Blood calcium,glucose and haematology profiles of parturient bitches diagnosed with uterine inertia or obstructive dystocia

Bitches with dystocia most often present with clinical signs of uterine inertia (UI). The aetiology of myometrial dysfunction in most of these cases is still not elucidated. We compared blood ionized calcium (iCa) and glucose concentrations in bitches diagnosed with primary UI (PUI, n = 14), secondary UI (SUI, n = 6) or obstructive dystocia (OD, n = 6), and we described their haematology profiles. Bitches diagnosed with UI had a patent birth canal and delivered no puppies yet (PUI) or only part of the whole litter (SUI). The OD group had no UI and showed strong abdominal contractions. Blood iCa did not differ between the PUI, SUI and OD groups and was not influenced by litter size. There was a significant positive relationship (R² = .241, p = .013) between iCa concentrations and the dam's body weight. Glucose concentrations were also not significantly different between dystocia groups or influenced by body weight and litter size. Hypocalcaemia was detected in 11 bitches, and hypoglycaemia in two bitches. Pregnancy‐associated anaemia was seen in about one‐third of the bitches. Eight of 12 dogs had increased platelet counts, and ten had leukocytosis with mature neutrophilia. Although iCa did not differ between dystocia groups, low concentrations may have contributed to the development of UI in some of the small size bitches. Hypoglycaemia was uncommon, and therefore, we consider low glucose concentrations not to have played an important role in the pathogenesis of UI in our study population. Pregnancy‐associated anaemia, thrombocytosis, leukocytosis and mature neutrophilia were common findings in otherwise healthy bitches diagnosed with different forms of dystocia.     

Comparison of Intrauterine Antibiotics versus Ozone Medical Use in Sheep with Retained Placenta and Following Obstetric Assistance

This study outlines a new approach to reproductive tract treatment using ozone foam spray for certain ovine obstetrical problems, such as retained foetal membranes and possible uterine infections following obstetric assistance (OA), in comparison with classical antibiotics treatments. The study was conducted on 256 ewes from 11 sheep farms in north‐western Croatia. A total of 139 ewes were diagnosed with dystocia (DT) and 49 with retention of placenta (RP). Ewes with RP were treated either with ozone foam spray (Riger spray G; Novagen^®) applied into the body of the uterus for 2–3 s (first or RPO group; n = 24) or with two foaming, intrauterine tablets of oxytetracycline hydrochloride (Geomycin^® F) (second or RPA group; n = 25). The third and fourth groups consisted of ewes that received OA for dystocia (including ringwomb, foetal oversize and assistance of abnormal position and posture). The third group (DTO; n = 70) was treated with ozone foam spray, while ewes in fourth group (DTA; n = 69) were treated with antibiotics. The ewes in the control group (CTL) with physiological puerperium were randomly selected (n = 70) from all herds. Transrectal ultrasonography (transversal diameter of uterine horns) was used for the control of uterus regression on days 2 and 25 after parturition. There was a difference in transversal uterine horn diameter in the RP groups, that is RPO and RPA (5.40 ± 0.53 cm vs. 5.43 ± 0.40 cm), ewes with dystocia, that is DTO and DTA (5.37 ± 0.49 cm vs. 5.54 ± 0.60 cm) and ewes from the CTL group (4.98 ± 0.35 cm) one day after parturition. Average transversal uterine diameter of all groups at day 25 post‐partum was 1.80 ± 0.15 cm. The intrauterine ozone treatment in ewes with RP and after manual obstetrics attained similar results to spontaneously delivered ewes (CTL group), showed as the physiological regression of the uterus with a similar transversal diameter without the presence of lochia in the uterine lumen, indicating that this could be a novel potential alternative therapy.     

Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.     

