Insights into alternative cryoprotectants to freeze sperm of domestic cats.

Globalization and habitat destruction pose a significant threat to wildlife felids. Even though conservation banks for genetic materials have been created, the sperm cryopreservation with minimal cell damage is still a great challenge. Thus, this study aimed to compare the effects of two commercial extenders with different concentrations of alternative cryoprotectants on thawed sperm quality of domestic cats. Five adult cats were anaesthetized (using a combination of 40 µg/kg medetomidine associated to 5 mg/kg ketamine), and the semen was collected by electroejaculation (electrical stimulation of 2-3 V). Semen samples were evaluated for sperm characteristics (kinetics, morphology, membrane integrity and morphometry). Subsequently, they were sorted into two aliquots and centrifuged. The aliquots were added to a commercial extender containing 3% glycerol and 2% methylformamide (extender I) or 2% glycerol and 3% methylformamide (extender II), frozen, thawed (37°C/30 s) and reevaluated. Comparatively, the sperm kinetics and membrane integrity of fresh semen were higher (p < .002) than frozen samples in extender I and II. Total and progressive motility were lowest in the thawed samples. However, the subjective analysis indicated high sperm motility, since the kinetics evaluation was impaired by the low cell number in the thawed samples. There were no differences in sperm morphology between the groups. In the sperm morphometric analysis, a significant difference (p = .04) was identified in the length of the intermediate piece in extender II samples compared with fresh and extender I. Thus, it can be concluded that although the concentrations tested did not maintain the kinetic parameters and membrane integrity of spermatozoa after thawing, the extender with a lower concentration of glycerol was less toxic for maintaining the midpiece length.

Renal biomarkers, clinical parameters, and renal Doppler velocimetry in bitches with cystic endometrial hyperplasia-pyometra complex.

Cystic endometrial hyperplasia (CEH)-pyometra complex is the most common uterine infection in adult and elderly bitches and can cause renal dysfunction. The aim of this study was to measure and compare urinary creatinine, urea, symmetric dimethylarginine (SDMA), urinary protein-creatinine ratio (UPC), measurement of systolic blood pressure (SBP), and Doppler velocimetry of renal arteries in patients with CEH-pyometra complex before and after an average of 6 months of treatment, evaluating the possibility of the changes persisting. The evaluation was conducted at two moments: M1 (at the diagnosis of CEH-pyometra, n = 36) and M2 (after an average of six months of treatment, n = 16). For the control group, eight bitches with no changes in blood tests or history of conditions underwent Doppler ultrasound evaluation of the renal arteries. At both M1 and M2, we measured creatinine, urea, and serum SDMA, UPC, SBP, and Doppler ultrasound of the renal arteries. Patients were evaluated according to the following groups: azotemic (AZO) and non-azotemic (NAZO), and open and closed cervix pyometra. The parameters were compared between animals present in both moments presented as M1R (bitches that were in M1 and M2) and M2. Statistical significance was considered when p < .05. The medians found for creatinine in M1 were as follows: 1.15 mg/dL, being 1.8 mg/dL for AZO (12/36) and 0.95 mg/dL for NAZO (24/36); and in M2: 0.85 mg/dL (16/16), being 1.15 mg/dL for AZO (4/16) and 0.8 mg/dL for NAZO (12/36). For urea, in M1 it was 36 mg/dL (32/36), with AZO being 103 mg/dL (11/32) and 33 mg/dL in NAZO (21/32); and in M2 32 mg/dL (16/ 16), being 29 mg/dL for AZO (4/36), and 31 mg/dL for NAZO (3/15). The median SDMA measured in M1R was 17 µg/dL (15/16), with AZO being 31 µg/dL (3/15), and NAZO being 16.5 µg/dL (12/15); and in M2, SDMA was 12 µg/dL (16/16), with AZO being 12.5 µg/dL (4/16), and NAZO being 12 µg/dL (12/16). The median UPC measured in M1 was 1.15 (10/36), with AZO being 0.25 (1/10), and NAZO being 1.38 (9/10); and in M2, it was 0.2 (13/16), being 0.1 in AZO (4/13), and 0.2 (9/16) in NAZO. For SBP, in M1, it was 118 mmHg (30/36), with AZO being 102 mmHg (10/30) and 133 mmHg in NAZO (20/30); and in M2 142.5 mmHg (12/16), being 155 mmHg for AZO (4/12), and 140 mg/dL for NAZO (8/12). When comparing animals with open and closed cervixes, a difference was found between SDMA measurements (p = .001). There was a distinction between PI and RI of the left and right kidneys consecutively (p = .007; p = .033; p = .019; p = .041). Correlations found in M1: SDMA × PI RIM DIR (r = 0.873; p = .002), SDMA × PSV RIM ESQ (r = 0.840; p = .004), SDMA × EDV RIM ESQ (r = 0.675; p = .046). With this study, we conclude a return to normality of renal biomarkers and clinical parameters after six months. Yet, there is a persistence of Doppler velocimetric measurements between the two moments. Thus, this parameter is not suitable for identifying and classifying chronic kidney injury in bitches with pyometra.

