Determination of gestational time and prediction of parturition in dogs and cats: an update.

Accurate prediction of delivery date in canine and feline allows a better management of parturition, reducing the loss of neonates. This review evaluates the most common methods adopted to accurately predict the day of delivery: determination of ovulation and hormonal assays, first appearance of embryonic/foetal structures using ultrasound or radiography, echographic measurement of extra-foetal and foetal structures, or evaluation of foetal flux and heart rate. Determination of ovulation and hormonal assays at the time of breeding and close to pregnancy term is widely used to predict parturition in dogs (Concannon et al. American Journal of Veterinary Research 44, 1983, 1819; Hayer et al. Journal of Reproduction and Fertility, Suppl. 47, 1993, 93; Hase et al. Journal of Veterinary Medical Science, 62, 2000, 243; Kutzler et al. Theriogenology, 60, 2003a, 1187). In cats, some studies have been carried out, but no hormonal parameters for accurate prediction of parturition have been described so far (Buff et al. Journal of Reproduction and Fertility, Suppl. 57, 2001, 187; De Haas van Dorsser et al. Biology of Reproduction, 74, 2006, 1090; DiGangi et al. Journal of the American Veterinary Medical Association, 237, 2010, 1267; Dehnhard et al. Theriogenology, 77, 2012, 1088). Many studies suggested that gestational timing can be obtained by observation using ultrasound or radiography of specific structures in relation to the time of appearance during gestation (Concannon and Rendano American Journal of Veterinary Research, 44, 1983, 1506; Rendano et al. Veterinary Radiology, 25, 1984, 132; Shille and Gontarek Journal of the American Veterinary Medical Association, 187, 1985, 1021; Davidson et al. Veterinary Radiology, 27, 1986, 109; England et al. Journal of Small Animal Practice, 31, 1990, 324; Yeager et al. American Journal of Veterinary Research, 53, 1992, 342; Zambelli et al. Theriogenology, 57, 2002a, 1981; Zambelli et al. Journal of Feline Medicine and Surgery, 4, 2002b, 95; Zambelli and Prati 2006; Lopate Theriogenology, 70, 2008, 397; Davidson and Baker Topics in Companion Animal Medicine, 24, 2009, 55). Ultrasonographic measurement of extra-foetal and foetal structures is a common and accurate method for the prediction of parturition day during pregnancy, when specific formulae are used depending on the ultrasonographic parameter, the species and, in canines, the size of the bitch (Shille and Gontarek Journal of the American Veterinary Medical Association, 187, 1985, 1021; England et al. Journal of Small Animal Practice, 31, 1990, 324; Luvoni and Grioni Journal of Small Animal Practice, 41, 2000, 292; Luvoni and Beccaglia Reproduction in Domestic Animals, 41, 2006, 27; Lopate Theriogenology, 70, 2008, 397; Michel et al. Reproduction in Domestic Animals, 46, 2011, 926; Beccaglia and Luvoni Reproduction in Domestic Animals, 47, 194, 2012). Recent studies demonstrated that in dogs, the imminence of parturition could be predicted by evaluating foetal flux and foetal heart rate by ultrasound (Gil et al. Theriogenology, 82, 2014, 933; Giannico et al., Animal Reproduction Science, 154, 2015, 105). For an accurate prediction of parturition date, the combination of different methods is desirable.

Pharmacokinetics and perioperative efficacy of intravenous ketorolac in dogs.