“Subnutrition effects during pregnancy and lactation on mitosis,apoptosis and androgen receptor expression in the rat testis”

In order to study the effect of undernutrition during gestation on the testicular development in rats and its impact on mitosis, apoptosis and the relative abundance of androgen receptor expression, twenty primiparous 3‐month‐old Sprague–Dawley (Rattus norvegicus) rats, weighing 246 ± 4.0 grams when the experiment began, were mated by the same male. Control group (CG), n = 10, fed ad libitum with water and rat chow and restricted group (RG, n = 10) fed throughout pregnancy and until birth with 40% of the ad libitum maternal daily feed intake. Litters from both groups suckled for 25 days, RG with 14 pups/litter and CG with 8 pups/litter. After weaning, all animals had access to ad libitum food and water. Testicular samples were taken from male pups at 2, 25 and 100 days of age. Immunohistochemistry was used to evidence androgen receptor (AR) expression in apoptotic (caspase 3‐positive) and proliferating (PCNA‐positive) cells. Three hundred nuclei of sustentacular (or Sertoli: SC), interstitial (or Leydig: LC), myoid (MC) cells as well as gonocytes (GC) were evaluated. Neither LC nor GC showed any differences between groups. However, SC androgen receptor (AR) positivity index in neonatal animals was lower in RG (1.27 ± 0.22 vs. 1.65 ± 0.17**). MC showed lower AR positivity index at 2 (2.69 ± 0.046 vs. 2.8 ± 0.055**) and 25 (1.34 ± 0.097 vs. 1.56 ± 0.1***) days of life; at 100 days of life, there was a greater number of apoptotic MC in the RG (8.5 ± 0.4 vs. 2.95 ± 1.1***). Thus, the present experiment demonstrates that the population dynamics of MC are affected by foetal programming due to undernutrition.     

Serum concentrations of leptin in pregnant and non-pregnant bitches

Leptin regulates body weight and several physiological processes including reproduction. We evaluated the circulating levels of leptin in pregnant and non-pregnant bitches as well as their correlation with body weight, food intake and number of foetuses. Nineteen healthy German shepherd bitches were used and divided in two groups (pregnant n = 12 and non-pregnant n = 7). Blood samples were collected every 15 days starting from ovulation (Day 0) throughout pregnancy (pregnant group, P) or throughout luteal phase (non-pregnant group, NP) In pregnant bitches, leptin concentrations increased from the day of ovulation (1.32 ± 0.06 ng/ml) up to day 45 (1.51 ± 0.06 ng/ml; p < .01) and returned to baseline values from day 60 post-ovulation. In non-pregnant bitches, leptin concentrations remained constant throughout the whole observation period (estimated marginal mean ± SE=1.33 ± 0.38 ng/ml). Pairwise comparisons showed significant differences between P and NP at day 45 post-ovulation (p < .05). Multivariable models indicated that, controlling for time and litter size, there was a positive relationship between leptin concentration and BW (p < .05) although Pearson coefficients showed that the correlation between BW and leptin was only significant in NP animals at day 45 (r = 0.76, p < .05). The multivariable approach also suggested that, holding BW and time constant, leptin concentrations tend to increase as the number of puppies increased (p = .06). Our study supports indirectly the contribution of the feto-placental unit to the circulating maternal leptin concentration.     

Anti‐Muellerian hormone concentration in bitches with histopathologically diagnosed ovarian tumours and cysts

Increased concentrations of Anti‐Muellerian hormone (AMH) can indicate a granulosa cell tumour as shown in women, mares and cows. To investigate AMH to differentiate canine granulosa cell tumour from other ovarian pathologies, we evaluated the ovaries of 63 bitches. Blood serum samples were collected before surgery for AMH analysis. Ovaries were submitted for histopathological examination. Fourteen bitches showed normal ovaries. These bitches had AMH values between 0.12 and 0.99 ng/ml. In 20 bitches ovarian cysts i.e., follicular cysts (n = 8), corpora lutea cysts (n = 7), subsurface cysts (n = 5) were diagnosed. These dogs had AMH values of 0.11–2.09 ng/ml. Bitches with small luteinized follicular cysts had slightly higher AMH values than those without ovarian alteration. In 29 cases ovarian neoplasms i.e., granulosa cell tumour (n = 9), epithelial tumours (n = 16), dysgerminomas (n = 3) and one sarcoma were identified. Anti‐Muellerian hormone values of bitches with an ovarian neoplasm except granulosa cell tumour ranged from 0.18 to 1.18 ng/ml. The AMH values of bitches with granulosa cell tumour ranged from 1.12 to ≤23 ng/ml and were significantly higher (p < .05) than in all of the other bitches. The cut‐off of 0.99 ng/ml gave a sensitivity of 100% and a specificity of 94.44% to diagnose granulosa cell tumour. In conclusion, markedly elevated AMH concentrations in bitches are indicative for a granulosa cell tumour. However, negative testing does not rule out the existence of small one. Differentiation of GCT from luteinized follicular cysts may especially be difficult.     