Emergency cesarean section in dogs: Usefulness of amniotic fluid biochemical parameters and placental morphology as indicators of neonatal viability.

In recent years, special attention has been paid to the analysis of fetal fluids and placental histopathology to identify parameters that can be used as indicators of maternal reproductive quality, embryonic viability, and fetal and neonatal health. Newborn health reflects the functioning of the fetal adnexa and its relationship with maternal tissues. Therefore, evaluating these components is promising for the early detection of newborns at risk. This study aimed to detect the biochemical characteristics of the amniotic fluid (AF) and histopathological characteristics of the placenta for comparison between canine neonates born by elective (EL) and emergency (EM) cesarean sections (CSs) and associate the results with neonatal viability in the first 24 h. A total of 38 neonates born by ELCS (n = 19) and EMCS (n = 19) were selected. AF was collected to analyze the concentration of its biochemical components [alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatinine, urea, total protein, albumin, total and direct bilirubin, lactate, glucose, potassium, chloride, calcium, and sodium]. Histopathological processing of the placenta was used to describe the lesions and identify the arrangement of collagen fibers using hematoxylin-eosin and picrosirius staining. There was an increase in ALP activity (P = 0.035) and the concentrations of lactate (P < 0.001) and potassium (P = 0.031), and a decrease in chlorides (P < 0.001) in the AF of neonates in the EMCS group. The comparisons between the groups did not show differences between the presence and extent of lesions in the placenta; however, a difference was observed in the arrangement of collagen fibers in the placental structure. A comparison between AF and histopathological findings showed a negative correlation (r = -0.609, P = 0.003) between glucose concentration and the presence of necrosis in the placental labyrinth. It was observed that the composition of the AF changed owing to the influence of the type of cesarean, possibly caused by prolonged hypoxia in cases of dystocia. ALP activity and lactate, potassium, and chloride concentrations in the AF might be explored as markers of neonatal health in EMCS. Under the conditions of this study, no correlations were found between the placenta's histopathological characteristics and the neonates' viability.

Comparison of two pharmacological semen collection times with α2-adrenergic agonist in domestic cats.