Ketorolac (KET) is a nonsteroidal anti-inflammatory drug approved for the use in humans that possesses a potent analgesic activity, comparable to morphine, and could represent a useful tool to control acute pain also in animals. The clinical efficacy and pharmacokinetic profile of intravenous (IV) ketorolac tromethamine (0.5 mg/kg) were studied in 15 dogs undergoing gonadectomy. Intra-operative cardiorespiratory variables were monitored, and post-operative pain was assessed using a subjective pain score (0-24) in all dogs, whereas the pharmacokinetic profile of the drug was determined in 10 animals. During surgery, mean minimal alveolar concentration of isoflurane was 1.69 ± 0.11%, and normocapnia and spontaneous ventilation were maintained in all animals. During pain assessment, no significant differences between males and females were found, and in no case rescue analgesia was necessary. No adverse effects were reported. Serum samples were purified by solid-phase extraction and analysed by HPLC with UV-Vis detection. A large variability was observed in serum concentrations. The kinetics of ketorolac was described by a noncompartmental analysis. The elimination half-life (t½λz ) and ClB were 10.95 ± 7.06 h and 92.66 ± 84.49 mL/h/kg, respectively, and Vdss and Vz were 1030.09 ± 620.50 mL/kg and 1512.25 ± 799.13 mL/kg, respectively. AUC(0→last) and MRT(0→last) were 6.08 ± 3.28 h × µg/mL and 5.59 ± 2.12 h, respectively. The results indicate that ketorolac possess good post-operative analgesic effects until about 6 h after administration in dogs undergoing moderately painful surgery.

Prediction of parturition in dogs and cats: accuracy at different gestational ages.

In bitches and queens, the ultrasonographic measurement of extrafoetal and foetal structures allows the evaluation of gestational age and the prediction of the parturition term for an extended period of time. The aim of this study was to investigate whether the accuracy of parturition date prediction is affected by the week of pregnancy when the ultrasonographic examination is performed. The results were obtained by retrospective analysis on the gestational period basis (from week 4 to week 9 of pregnancy) in 495 ultrasonographic examinations of pregnant bitches (small and medium size) and 60 of pregnant queens. They demonstrated that a similar accuracy (p > 0.05) was obtained by the measurement of inner chorionic cavity at week 4 and 5 of pregnancy (± 1 day, 81% vs 67.7%; ± 2 days, 93.1% vs 85.9%). Accuracy (± 1 day) based on biparietal (BP) measurement was similar at week 5 and 6 of pregnancy (78.6% vs 78.9%; p > 0.05), whereas a significant decrease (p < 0.05) was observed at week 7 and 8 compared to week 6. No difference in the accuracy of the prediction at ± 2 days based on BP was observed from week 5 to 8 of gestation (range from 82.3% to 95.2%). Close to term (week 9), the accuracies of the prediction based on BP were significantly lower than those obtained at week 5 (± 1 day: 50.9% vs 78.6%, p < 0.05; ± 2 days: 69.8% vs 95.2%, p < 0.005). The accuracy of the prediction in bitches and queens is influenced by the gestational period when ultrasonographic measurements are performed, particularly for BP measurement. Although a decrease in BP accuracy occurs along the pregnancy, the accuracy at ± 1 and ± 2 days of prediction of parturition term is highly consistent for 6 and 8 weeks of gestation, respectively.

Are Italian-Polish veterinarians and breeders prepared to control an outbreak of Brucella canis infection in dogs?

Brucella canis infection is one of the most important causes of infertility in dogs and is a zoonosis for which no effective treatment or vaccines exist. It is not a mandatory notifiable disease. Following an increase of cases in Europe and worldwide, an investigation was performed to evaluate how much Italian and Polish veterinarians and breeders know about canine brucellosis and understand their perceptions of this infection. For this reason, two questionnaires were prepared, in Italian and Polish. Eighteen Italian and Polish veterinarians, specialists in canine reproduction, responded to the first survey and 44.4% of them affirmed having diagnosed canine brucellosis at least once in their clinical practice, and different perceptions emerged regarding the infection in the two countries. The second survey was completed by 145 Italian and Polish breeders; the disease was completely unknown to 22.8% of them, whereas 2.1% had diagnosed infection by B. canis in their kennels. In conclusion, knowledge of B. canis infection differs between these countries, with extremes ranging from diagnosed cases to complete underestimation of the presence of the problem. However, based on international data and reporting of a recent large outbreak in Italy, awareness of this contagious infectious disease and its management must be increased.