Quantitative ultrasound elastography and biometry of the bitch uterus in the early puerperium after vaginal delivery and caesarean section

Elastography is an actual imaging method used to qualitatively and quantitatively evaluate the elastic properties of tissues. The aim of this study was to evaluate and compare uterine tissue biometry and stiffness during post-partum period in brachycephalic bitches (n = 12) after c-section (GCS; n = 8) or normal delivery (GNB; n = 4). These animals were evaluated daily by abdominal ultrasound from the day of delivery until the 10th day post-partum; measuring uterine diameter, myometrial and endometrial thickness (mm) and shear wave velocity (SWV; m/s), by B-mode and ARFI (acoustic radiation force impulse) elastography, respectively. Uterine diameter was higher (p = .012) in animals submitted to c-section (15.26 ± 4.73 mm) than in normal birth (12.53 ± 2.64 mm) during the first 7 days post-partum. Uterine thickness gradually involuted in both groups (p < .0001), the myometrium during the first 9 days (p = .005) and the endometrium during the first 6 days (p = .003). The myometrial and endometrial SWVs were similar between types of delivery (p = .7846 and .8273) and presented a gradual increase (p = .411; .0043, respectively), during the first 10 days post-partum. It was concluded that bitches with normal delivery had smaller uterine thickness and faster puerperal involution than submitted to c-section, while uterine tissue stiffness was similar between delivery types and increased gradually during post-partum.     

Effect of aglepristone (RU534) administration during follicular phase on progesterone,estradiol-17β and LH serum concentrations in bitches

Aglepristone was administered in bitches during the follicular phase to evaluate its effects on progesterone, estradiol-17β and LH serum concentrations. Ten German Shepherds were divided into two groups (treated n = 5; control n = 5). Treated bitches received 10 mg/kg BW of aglepristone subcutaneously during the early follicular phase, 24 hr after and then 7 days later. The control group was injected, at the same time periods, with saline solution (0.3 ml/kg BW). For the steroid evaluations, blood was collected daily from the onset of proestrus until the first day of cytological dioestrus. For LH base-line serum determination, blood was also collected every 20 min for 2 hr at the onset of proestrus. For LH surge identification, blood was collected daily (every 6 hr) starting from the day of the first administration of aglepristone or saline solution until the first day of dioestrus. All animals ovulated but the treated group presented longer ovulation-dioestrus intervals than the control group (5.2 ± 2.2 days p < .05). Serum concentrations of the evaluated hormones were similar between experimental animals except for serum LH. Indeed, no LH peaks were detected in the treated group while LH surges were clearly observed in the control group (9 ± 1 days after the beginning of proestrus. In particular, the area under the curve for LH was significantly lower in treated than control animals (12 ± 4 ng/ml x Day; p = .01). In conclusion, administrations of aglepristone during the follicular phase of the bitch does not affect the steroid hormone patterns but does prevent the occurrence of a LH surge. This work raises significant questions and opens perspectives concerning the mechanisms of ovulation in bitches.     