The use of α2-adrenergic agonists in association with urethral catheterization has been used as a technique for pharmacological semen collection in cats. The mechanism of action of this drug is the stimulation of adrenoreceptors in the vas deferens, which results in ejaculation. While medetomidine is the α2-agonist most commonly used in studies, ejaculation with the use of dexmedetomidine associated with ketamine has been effective, but with variable results. Therefore, further studies regarding the methodology of use are required to obtain better seminal quality. This study aimed to compare two pharmacological semen collection times after the association of dexmedetomidine (30 µg/kg, IM; Dormitor®, Zoetis), ketamine (5 mg/kg, IM; ketamine, Vetnil) and urethral catheterization using a tomcat probe (0.8 mm × 1.00 mm × 11 cm). The collections were divided into two experimental groups: G10 (N = 8; urethral catheterization after 10 min of anaesthesia) and G15 (N = 8; urethral catheterization after 15 min of anaesthesia). The ejaculates were evaluated for ejaculate volume, sperm concentration, morphology and kinetics using the CASA system. To compare the groups, the t-test and the Mann-Whitney U-test were used with a significance level of 5%. It was identified that ejaculate volume (G10: 22.62 ± 2.13 vs. G15: 26.81 ± 1.55; p < .001) and sperm concentration (G10: 48.10 × 106 ± 17.84 vs. G15: 90.18 × 106 ± 19.35; p < .001) was higher in G15 than in G10 and had a lower percentage of minor defects than G10 (G10: 3.12 ± 2.41 vs. G15: 1.00 ± 1.19; p = .043). Regarding the kinetic parameters, the results of G15 were better for total motility-TM (G10: 67.00 ± 10.33 vs. G15: 81.87 ± 7.99; p = .006) and faster cells-RAPID: (G10: 55.00 ± 16.63 vs. G15: 74.25 ± 11.94; p = .019); whereas a higher proportion of cells with slow speed-SLOW were seen in G10 (G10: 31.00 ± 12.07 vs. 17.12 ± 7.53; p = .015). Based on these findings, we suggest that collection via urethral catheterization should be performed 15 min after the application of ketamine-associated dexmedetomidine to obtain a better-quality ejaculate.

Fecal microbiota profile of ovariohysterectomized cats submitted to estrogen replacement / Perfil da microbiota fecal de gatas ovariohisterectomizadas submetidas à reposição estrogênica

This study aimed to observe the effects of 17 ß-estradiol replacements on the fecal microbiota in spayed cats. Individual samples of fresh feces were collected and stored at -80° C. Sequencing of the V3/V4 regions of the 16S rRNA gene was used, and bioinformatic analysis was performed. Firmicutes/Bacteriodetes ratio was lower in the group receiving estrogen replacement compared to the SHAM group (P = 0,005). Jaccard index (P = 0.123) and Yue & Clayton index (P = 0.094) did not reveal alpha and beta diversity differences. The linear discriminant analysis effect size (LefSe) identified Firmicutes and MegasPhaera as the biomarkers for the SHAM group, and Burkholderiales, Betaproteobacteria, Sutterellaceae, Suterella, Proteobacteria, Proteobacteria unclassified and Collinsella for the group receiving estrogen replacement.(AU)

O objetivo deste estudo foi observar os efeitos da reposição de 17 ß-estradiol na microbiota fecal de gatas castradas. Amostras individuais de fezes frescas foram colhidas e armazenadas a -80°C. Foi realizado o sequenciamento das regiões V3/V4 do gene 16S rRNA e a análise bioinformática. A razão Firmicutes/Bacteriodetes foi menor no grupo que recebeu reposição estrogênica em comparação ao grupo SHAM (P = 0,005). O índice de Jaccard (P = 0,123) e o índice de Yue & Clayton (P = 0,094) não revelaram diferenças na alfa e beta diversidade. A análise discriminatória linear de tamanho do efeito (LefSe) identificou Firmicutes e Megasphaera como biomarcadores para o grupo SHAM, e Burkholderiales, Betaproteobacteria, Sutterellaceae, Suterella, Proteobacteria, Proteobacteria não classificada e Collinsella para o grupo que recebeu reposição estrogênica.(AU)

The proAKAP4 concentrations in Nelore bull sperm and their relation to FTAI conception rate results.

