Deslorelin subcutaneous implants in Oryx dammah males for reproductive control

The aim of this study was to assess the effects of the deslorelin subcutaneous implant as a temporary contraceptive method in the Oryx dammah male. For this purpose, deslorelin at different doses, i.e. 14.1 mg and 9.4 mg, was subcutaneously implanted in three males (Phase 1) and one male (Phase 2) adult Oryx dammah, respectively. Quantitative behavior evaluation and androgen concentrations in feces and plasma were assessed before and after implant application. Fecal androgen concentrations observed in treated males were compared with those measured in one orchiectomized male and two females. Fecal androgen concentrations increased up to 15 days after the implant application, then progressively decreased, reaching the basal level at day150 in Phase 1. In Phase 2, levels remained high until day 60 and returned to basal level on day 120. Plasma testosterone concentration was higher on the day of implant application than three months later, but with variable ranges among males. A general increase of activity levels and hierarchical changes were observed after treatment, in accordance with hormonal variations. Despite males cohabiting with two fertile females during the observation period, no births were recorded. However, between the end of Phase 1 and the beginning of Phase 2, i.e. about 10–11 months after the first deslorelin implant, a fertile mating occurred leading to the birth of a calf. Therefore, we can hypothesize a contraceptive effect up to 10 months after the implant. Testicular histology performed on one male at the end of the Phase 2 showed no spermatogenetic activity. Our results suggest that deslorelin implant can be used to temporarily control reproduction in the Oryx dammah male. Behavior and fecal androgen measurements were useful and repeatable, non-invasive methods to monitor response

Fetal propofol and dexmedetomidine exposure during elective c-section in the bitch: impact on pup viability

Elective C-section is a common procedure recommended in dogs at risk of dystocia(1). Anesthetics administered at surgery can cross the placenta leading to distress up to neonatal mortality(2). The study aims to determine the impact on pup viability of a new anesthetic-analgesic protocol for elective C-section in the bitch. For this purpose, propofol (PPF) and dexmedetomidine (DEX) concentrations in maternal blood, amniotic fluid and placental tissue were correlate to neonatal parameters. Nine purebred bitches (age 4,9±2,3 years; weight 39,8±10,4 kg) were induced with a combination of PPF at 2,5 mg/kg and DEX at 2 μg/kg IV and maintained with isoflurane in 100% oxygen for elective C-section. If needed, additional doses of PPF were administered to effect in order to achieve intubation of patients. DEX and PPF quantification from the different biological matrices was carried out by HPLC-MS and HPLC-FL methods. Neonatal viability at birth was assessed with a modified Apgar score(3) (AS) and birth weight recorded as well as mortality of pups within 48 hours of life. Results: A total of 54 pups was delivered, 77,80% of them recorded as vigorous by AS. Neonatal mortality was 11,1%. Lowest AS was assigned to pups from mothers receiving additional dose of PPF. AS was not influenced by birth weight of pups, nor by maternal and placental drugs concentrations. Maternal blood PPF (range 0,24-2,8- mcg/mL) and DEX (range 0,41-2,04 ng/mL), and placental PPF (range 0,24-2,57 mcg/mL) concentrations tended to decrease over time, while placental DEX (range 1,32-6,15 ng/mL) was fairly uniformly detected in pups from the same litter. DEX concentration in placenta was much higher than in maternal blood showing a greater placental retention compared with PPF. Both PPF and DEX were not detectable in amniotic fluid. Placenta resulted an effective barrier against fetal DEX exposure making this protocol safe, analgesic and advisable for elective C-section in dog. (1) Moon PF, Erb HN, Ludders JW, et al. Perioperative Risk Factors for Puppies Delivered by Cesarean Section in the United States and Canada. J Am Anim Hosp Assoc 2000;36:359-68. (2) Luna SPL, Cassu RN, Castro GB et al. Effects of four anaesthetic protocols on the neurological and cardiorespiratory variables of puppies born by caesarean section. Vet Rec 2004;154:387–89 (3) Groppetti D, Pecile A, Del Carro AP, et al. Evaluation of newborn canine viability by means of umbilical vein lactate measurement, apgar score and uterine tocodynamometry. Theriogenology 2010;74(7):1187-96

Treatment Effects and Risk Factors Evaluation in Longitudinal Studies : A Statistical Help for Data Analysis

This paper was inspired by the experience of the Authors research group composed by oncologist veterinarians and a biostatistician to evaluate treatments and prognostic factors with the aim to help veterinarians involved in longitudinal studies into evaluating and writing prognostic results. Longitudinal studies are commonly analysed by techniques for survival data, taking into account for the time elapsed from the beginning of observation and the occurrence of an event related to treatment effect or disease course. The presence of incomplete follow-up information for some subjects requires specific descriptive and inferential statistical methods. Two literature datasets were analysed to show statistical models implementation techniques and to discuss statistical issues: I) A multicentre clinical trial on remission maintenance of children with acute Lymphoblastic leukaemia and II) A randomized clinical trial on advanced inoperable lung cancer. Data sets concerned studies on \u201chumans\u201d, nevertheless the peculiar data structure allowed to discuss some aspects which are common to survival analysis studies, regardless on subject\u2019s characteristics. Log-rank test was used to compare survival curves for treatments and the relationship between Log-Rank test and univariate Cox model results was explained. As the evaluation of prognostic impact cannot be based only on p-values, the strength of the association between treatments and prognosis was estimated to take into account for the clinical relevance of results. On the second data set, beside of treatment, other clinical variables were available and a multivariate Cox model was applied. Model implementation was discussed concerning the coding of categorical variables and the relationship between continuous variables and model response. Suggested rules for the maximum number of covariates to be included in order to obtain reliable results were cited. Finally, the predictive ability of the model was discussed based on a measure of the area under ROC curve specific for survival data