Does serum anti‐Müllerian hormone levels always discriminate presence of the ovaries in adult bitches? Comparison of two ELISA kits

Anti‐Müllerian hormone (AMH) is produced in the ovary, and thus, it is an excellent marker of follicle pool in females. Current interest is the clinical use of this parameter as a biomarker to assess presence or absence of an intact ovary and to diagnose ovarian remnant syndrome (ORS) following incomplete ovariohysterectomy (OHE) in bitches. The aim of this study was to evaluate serum AMH concentrations in bitches (n = 34) before and after OHE using two different commercial ELISA kits, one of which is based on detecting human AMH and the other is based on detecting human AMH and the other specified for canine AMH. Furthermore, serum AMH levels were also measured in six ORS cases to compare the diagnostic utility of the two different ELISA kits. Serum AMH concentrations measured using the human and canine kit prior to and after OHE were 0.32 ± 0.24, 0.006 ± 0.22 ng/ml (p < .001) and 12.08 ± 22.81, 9.55 ± 15.42 ng/ml (p = .868), respectively. Thus, the canine‐based kit was not able to reveal the significant drop in serum AMH levels. In conclusion, the human‐based ELISA kits successfully detected the drop in serum AMH concentrations. Reliable results can only be achieved from well‐designed ELISA kits, and AMH levels might be a useful diagnostic tool for the evaluation of presence or absence of ovaries as well as for the detection of ORS cases in bitches.     

Ultrasound during mid‐gestation: Agreement with physical foetal and placental measurements and use in predicting gestational age in sheep

To determine the effects of poor maternal nutrition and litter size on foetal growth during mid‐gestation, pregnant ewes (n = 82) were fed 100%, 60% or 140% of NRC TDN beginning at day 30.2 ± 0.2 of gestation. Transabdominal ultrasound was performed weekly between day 46.0 ± 0.4 and 86.0 ± 0.7 to monitor foetal heart width (HW), umbilical diameter (UMB), rib width (RW) and placentome outer (OD) and inner diameter (ID). Data were analysed with repeated‐measures using the mixed procedure for effects of maternal diet, litter size and gestation, and equations predictive of gestational age were generated using the regression procedure. To determine the agreement of ultrasound measurement and actual size, ewes (n = 20–21) were euthanized at day 45 or 90 to obtain corresponding postmortem measurements for Bland–Altman analysis. The HW, UMB and placentome OD and ID increased with gestation (p < .0001) but were unaffected by maternal diet or litter size (p ≥ .12). Ultrasound underestimated postmortem measurements of HW (14.8%), UMB (7.3%), placentome OD (4.5%) and ID (37.3%) at day 90 of gestation. Ultrasound underestimated RW at day 45 (7.7%) but overestimated RW (23.8%) at day 90, indicating inconsistent bias when reporting RW by ultrasound. Combining the HW, UMB, RW and placentome OD generated the strongest equation predictive of gestational age (R² = .91). These findings indicate that during mid‐gestation, maternal diet or litter size did not affect HW, UMB or placentome diameters and these factors can be used to estimate gestational age.     

Evaluation of a commercial intravaginal thermometer to predict calving in a Hungarian Holstein-Friesian dairy farm

In this study, the utility of a commercial intravaginal thermometer was evaluated as an automated method for the prediction of calving in a total of 257 healthy pregnant Holstein–Friesian female cattle. The accuracy and the sensitivity of predicting calving within 48 hr before calving were also evaluated. The intravaginal temperature changes from 72 hr before and up to calving were significantly (p ≤ .001) affected by parity, season (summer vs. autumn), the time of day (8 a.m. or 8 p.m.) and the 6-hr time intervals (38.19°C: first interval 0 to 6 hr before calving vs. 38.78°C: twelfth interval 66 to 72 hr before calving), while the gender (p = .943), and the weight of the calf (p = .610), twinning (p = .300), gestation length (p = .186), foetal presentation (p = .123), dystocia (p = .197) and retention of foetal membranes (p = .253) did not affect it significantly. The sensitivity of the SMS of expecting calving within 48 hr and the positive predictive value were 62.4% and 75%, respectively, while the sensitivity and the positive predictive value for the SMS of expulsion reached 100%. It can be concluded that the investigated thermometer is not able to predict calving within 48 hr accurately; however, imminent calving can be accurately alerted.     