The AKAP4 protein has an essential role in sperm motility, and its precursor, the proAKAP4, is considered a biomarker for sperm quality and fertility. Despite this, proAKAP4 concentration was never evaluated in Bos indicus sperm. Therefore, this study aims to determine the proAKAP4 concentration in the sperm of Nellore bulls and its association with sperm kinetics, morpho-functionality, morphology, and conception rates after fixed-time artificial insemination (FTAI). The bulls (n = 9) used in our study presented sperm characteristics above expected standards for quality. Sperm from each bull was evaluated regarding kinetics by the CASA system, and assessed for mitochondrial functionality, sperm membrane integrity, and morphology. For the FTAI protocol, multiparous Nelore cows (n = 1507) were inseminated with frozen-thawed sperm from the same batches used in the laboratory analysis. Additionally, the sperm proAKAP4 concentration was determined using an ELISA kit. The data were analyzed by linear regression and the significance level was set to 5%. The mean conception rate was 52.52%. The mean proAKAP4 concentration was 25.75 ng/106 spermatozoa. The proAKAP4 concentration positively affected the sperm total and progressive motilities, and conception rates after FTAI (respectively, r2 = 0.49, P = 0.03; r2 = 0.63, P = 0.01; and r2 = 0.51, P = 0.03). Based on the relationship detected in this study, we conclude that the sperm proAKAP4 concentration is a suitable biomarker to predict the quality and fertility of frozen-thawed Bos indicus semen.

Sperm subpopulations influence the pregnancy rates in cattle.

This study aimed to study the characteristics and subpopulations of spermatozoa from bulls with low and high reproductive performance based on pregnancy rates. Based on historical records of pregnancy rate from four farms, 24 bulls were selected. Two groups were established, with low pregnancy rates (n = 12; LOW), including bulls that presented pregnancy rates <52.27% (33.33% to 51.81%); and a group with high pregnancy rates (n = 12; HIGH), with pregnancy rates >52.27% (52.27% to 69.64%), after fixed-time artificial insemination (FTAI). The thawed sperm straws were analysed to sperm kinetics, morphology, plasma membrane integrity and sperm subpopulations. The LOW group exhibited a higher proportion of static cells (p < .05). In contrast, the HIGH group showed greater percentages for membrane integrity and total and progressive motility, and cells with fast and medium velocity (p < .05). In the cluster procedures, four sperm subpopulations were established. The low-fertility bulls presented the highest percentage of subpopulation 2 (41.46%), characterized by slow and progressive spermatozoa. The high-fertility bulls exhibited the highest percentage of subpopulation 3 (37.17%), characterized by fast and nonlinear spermatozoa. Results from this study indicated that bulls with greater percentages of fast and nonlinear spermatozoa seem to have greater fertilization capacity and the subpopulations analysis can be considered a tool to identify ejaculates with high fertility.

Eletrocardiografia computadorizada em cadelas com tumores mamários / Electrocardiography in bitches with mammary tumors

