diameter and tone of the uterus, the fluid collections apparently represented a physiologic influx from the involuting uterus rather than residual placental fluid. Involution of the horns was completed by day 27 (formerly nongravid horn) and day 31 postpartum (formerly gravid horn), based on failure to detect further significant decreases in diameter. However, the formerly gravid horn was larger (P<0.05) in diameter than the formerly nongravid horn on each of Days 1 to 35 postpartum (end of experiment), indicating residual effects on uterine size. When averaged over both horns, uterine diameters were larger on Days 0 to 24 (Day 0=day of ovulation) of pregnancy in postpartum mares than in nonparturient mares; by Day 25, diameters were similar between statuses. By approximately Day 6 of pregnancy, uterine contractility and ultrasonic endometrial exhotexture were similar between postpartum mares and nonparturient mares. Uterine tone was greater (P<0.05) in postpartum mares than in nonparturient mares on all days between Day 0 and 25. An unexpected, transient increase in uterine tone was detected on Day 5 of pregnancy in both postpartum mares and nonparturient mares. No differences were found between reproductive statuses in patterns of embryo mobility, the day of fixation of the embryonic vesicle (postpartum, Day 15.3 ±0.4; nonparturient, Day 15.0 ±0.3), and diameter of the embryonic vesicle on the day of fixation (postpartum, 22.1 ±1.4 mm; nonparturient, 19.4 ±l.6mm). However, mean uterine tone and mean horn diameters on the side of fixation were greater (cranial and middle cornual segments; P<0.05) or tended to be greater (caudal segment; P<0.1 ) on the day of fixation in postpartum mares than in nonparturient mares. In all postpartum mares, fixation occurred in the formerly nongravid horn. Enhanced uterine tone in postpartum mares may account for the occurrence of fixation on the same day for the two reproductive statuses, despite the larger uterus in postpartum mares.     

Supplemental l-Arginine Shortens Gestation Length and Increases Mare Uterine Blood Flow before and after Parturition

Supplementing diets with l-Arginine (Arg) improves female reproductive performance and reproductive blood flow in other species. The objectives of this study were to investigate uterine artery blood flow changes before and after parturition, and evaluate blood flow in Arg supplemented and control mares by Doppler ultrasonography. Sixteen light-horse mares began Doppler ultrasonography evaluation, 21 days before expected foaling date (EFD) and continued until day 7 postparturition. The mares under treatment (n = 8) were supplemented with 100 g Arg, once daily, beginning with 21 days before EFD. Blood flow measurements were calculated as pulsatility index (PI) and resistance index (RI) for both uterine arteries, either ipsilateral or contralateral; to uterine horn of established pregnancy; defined gravid uterine artery (GUA) and non-gravid uterine artery (NGUA), respectively. The mares under treatment had a shorter gestation length (337 ± 1.7 days) as compared to control (345 ± 2.1 days; P ≤ .05). No differences in gestation length were observed between groups when examined by age, parity, EFD, or sex of foal. Both GUA and NGUA uterine artery diameter decreased from the day before parturition to day 7 after parturition (P ≤ .001). During this time period, both PI and RI increased (P ≤ .01); indicating less blood flow. A treatment effect was observed with Arginine-treated mares having greater blood flow prepartum in the NGUA (P ≤ .001) and postpartum in the GUA (P ≤ .05), for both indices. The data demonstrated that supplementing mares with Arg shortened gestation length and increased uterine arterial blood flow before and after parturition.     

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.     

Histological observations of canine cystic endometrial hyperplasia induced by intrauterine scratching

Of eight mongrel bitches, the antimesometrial side of the nonpregnant left horn of the uterus at the pregnant or nonpregnant luteal phase was scratched with a Kirschner's wire. Cystic endometrial hyperplasia (CEH) was induced in seven of the eight bitches (87.5%). No difference in the incidence of CEH in the left horn was seen between the pregnant and the nonpregnant groups. Histological examinations showed CEH with a dilatation of the basal glands, resembling "Swiss cheese endometrium".     

Ultrasound evaluation of pregnancy and spontaneous embryonic resorption in the bitch

One hundred and one pregnant bitches were imaged using real-time B-mode ultrasonography. The accuracy of predicting fetal number was 38 per cent during early pregnancy and was lower with later examinations. If bitches were examined in early pregnancy and estimated to have five pups or more, or four pups or less, the predictive value of ultrasound was 100 per cent and 83-3 per cent, respectively. The ultrasonographic appearance of one case of abortion and five cases of isolated spontaneous embryonic resorption were characterised. The resorptions did not apparently interfere with adjacent remaining conceptuses. The components of embryonic resorption were found to be; reduced volume and changes in echogenicity of embryonic fluid, loss of embryonic mass and heartbeat, collapse of the conceptus with thickening and inward bulging of the uterine wall and reduced size in comparison with adjacent conceptuses. Following resorption the uterus was moderately hypoechoic in appearance, similar to after parturition. No risk factors were correlated with resorption or abortion.     

Plasma concentrations of folic acid, vitamin B12 and progesterone of cyclic bitches, bitches during pregnancy and induced abortion and bitches with pyometra

The aim of this study was to investigate the plasma concentrations of folic acid, vitamin B12 and progesterone at different stages of the sexual cycle and pregnancy, during induced abortion and in bitches with pyometra. Bitches (n = 97) were assigned to groups as follows: a) oestrous cycle (n = 42) b) pregnancy (n = 25) c) induction of abortion (n = 10 and d) pyometra (n = 20). Oestrous cycle stages were determined by vaginal inspection and cytology. Pregnancies were estimated by ultrasound (5.0 Mhz; linear transducer; Schimadzu) at days 15-25, 35-45 and 46-63 of pregnancy. Treatments for the induction of abortion were started between days 25 and 35 after mating (5 microg/kg cabergoline daily, Galastop; 5-10 microg/kg Alfaprostol every other day, Gabbrostim). Diagnosis of pyometra was confirmed by ultrasound and vaginoscopy. Folic acid and vitamin B12 concentrations did not differ among different stages of the oestrous cycle. The mean concentration of folic acid during early pregnancy (days 15-25) exceeded levels of later stages (days 46-63): 9.4 +/- 3.7 microg/ml and 4.7 +/- 1.8 microg/ml, respectively (p < 0.01). A positive correlation between folic acid and vitamin B12 was determined in pregnant dogs ( r = 0.925; p < 0.02). Before the induction of abortion, the concentration of folic acid was 9.6 +/- 5.2 microg/ml; during abortion it decreased to 5.0 +/- 3.2 microg/ml (p < 0.01). A significant correlation (r = 0.925; p < 0.02) between progesterone and folic acid was obtained in bitches with abortion. The mean concentration of folic acid in bitches with pyometra significantly differed from that of bitches at different stages of the oestrous cycle (p < 0.05). The mean concentration of folic acid was significantly lower in metoestrous bitches when compared to bitches with pyometra (p < 0.05). The decrease of serum concentrations of folic acid during pregnancy and induced abortion show that fetal growth and abortion caused higher consumption of folic acid. Concerning bitches did not show any deficiency symptoms, which is why it can be concluded that this decrease is physiological.