Molecular studies on oestrogen α and progesterone receptors and histomorphometric analysis of canine uteri following aglepristone treatment

Aglepristone, a competitive progesterone antagonist, is successfully used in various progesterone-dependent conditions. This study investigated uterine histomorphometric analysis, and expressions of the oestrogen α receptor (ERα) and progesterone receptor (PR) in uteri of bitches following the single dose of aglepristone treatment. Twelve client-owned healthy diestrous bitches were used in the study. The single dose of aglepristone (Alizine^®, 10 mg/kg) was injected subcutaneously 5 days before ovariohysterectomy in the treatment group (n = 6); bitches without treatment served as a control group (n = 6). Uteri were collected for histomorphometric analysis, ERα and PR gene, and protein expressions studies. The mRNA expressions of ERα and PR were determined by RT-qPCR. Immunohistochemical analysis was used to evaluate the ERα and PR protein expressions using an H-score in five parts of the uterus. The results demonstrated glandular epithelium height significantly decreased (p < .05) and ERα mRNA increased (p < .01) in treated dogs. Of the treated bitches, lower expression levels of ERα were observed in the luminal epithelium, crypt and glandular epithelium, with higher expression in the endometrial stroma and myometrium (p < .05); however, PR expression decreased in the luminal epithelium, crypt and glandular epithelium (p < .01). In conclusion, reduction of the uterine glandular epithelium and ERα mRNA upregulation together with changes in ERα and PR expressions were observed in the treated bitches. However, changes in uterine ERα and PR expressions in the treated bitches depended on tissue layers. The treatment had no effect on serum oestradiol and progesterone levels.     

Factors associated with ewe and lamb survival,and subsequent reproductive performance of sheep undergoing emergency caesarean section

Clinical records of all 212 ewes undergoing emergency caesarean surgery at a veterinary teaching hospital between January 2008 and December 2019 were evaluated retrospectively. Their age ranged from 1 to 10 years (median = 4 years), with German merino the predominant breed (48.1% of cases). The most frequently diagnosed indications were insufficient cervical dilatation (n = 94, 44.3%), uterine torsion (n = 50, 23.6%), foetopelvic disproportion (n = 31, 14.6%) and vaginal prolapse intra partum (n = 11, 5.2%). Fifty-four (25.5%) of the 212 ewes additionally suffered from one or more concurrent, pre-existing conditions. Overall ewe mortality until hospital discharge was 10.8% (23/212), and 3.8% (n = 6) for the 158 ewes without a history of concurrent disorders. Mortality during hospitalization increased to 31.5% (17/54) for those with pre-existing conditions. Total lamb mortality was 49.1% (173/352) until hospital discharge. Pre-existing conditions (p = .001) and the presence of post-surgical complications (p = .025) were identified as significant factors influencing dam mortality, while delayed presentation for veterinary attention with an observed duration of labour of >12 hr was identified as the most influential factor on total lamb mortality (p = .010). The presence of dead or emphysematous foetuses was not significant for ewe mortality. Follow-up information on further outcomes was available for 156 (82.5%) of the 189 discharged ewes. Eighty-nine animals (57.1%) were re-bred in the following season and achieved a 93.3% (83/89) pregnancy rate, while the remainder had either been slaughtered (n = 56, 35.9%), sold (n = 5, 3.2%) or had died of unknown causes (n = 3, 1.9%). The subsequent incidence of dystocia was 15.6% (n = 12) in the 77 ewes with available information on lambing ease. Adequate management of underlying conditions and timely intervention are important factors for best possible short-term outcomes. In the long term, the subsequent pregnancy rate was good and the incidence of subsequent dystocia was within the normal range.     