TRIS-lecithin extender supplemented with antioxidant catalase for chilling of canine semen.

The aims were to evaluate the suitability of a non-commercial TRIS-lecithin (LC) extender and the effect of different concentrations of catalase (CAT) on motility, capacitation status (Chlortetracycline-assay) and zona pellucida (ZP) binding capacity of canine spermatozoa stored at +5 degrees C for 4 days. The sperm-rich fractions of the ejaculates of four stud dogs were divided into four aliquots. After centrifugation, sperm pellets were diluted (200 x 10(6) sperm/ml) in TRIS buffer, citric acid, fructose, antibiotics, supplemented with 20% egg yolk (TRIS-EY) or 0.04% soybean lecithin (TRIS-LC) with CAT (150 or 450 UI/ml) or without CAT, and then preserved at 5 degrees C for 4 days. The results showed that LC is a valid alternative to EY for chilling canine semen, as similar rates of motility, number of uncapacitated spermatozoa and of spermatozoa binding the oocyte ZP were obtained in semen chilled in TRIS-LC or TRIS-EY. Different concentrations of CAT in a TRIS-LC based extender did not improve the quality of semen after chilling. However, a concentration of 150 UI/ml CAT resulted in an increased number of spermatozoa bound to the oocyte ZP, after 4 days of chilling when compared to semen chilled with TRIS-EY and TRIS-LC. In conclusion, an animal protein-free extender with soybean LC, as a replacement of EY, is suitable for 4 days chilling of canine spermatozoa, but the addition of CAT does not improve general semen quality except for a slight effect on sperm-ZP binding.

Isolation of canine mammary cells with stem cell properties and tumour-initiating potential.

Recent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self-renewal or by progenitor cells that have acquired the capacity for cell self-renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long-term non-adherent culture conditions. We showed that cells obtained from spheres that display self-renewing properties, have multi-lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour-initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.

Ultrasonographic study of deep portion of diencephalo-telencephalic vesicle for the determination of gestational age of the canine foetus.

The aim of this study was to evaluate the accuracy of ultrasonographic measurement of the deep portion of diencephalo-telencephalic vesicle (DPTV) in the prediction of delivery date in different size bitches. The effects of litter size and foetal sex ratio on the accuracy were also investigated. For this purpose, the growth curve of DPTV was derived in large size dogs (26-40 kg) and the results of the accuracy of the prediction were compared with those obtained in small (< or =10 kg) and medium (11-25 kg) size dogs by the application of the equations derived from the growth curve previously described. Ultrasonographic examinations were performed once a week during the second half of pregnancy in seven large size bitches (26-40 kg body weight). A linear regression model was adopted to analyse the relationship between the DPTV mean values and the days remaining to parturition. The results of regression analysis indicated that DPTV measurement in large size dogs is significantly and linearly related to the gestational age. Ultrasonographic measurements of DPTV were also performed during pregnancy in different size bitches with unknown breeding dates. Although the results indicated a similar accuracy of the prediction of the date of parturition in the different sizes of bitches, a higher accuracy was obtained in normal and large litter size compared with small litters. Foetal sex ratio did not affect the accuracy. In conclusion, this study demonstrated that the accuracy of the prediction of parturition day obtained by ultrasonographic evaluation of DPTV growth is reliable when specific formulae for different size dogs are applied. It should be noted that although foetal sex ratio does not affect the accuracy, the prediction could be less accurate when a small litter is present.

Unusual vaginal angiomatous neoformation in a 3-year old pug.