The incidence of cancer has increased due to greater longevity of the animals as a consequence of better control of other diseases, improvement of nutrition and good practices in preventive medicine. Malignant tumors can cause paraneoplastic syndrome such as hypercalcemia, anemia, cachexia, among others and consequent cardiovascular disorders. The electrocardiography (ECG) is a complementary exam that can reveal with its traces these rhythmic disorders. Based on that, the objective of this study was to evaluate the ECG in bitches with mammary tumors in order to detect in which type of tumor would be found significant changes as in cardiac rhythm, ECG waves, segments or cardiac axis. Twenty-nine female dogs were used in this study, being 19 bitches with mammary tumor, and they were allocated in three groups: G1: control group (n = 10), G2: benign neoplasia group (n = 6) and G3: malignant neoplasia group (n = 13). The most prevalent type of neoplasia in G2 was the benign mixed tumor (83%), followed by adenoma (17%) whereas in G3: carcinoma in mixed tumor (61%); papillary carcinoma (23%). Regarding cardiac rhythm, it was found sinus arrhythmia (SA) and normal sinus rhythm (NS): G1: 50% SA and 50% NS; G2: 67% SA and 33% NS; G3: 54% SA and 46% NS. No ventricular or atrial arrhythmias were detected. For other parameters in G1, G2 and G3, respectively (mean ± Std error): FC (bpm): 110±9.2, 120 ± 8.5, 124±7,5; P (ms): 48 ± 1.6, 51 ± 1.8, 50 ± 1.2; P (mV) 0.19 ± 0.02, 0.2 ± 0.02, 0.19 ± 0.02; PR (ms): 94 ± 4.3, 93 ± 5.5, 89 ± 3.9; QRS (ms): 56 ± 1.54, 60 ± 4, 62 ± 1.2; R (mV): 1.1 ± 0.06, 1.2 ± 0.24, 0.9 ± 0.13; QT (ms): 203 ± 9.4; 204 ± 7.9; 182 ± 15.6; and cardiac axis (°): 66 ± 6.2, 61 ± 7.9, 70 ± 7.5. There were no significant differences for all cardiac parameters and also for ST interval and T wave morphology. All electrocardiographic parameters found are in accordance with other studies carried out in the canine species. The results regarding tumor types differ from what was found in another study, in which in 18 bitches with mammary tumors, 55% were benign, with prevalence of adenomas (38%) followed by benign mixed tumors, and 45% malignant, with adenocarcinoma prevailing (22%). In another study, it was verified in 63 bitches the predominance of tubular carcinoma (26.56%) and carcinoma in mixed tumors (23.44%). Diverging from the electrocardiographic parameters of this study, a significant difference was found in the R wave amplitude value in the research by Barros et al., (2015) who performed computerized electrocardiography in 50 dogs, not only with mammary neoplasms (55% mammary carcinomas), but also in mastocytomas, lymphomas, benign tumors and other sarcomas. This author found out that the R wave amplitude values of the neoplasia group were lower when compared to the control group. In addition, we suspect that other types of tumors could result in more paraneoplastic syndrome than the mammary neoplasms found in this research. Neoplasms as lymphomas (T cells), apocrine gland anal sac adenocarcinomas, multiple myelomas and thymomas are known as a cause of hypercalcemia of malignancy and histiocytic sarcomas, myelomas, leukemia and lymphomas causing anemia. In conclusion, ECG has no changes in benign or malignant mammary tumors in dogs. However, this fact does not exclude the importance of its performance in pre-anesthetic evaluations. Further studies with a larger sample including the clinical staging of these bitches with a balanced number of animals with low and high staging are suggested.(AU)

Influence of long-term thermal stress on the in vitro maturation on embryo development and Heat Shock Protein abundance in zebu cattle.

The objective of this study was to investigate the influence of long-term temperature stress during the in vitro maturation (IVM) of oocytes on the in vitro embryo production (IVP) and the abundance of HSP70 and HSP90 in zebu cattle. Viable cumulus-oocyte complexes (COCs) were incubated for 24 h at 37 °C, 38.5 °C, or 40 °C for the low-, physiological, and high-temperature stress treatments, respectively. Thereafter, they were subjected to in vitro fertilization and culture. Temperature did not affect the polar body extrusion. However, IVP was adversely affected when IVM took place at 37 °C and 40 °C. The highest abundance of HSP70 was observed in cumulus cells after maturation of COCs at 40 °C. In contrast, HSP70 was more abundant in oocytes at both 37 °C and 40 °C; however, at 40 °C, the difference to the control group (38.5 °C) was not significant. In contrast, the highest abundance of HSP90 was observed in oocytes and cumulus cells at 37 °C. It appears that HSP70 and HSP90 respond to cold and heat stress in different ways. In conclusion, moderately high (40 °C) and low (37 °C) thermal stress for 24 h during IVM is detrimental to the developmental competence of oocyte and is accompanied by changes in the abundances of HSP70 and HSP90, especially in cumulus cells.

Correlation of testicular artery Doppler velocimetry with kinetics and morphologic characteristics of epididymal sperm in dogs.

Sperm quality can be affected by a reduction in testicular blood flow, which can be measured by Doppler ultrasonography. The aim of this study was to correlate the Doppler velocimetry of the testicular artery with kinetics of the epididymal spermatozoa in dogs. Twenty-two dogs (44 testicles) were evaluated by Doppler ultrasonography in five regions of the testicular artery before orchiectomy. Spermatozoa were recovered by the epididymal tail compression technique and analysed for kinetics on a computer-assisted semen analysis (CASA system). Morphology (modified Karras) and sperm membrane integrity were analysed by eosin-nigrosine staining. Data were analysed by Pearson's correlation test (p < .01). The mean total motility was 69.0% ± 17.7, progressive motility was 43.7% ± 14.7, average path velocity (VAP) was 127.0 µm/s ± 20.7, curvilinear velocity (VCL) was 221.0 µm/s ± 31.1, and sperm velocity index (SVI) was 389.9 ± 56.1. There were positive correlations between the peak systolic velocity (PSV) in the proximal supratesticular region with the SVI (r = .529), VCL (r = .555) and VAP (r = .473), and a negative correlation with the percentage of slow spermatozoa (r = -.463). The results suggest that the testicular artery blood flow velocity can positively affect the speed of spermatozoa movement. For the first time, we have correlated sperm kinetics with the Doppler evaluation of the testicular artery in dogs.