Immunohistochemical insights into a hidden pathology : Canine cryptorchidism

Cryptorchidism is a common disorder in the canine population with some aspects still unclear. Although the bilateral condition is known to lead to fertility problems and predisposition to testicular cancer, the neoplastic risk for scrotal testis in unilateral cryptorchid dog is controversial. Therefore, the therapeutic approach to the canine unilateral cryptorchid is arbitrary so far. This study aimed to investigate precancerous testicular lesions, such as immaturity and atrophy, and compare them in scrotal and undescended testes using an in-depth diagnostic analysis based on immunophenotypic patterns. With this purpose, 26 adult male dogs of different ages and breeds, affected by unilateral or bilateral cryptorchidism were enrolled. After surgical removal, testes were examined immunohistochemically to assess their positivity for specific markers of the canine foetal/neonatal period, that is vimentin (VIM), cytokeratin (CK), desmin (DES), inhibin-α (INH), and anti-Müllerian hormone (AMH) in Sertoli cells, and placental alkaline phosphatase (PLAP) in germ cells. Except for the ubiquitous VIM, all the markers were more expressed in neoplastic gonads compared to healthy ones (P < 0.05). Similarly, testes detected with Sertoli cell-only tubules as well as with Sertoli cells hyperplasia showed higher expression than gonads without such alterations for CK, DES, AMH and PLAP, and for CK and DES, respectively (P < 0.05). The same trend was observed in undescended respect to scrotal testes even though their positivity was significant only for DES, INH and AMH (P < 0.05). Immunohistochemical positivity found in scrotal testes of unilateral cryptorchid dogs, even in absence of detectable anatomical abnormalities, was suggestive of precancerous lesions. Despite the limited sample size, this study could help to clarify the predisposition to neoplastic development in normally descended testes. These markers expression in adult life could allow identifying the early stages of the testicular carcinogenesis process besides suggesting a precautionary bilateral surgical approach in unilateral cryptorchid dogs

Can peri-surgical electroacupuncture relieve immunity suppression? A pilot study in dogs

General anesthesia and surgical stress can suppress the immunological response by acting both directly on the immune system and indirectly on the hypothalamic-pituitary-adrenal axis and the sympathetic nervous system. Disturbance of the immune system during the perioperative period can lead to complications such as wound-healing disorders and infections up to sepsis. Effectiveness of acupuncture in regulating the immune function by increasing leukocyte numbers and inhibiting inflammatory response has been proven. This study aimed to explore the impact of electroacupuncture (EAP) on the dynamic balance of the immune system and immune cell populations in dogs undergoing surgery. Twelve healthy bitches scheduled for elective ovariectomy were divided into two groups according to whether (EAP, n=6) or not (CTR, n=6) a peri-operative electroacupuncture treatment was performed. Levels of leukocytes (neutrophils, monocytes, T- and B-cells) and immunoglobulins M (IgM) and A (IgA) were measured in blood samples collected before (T0), 1 hour (T1) and 2.5 hours (T2) after anesthesia induction. Leukocytes count decreased from T0 to T1 in both groups and restored within 1.5 hours in EAP group whereas remained significantly lower in CTR group (P<0.02). In particular, neutrophils and monocytes increased in dogs receiving EAP (P<0.01) while T-cells decreased in CTR group (P<0.04) at T2. B-cells and cytotoxic T-cells decreased in EAP dogs (P<0.04) at T2. No differences in helper T-cells, IgM and IgA levels were recorded between groups and over time. Our results suggest a modulatory effect of EAP on the immune system which is early expressed on neutrophils, monocytes and T-cells

Neonatal mortality in dogs : prognostic value of Doppler ductus venosus waveform evaluation - Preliminary results

Aim: To define the prognostic value of Doppler ultrasonographic morphology of ductus venosus (DV) waveform on canine neonatal mortality. Materials and Methods: Fifty-four healthy pregnant bitches underwent fetal ultrasonographic assessment. The DV waveforms were classified as diphasic (dDVw) or triphasic (tDVw) and compared with neonatal mortality. Results: Ninety-three fetuses were evaluated. Twenty fetuses belonged to litters with neonatal mortality, in which tDVw was observed. Seven fetuses belonged to litters without neonatal mortality, in which tDVw was observed. Fifty-eight fetuses belonged to litters without neonatal mortality, in which only dDVw was observed. Eight fetuses belonged to litters with neonatal mortality, in which only dDVw was observed. The correlation between tDVw and neonatal mortality was statistically significant (odds ratio [OR], 20.7; p<0.0001). Considering only pregnancies with one or two fetuses with the same DV waveform: Two fetuses with tDVw belonged to litters with neonatal mortality; 1 foetus with tDVw belonged to litter without neonatal mortality and 26 fetuses showed dDVw without neonatal mortality. The correlation between tDVw and neonatal mortality even in litters up to two pups was statistically significant (OR, 88.3; p=0.01). Conclusion: Echo-Doppler assessment of DV is feasible in canine fetuses, and the presence tDVw seems to be related to neonatal mortality