A 3-year-old female pug (8 kg bodyweight) was referred by a local veterinarian for evaluation of vaginal bleeding. Historically, the owner indicated chronic haematic vaginal discharge initiated approximately 3 months ago, overall good appetite and slight general dullness. Two months prior to the visit, the bitch had been spayed by the local veterinarian, but vaginal bleeding persisted after surgery. Following a general examination and complete blood analysis, a genital tract examination was performed and, through vaginal endoscopy, a red, smooth, and apparently broad-based vaginal mass was found. An episiotomy was performed and on surgical exploration, the bleeding site was determined to be on the surface of a mass composed of two congested, adjacent cylindrical structures, symmetric to the vaginal sagittal midline, broad-based and quite regular on the surface. The mass appeared to be well delimited from the surrounding tissue by a thin capsula, and each cylindrical structure had a tributary vessel. Ectopic and dysplastic corpora cavernosa of the clitoris characterized by angiomatous proliferation, cavernous haemangioma and vascular hamartoma were considered in the differential diagnosis. The presence of small intralesional nerves identified by S100 immunostaining was used as a diagnostic clue to classify this unusual vaginal angiomatous neoformation as vascular hamartoma.

Spontaneous reduction of intracranial arachnoid cysts: a complete review.

The aetiopathology, clinical features and treatment of arachnoid cysts (AC) are still extremely controversial topics. The posterior fossa is the second most common site of these lesions, since they are often detected in the cerebellar or cerebellar-cistern region. Despite this, almost all the cases of non-surgical reduction of an AC reported in literature concern cysts of the middle fossa, whereas only two cases of spontaneous reduction of a posterior fossa AC has been described in literature. A complete review of the literature regarding this topic has been collected and discussed. The authors present a case of spontaneous reduction of an arachnoid cyst situated along the midline of the posterior cranial fossa with regression of all neurological symptoms. This 43-year-old man presented increasing nuchal headache with vomiting and nausea, simulating subarachnoid haemorrhage. CT and MRI documented an arachnoid cyst along the midline of the posterior fossa compressing the mesencephalon. The patient did not present any family history of this pathology nor had suffered head trauma. The patient was only given analgesics to relieve painful symptoms. After 30 days, spontaneous resolution of all the neurological symptoms took place. A 2 months, MRI control documented marked reduction of the cyst without any evident brain compression. The feasibility of a wait and see policy in such cases is discussed.

[Imaging of acute brain inflammatory disease]. / Imaging della patologia infiammatoria acuta dell'encefalo.

The central nervous system inflammatory disease can be due to any kind of infective agent (bacterial viral, fungal and parasitic), but entails also multiple sclerosis, a primary demyelinating disease in which the causal agent is unknown. MR imaging is, in most often, the procedure of choice, due to her multiplanar and multiparametric imaging, and to her better contrast resolution. The post-contrast imaging with double dose of gadolinium and late sequences enable visualisation of smallest pathologic foci or slightest blood-brain barrier alterations, with a sensibility very higher than post-contrast CT scan. In addition, RM provide to many functional informations, by means of diffusion, perfusion and spectroscopy studies, Bold technique for cortical activation studies and Fiber Tracking technique, in order to demonstrate pathologic modification earlier than they are evident on morphologic imaging. Functional imaging is also employed to monitor response to treatment and damage reversibility.

Comparison of the accuracy of two ultrasonographic measurements in predicting the parturition date in the bitch.

OBJECTIVES: To compare the accuracy of two ultrasonographic measurements, inner chorionic cavity diameter and biparietal diameter, for the prediction of parturition date in small and medium size bitches. The effects of litter size and fetal sex ratio on the accuracy of the prediction were also investigated. METHODS: Ultrasonographic examinations were performed on pregnant bitches and, depending on the stage of pregnancy, the inner chorionic cavity diameter and biparietal diameter were measured. The mean value of the data of each structure was calculated and the predicted day of parturition was obtained by the application of equations derived from growth curves we previously developed for different size bitches. RESULTS: The parturition term was predicted with similar accuracy during early gestation, by the measurement of inner chorionic cavity diameter, as well as during late gestation, by the measurement of biparietal diameter. Litter size affected only biparietal diameter accuracy, whereas fetal sex ratio did not influence the estimation of the day of parturition. CLINICAL SIGNIFICANCE: Inner chorionic cavity diameter and biparietal diameter were equally reliable parameters in obtaining a reasonably accurate prediction of parturition date at any gestational period at which a bitch is examined.