Can testicular artery Doppler velocimetry values change according to the measured region in dogs?

Testicular tissue maintenance is performed by the testicular artery, and the hemodynamics of this artery can be evaluated using Doppler ultrasonography. The aim of this study was to characterize the peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) and resistivity index (RI) of five regions of the testicular artery in dogs, including two proposed regions and three that have been previously described. Twenty-two dogs were used, and the PSV, EDV, PI and RI of the testicular artery were measured in five regions: proximal, medial and distal supratesticular; marginal; and intratesticular. The median values for PSV (cm/s), EDV (cm/s), PI and RI in the five regions were as follows: proximal supratesticular (23.1, 3.7, 2.1 and 0.8); medial supratesticular (17.2, 4.5, 1.5, and 0.7); distal supratesticular (12.2, 5.7, 0.8, and 0.5); marginal (11.3, 6.5, 0.5, and 0.4); and intratesticular (5.7, 3.5, 0.5, and 0.4). There was a difference between the PSV of the medial and distal supratesticular regions. There were differences in the PSV, EDV, PI and RI among the distal supratesticular, marginal and intratesticular regions. Measurements of PSV, EDV, PI and RI of the testicular artery in dogs at the proposed regions showed different results due to the hemodynamic and morphological differences of the artery during its course in the spermatic cord and to the testicles. It is necessary to identify the region in testicular artery Doppler velocimetric evaluations of dogs, given that there is a difference according to the region measured.

Influence of cooling temperature in sperm subpopulations of domestic cats.

The objective of this study was to identify and compare domestic feline sperm subpopulations, chilled at -1 °C for 24 and 48 h, as well as to analyze the sperm frequency in different subpopulations. Ten adult cats were used. Sperm collection was performed using electroejaculation (EEJ). Spermatic kinetics were evaluated using a computerized system at three moments: fresh, 24 and 48 h after refrigeration. The ejaculates were divided into a group refrigerated at -1 °C (n = 5,) and a group refrigerated at 4 °C (n = 5),. A total of 1560 spermatozoa were analyzed individually, and the sperm subpopulations were identified using multivariate statistics. Three spermatic subpopulations were defined using prior analysis of the hierarchical dendrogram. A principal components analysis (PCA) identified the existence of three groups with higher iterations at the three moments: PC1 (VAP, VCL, VSL, ALH, SVI), PC2 (STR, LIN, WOB and SMI) and PC3 (BCF). Subpopulation 1, after 48 h of refrigeration at -1 °C, and subpopulation 3, after 24 h of refrigeration at 4 °C, maintained their sperm quality, which allowed us to characterize the groups of spermatozoa that were resistant to cryopreservation. The present study identified three well defined ejaculate spermatozoa subpopulations, with proportional distributions between the groups and two refrigeration resistant subpopulations.

Effects of chemical castration using 20% CaCl2 with 0.5% DMSO in tomcats: Evaluation of inflammatory reaction by infrared thermography and effectiveness of treatment.