Characterization of a population of unique granular lymphocytes in a bitch deciduoma, using a panel of histo- and immunohistochemical markers

The ovaries and uterus were collected after ovariohysterectomy from a 16-month-old Labrador bitch in diestrus that never mated. Discrete swellings were found in the uterine horns, with the macroscopic appearance of normal early pregnancy. At histologic examination, the endometrium, devoid of any conceptus and chorion, showed a marked proliferation, on the basis of which a diagnosis of deciduoma was made. A remarkable population of stromal eosinophilic granular lymphocytes was present, especially in the axis of the endometrial folds. Periodic acid–Schiff and Dolichos biflorus–lectin histochemical reaction and a panel of 10 immunohistochemical markers were used to characterize eosinophilic granular cells. Our findings allowed us to compare these granular cells with the granulated decidual cells, whose presence was until now described only in primates, rodents, or a few other epitheliochorial species. On the basis of our results, the importance of eosinophilic granular cells in a decidualization process is hypothesized to occur also in the bitch

Persistent M&#252;llerian Duct Syndrome in a German Shepherd Dog

In mammals, the regression of the m\ufcllerian ducts is regulated by the action of the AMH hormone which is produced by testes during embryonic development. The action of this hormone is mediated by the only known receptor AMHR2. Mutations occurring in the AHM hormone and/or in the AMHR2 receptor gene cause the lack of regression of m\ufcllerian ducts, which may therefore persist even in male embryos carrying a XY chromosomal arrangement. This is known as the persistent m\ufcllerian duct syndrome (PMDS). A female German Shepherd dog was referred to the veterinary clinic because of urinary incontinence. She also showed an anatomical structure that protruded from and enlarged the vulvar labia. From the morphological appearance, one gonad resembled an ovary and the other a testicle. The histological examination instead showed that the gonads were both testes with an underdeveloped parenchyma and without signs of spermatogenetic activity. No alterations were found with regard to the uterus which showed a correctly developed body, cervix, and horns. Genetic analysis, performed on DNA extracted from blood, showed (i) the presence of both X and Y chromosomes, (ii) the absence of chromosome XX/XY chimerism, (iii) a normal SRY gene coding sequence, (iv) a normal AMHR2 gene coding sequence, and (v) a normal AMH gene coding sequence. In this study, we report and characterize a new case of PMDS in a dog excluding that the only mutation hitherto found in the AMHR2 gene is responsible for the observed phenotype

Asymptomatic unilateral ovarian leiomioma in a German shepherd bitch

This report shows for the first time clinical imaging (ultrasound and computed tomography), histological and immunohistochemical findings of an ovarian leiomyoma, coincidentally diagnosed in an asymptomatic unmated nulliparous ten year-old German shepherd bitch concurrently suffering from multiple mammary tumors. A thorough examination allowed the differentiation of ovarian leiomyoma from other spindle cell tumors. An accurate description of the diagnostic procedures useful in the managing of ovarian leiomyoma could provide valuable information to veterinary practitioners. Indeed, despite its rarity and nonspecific symptoms, ovarian leiomyoma may also affect the dog with an unknown potential risk

A first glance on the epigenome of Capra hircus

DNA methylation and microRNAs (miRNA) are two important forms of epigenetic modifications that play an important role in gene regulation in animals. Methylation at the carbon 5 position of cytosine residues is a fundamental layer of cellular differentiation through the control of transcriptional potential. MiRNA are small noncoding RNA molecules that regulate gene expression. Complete DNA methylomes for several organisms are now available; at the present, methylome of the domestic goat is unexplored. There is also still limited knowledge about miRNAs expression profiles in small ruminant species. Therefore, to contribute information on epigenetic modification in Capra hircus, we analysed the methylome and the miRNA population of three tissues (hypothalamus, pituitary and ovary) from 3 adult Saanen goats. We used Methylated DNA binding domain sequencing with enrichment of methylated DNA fragments and next generation sequencing. We produced least 23 million reads per sample, which were aligned to the goat reference genome. Further analyses were performed to identify peaks corresponding to hyper-methylated regions. We sequenced miRNAs expressed in the three tissues with Illumina high-throughput sequencing. Reads were mapped on the Capra hircus reference genome and both known and novel miRNAs, and miRNA target sites were identified using information collected in miRBase and using specific bioinformatic tools. This study produced a comprehensive miRNA profile related to the biology of goat. Furthermore, this is the first work dealing with methylome in Capra hircus: our preliminary results could provide new information for a deeper comprehension of epigenetic mechanisms of this species