Prediction of whelping date in large and giant canine breeds by ultrasonography foetal biometry.

OBJECTIVES: To derive the growth curves of the inner chorionic cavity and the biparietal diameter in large and giant dog breeds by ultrasonographic foetal biometry. To evaluate their accuracy in the prediction of whelping date and the effect of litter size and foetal sex ratio. METHODS: Foetal biometry parameters were obtained using serial ultrasonographic examinations in eight large (26 to 40 kg) and nine giant (>40 kg) pregnant bitches with known breeding dates and concentrations of serum progesterone during oestrus. The relationship between inner chorionic cavity or biparietal diameter growth and days to parturition were analysed by linear regression and the equations derived from the growth curves were applied to predict the whelping day. The accuracy of the prediction (whelping day ±1 day and ±2 days) and the litter size and sex ratio were recorded. RESULTS: The results showed a significant relationship between days before parturition and inner chorionic cavity or biparietal diameter. The overall accuracy at ±2 days was greater, than that at ±1 day. In giant breed bitches, the accuracy of the prediction by biparietal diameter was significantly lower in small, than normal litter size. No effect of foetal sex ratio was observed. CLINICAL SIGNIFICANCE: Foetal biometry parameters obtained by ultrasonography can be used to predict whelping dates in large and giant dog breeds.

Pharmacokinetics, intraoperative effect and postoperative analgesia of tramadol in cats.

Tramadol is a synthetic codeine analogue used as an analgesic in human and veterinary medicine, but not approved for use in cats. Tramadol (2 mg/kg) was administered intravenously (IV) as preoperative analgesic in 12 cats (6 males) undergoing surgical gonadectomy. The pharmacokinetic profile of the drug and its O-desmethyl metabolite were determined in 8 animals (4 males), while intraoperative effects and postoperative analgesia, estimated by subjective pain score (0-24), were evaluated in all. Mean intraoperative isoflurane consumption was reduced, but hypoventilation was not observed. Sex-related differences were not observed, particularly in terms of postoperative analgesia: rescue analgesic was never administered. Concentrations of the active O-desmethyl metabolite were persistently high in all the animals. Considering the results obtained in this study, tramadol, at the dose of 2 mg/kg IV, did not produce any evident intraoperative cardiorespiratory side effects and with additional investigation may prove to be an appropriate intraoperative analgesic in cats undergoing gonadectomy.

The prediction of parturition date in canine pregnancy.

In bitches, the length of gestation is highly variable when measured from the day of mating, thus prediction of parturition may be greatly inaccurate when determined from this point. The ability to precisely predict the duration of pregnancy is of practical importance for managing parturition or planning caesarean section and this review analyses the methods that can be used to accurately forecast the day of parturition. At the time of breeding, determination of the luteinizing hormone surge and the initial rise in progesterone provide reliable information on gestational length of the bitch, but when pregnancy is ascertained, ultrasonography is the most useful tool to predict the delivery day. In fact, by ultrasonographic measurements of the extra-foetal and foetal structures an accurate prediction can be made both in early and late pregnancy.

Embryo production in dogs: from in vitro fertilization to cloning.

Increased availability of canine embryos would be desirable to develop research and to apply assisted reproductive technologies in the treatment of infertility and in the improvement of reproductive performances in valuable Canids, both domestic and non-domestic. Embryo production through in vitro fertilization and nuclear transfer has been technically achieved in the dog, and the transfer of cloned embryos has recently resulted in the birth of puppies. However, the efficiency of these technologies is still very limited. This is mainly because of the peculiar characteristics of the canine oocyte and the lack of its full acquisition of developmental competence in vitro. This paper discusses the latest results and aspects on which further research should be focused to provide advances in the field.