Growth of foetal bones and metabolic profile during gestation in primiparous ewes and multiparous ewes

Primiparous ewes and multiparous ewes show physiological differences during pregnancy, which can have an impact on the development of their offspring. The objective of this study was to compare the changes in the metabolic profile and in the size of some foetal bones throughout gestation between primiparous and multiparous ewes. Twelve primiparous (PM) ewes and 14 multiparous (MT) ewes were used. According to the dates of lambing, two groups of ewes were formed: Group 1 (G1, n = 6 PM and n = 7 MT) and Group 2 (G2, n = 6 PM and n = 7 MT). The body weight, body condition score, metabolic and foetal morphometric parameters were determined from before conception until the end of gestation. After lambing, the body weight and survival rate during the first 72 hr of life of lambs, as well as the maternal behaviour score were recorded. The PM ewes were lighter (p < .01) and had a greater mobilization of body reserves during gestation, mainly evidenced by a greater serum concentration of NEFAs and lower serum concentration of total proteins (p < .05) compared with the MT ewes. The parity did not affect the foetal morphometric variables. The lambs of MT ewes were heavier at parturition (p = .002) and tended to have a greater survival rate than those lambs of PM ewes (p = .09). In conclusion, PM ewes and MT ewes differ in their metabolic profile throughout the gestation. However, in the present study, we did not find parity differences in the dimensions of foetal bones during growth in gestation.     

Some observations on the aerobic bacterial flora of the genital tract of the dog and bitch

The aerobic bacterial flora from the genital tracts of 143 bitches and 51 dogs was investigated. Beta haemolytic streptococci (group G), Staphylococcus aureus and Escherichia coli were among the organisms isolated from normal bitches in heat, abnormal bitches and normal dogs. Repeated sampling of two bitches during oestrus, and the dog to which they were mated, showed that bacteria were transmitted from bitch to dog, but did not become established. Guarded swab samples taken from the vagina collected less bacteria than samples from the vestibule. Numerous bacteria were seen in vaginal smears taken from bitches during oestrus, but numbers fell rapidly with the appearance of leucocytes in the smear at the onset of metoestrus.     

Intestinal development of bovine foetuses during gestation is affected by foetal sex and maternal nutrition

We aimed to evaluate the effects of maternal nutrition (MN) and foetal sex on the intestinal development of bovine foetuses throughout different days of gestation (DG). Forty‐four multiparous, dry Holstein × Gyr cows with average initial body weight of 480 ± 10 kg were fed the same diet of either restricted feeding at 1.15% of body weight (CO, n = 24) or fed ad libitum (overnourished, ON, n = 20). Six cows from CO group and five cows from ON group were slaughtered at 139, 199, 241 and 268 DG, and foetuses were necropsied to evaluate the intestinal development. The mass, length and density of foetal intestines were not affected by MN (p ≥ 0.260). An interaction between MN and DG was observed for the villi length of jejunum (p = 0.006) and ileum (p < 0.001). Villi length of jejunum and ileum was higher (p < 0.10) in foetuses from ON‐fed cows than in foetuses from CO‐fed cows at 139 DG. However, at 199 DG, the villi length of jejunum and ileum of foetuses from CO‐fed cows was higher than in foetuses from ON‐fed cows. Despite these differences, MN did not affect the villi length of jejunum and ileum at 268 DG (p > 0.10). Female foetuses had greater small intestine mass (p = 0.093), large intestine mass (p = 0.022), small intestine mass in proportion to body mass (p = 0.017) and large intestine mass in proportion to body mass (p < 0.001) than male foetuses. Female foetuses had also longer small intestine (p = 0.077) and greater small intestine density (p = 0.021) and villi length of jejunum (p = 0.001) and ileum (p = 0.010) than males. We conclude that MN affects the pathway for the development of foetal villi length throughout the gestation in bovine foetuses without changing the final villi length. Female foetuses had higher intestinal mass, density and villi length than males during the foetal phase in bovines.     

Bacteriological Status of Canine Milk and Septicaemia in Neonatal Puppies – a Retrospective Study

Results of the bacteriological examination of milk samples from 46 bitches were evaluated retrospectively, and correlated with findings of the bacteriological examination of organs from dead, septicaemic puppies in their litters (n = 33). The aim of this study was to investigate, in how many cases of clinical and subclinical mastitis of the bitches, the same bacteria can be detected in their septicaemic neonates. One group of lactating bitches was clinically healthy (group I, n = 38), whereas in eight bitches different puerperal disorders were found (group II). Twenty‐five septicaemic puppies were from group I, eight from group II. Out of a broad spectrum of bacteria isolated from the milk of clinically healthy and diseased bitches, only Escherichia coli, Klebsiella pneumoniae and/or ß‐haemolytic (haem.) Streptococcus sp. could be isolated from organs of their septicaemic puppies. This was the case in three bitches with mastitis and in one clinically healthy bitch only. Staphylococcus intermedius, although frequently isolated from canine milk, does not seem to be a cause of septicaemia in neonates. It is assumed that in most cases of neonate septicaemia, bacteria from the bitches’ milk are not the primary cause.     

Activity of selected glycosidases and availability of their substrates in bovine placenta during pregnancy and parturition with and without retained foetal membranes

The activity of glycosidases is crucial for the function and biological activity of proteins conjugated with sugar moieties, which play an important role in adhesion of cells during attachment and detachment of the foetal membranes. The aim of study was to describe the ability of bovine placental tissues to break down O-glycosidic bonds in different glycoproteins by the determination of activity of β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase and sialidase in early–mid-pregnancy as well as at parturition with released and retained foetal membranes. Moreover, the availability of substrates for these glycosidases in placental homogenates was evaluated. Placental samples were collected from pregnant (2–4 months) cows in slaughterhouse (n = 8) as well as during Caesarean section and divided into released foetal membranes (n = 8) and retained foetal membranes (n = 8). Tissue homogenates were subjected to spectrofluorimetric and spectrophotometric determinations of enzyme activities as well as electrophoretic separations. Enzyme activities expressed changes within examined time with significant (p < .05) differences between pregnancy and physiological parturition in β-N-acetyl-hexosaminidase and α-l-fucosidase in foetal part of placenta while in maternal part only in the latter one. Decreasing tendency in enzyme activity was noticed in foetal part of retained samples in comparison with released ones with significant (p < .05) differences in α-l-fucosidase activity. The analysis of staining of sugar moieties attached to selected proteins depicted availability of sugar molecules in examined tissues, but their patterns differed between samples. In conclusion, sugar moieties in conjugated proteins express changes in the course of pregnancy which is reflected by the alterations in activities of placental glycosidases.     

Tissue distribution of Leishmania chagasi and lesions in transplacentally infected fetuses from symptomatic and asymptomatic naturally infected bitches

Visceral leishmaniasis (VL) is primarily transmitted by an invertebrate vector, but transmission in the absence of the vector has been reported. Vertical transmission of VL has been described in man and dogs. The aim of this study was to evaluate the distribution of Leishmania amastigotes in fetal organs and histopathologic changes associated with parasitism and to determinate the frequency of transplacental transmission and potential of vertical transmission by symptomatic and asymptomatic pregnant bitches. Symptomatic (n = 4) and asymptomatic (n = 4) pregnant bitches, serologically and parasitologically positive for Leishmania sp., carrying a total of 53 fetuses (26 from symptomatic and 27 from asymptomatic bitches) were selected at the Veterinary Hospital of the National University of Asuncion, Paraguay. Samples of placenta and fetal organs such as liver, spleen, lymph nodes, bone marrow, kidney and heart were histologically evaluated and processed for immunodetection of amastigotes and PCR. There were no lesions compatible with VL in fetal tissues in spite of the presence of amastigotes, particularly in lymphoreticular tissues. However, fetal hepatocytes had marked degenerative changes that were independent of the presence of amastigotes in liver. Twenty-six out of 53 placentas (13 symptomatic and 13 asymptomatic) and a total of 17 fetuses out of 53 (nine symptomatic and eight asymptomatic) were PCR positive. Together these findings indicate a high frequency of transplacental transmission and no differences in the potential of transmission when symptomatic were compared to asymptomatic pregnant bitches.