This study used infrared thermography to monitor the immediate inflammatory reaction to an intratesticular injection of 20% (calcium chloride) CaCl2 with 0.5% dimethyl sulfoxide (DMSO) for chemical castration and to provide evidence of the treatment's clinical viability. Six animals received a 0.25 mL intratesticular injection of 20% CaCl2 with 0.5% DMSO in each testis. Thermographic imaging, testicular measurement, penile spine evaluation, electroejaculation, and sperm analysis were performed before the injection. Eighty days post-injection, the cats were evaluated again, then received an orchiectomy followed by histological analysis. Infrared thermography of the testicular area was performed before (M0) and after anesthesia (M0A); after electroejaculation (M0E); 10 min (M1), 1 h, (M2) and 6 h (M3) after the injection for seven consecutive days (M4-10); after 15 (M11) and 30 days (M12); and after 80 days, before and after anesthesia and after electroejaculation (M13, M13A and M13E). No cats had a significant increase in testicular temperature or behavioral changes. All animals were azoospermic and had a 50% reduction in total testicular volume after 80 days. Histologically, the testes showed different degrees of degeneration, necrosis, calcification and replaced connective tissue, as well as Leydig cell hyperplasia in 7/12 of the testes. Infrared thermography efficiently diagnosed and monitored scrotal inflammation caused by intratesticular injection of 20% CaCl2 solution with 0.5% DMSO, and it is suggested that a single injection of this compound can cause azoospermia after 80 days. We concluded by infrared thermography with a clinical exam, that the adverse reactions were minimal and did not interfere with animal welfare.

Cirurgia reconstrutiva com retalho cutâneo de avanço como técnica alternativa para tratamento de carcinoma de células escamosas em cães: relato de caso / Reconstructive surgery with skin flap advance as alternative technique for treating squamous cell carcinoma in dog: case report

O carcinoma de células escamosas (CCE) é uma das neoplasias mais frequentemente observadas no trato genital de cães. O tratamento de eleição para o CCE é a exérese com ampla margem cirúrgica (1 a 3 cm), porém quando a região prepucial está comprometida, se torna difícil obter margem adequada, tanto pela anatomia da região quanto pelas características agressivas dessa neoplasia. A penectomia com uretrostomia pré-escrotal seria a técnica mais indicada nesses casos, entretanto, complicações pós-cirúrgicas podem reduzir a qualidade e a expectativa de vida do animal. O objetivo deste relato foi demonstrar uma técnica cirúrgica conservativa, com base em plásticas reconstrutivas com retalhos cutâneos, como tratamento para CCE em prepúcio e região inguinal, associada ao uso do piroxicam como terapia adjuvante. O animal não apresentou recidiva do tumor ou complicações secundárias ao tratamento até a última avaliação, um ano após o término do tratamento.

Squamous cell carcinoma (SCC) is one of the most frequently neoplasms observed in genital tract in dogs. The treatment of choice for the CCE is the excision with wide surgical margin (1-3 cm), but when the foreskin region is committed becomes difficult to obtain adequate margin, due both the anatomy of the region and the aggressive characteristics of this neoplasm. The penectomy with pre-scrotal urethrostomy would be the most suitable technique in such cases, however, postoperative complications may reduce the quality and expectancy of life. The objective of this report was to present a conservative surgical technique, based on reconstructive plastic with skin flap as a treatment for SCC in prepuce and inguinal region, associated with the use of piroxicam as adjuvant therapy. The dog was evaluated during one year after the end of treatment and haven't showed any tumor recurrence or secondary complications to treatment.

Hemangiopericytoma in a female dog with direct invasion of abdominal cavity and pulmonary metastasis / Hemangiopericitoma em uma cadela com invasão direta da cavidade abdominal e metástase pulmonar

An intact adult female Poodle dog was presented with a history of an increase in volume in the left and right mammary inguinal glands. The histopathological examination revealed a proliferation of spindle cells arranged in bundles with concentric arrangements surrounding blood vessels, occasionally collapsed. Three weeks after the surgery, the animal presented a recurrence of the tumor that extended from the left inguinal mammary gland to the vulva. Necropsy revealed direct invasion of the abdominal cavity by the tumor and pulmonary metastasis. The definitive diagnosis of hemangiopericytoma was made by histopathological and immunohistochemical examination. Pulmonary metastasis of CHP (canine hemangiopericytoma) is rare and there is no previous report of direct invasion to the abdominal cavity as observed in the present case.

Uma cadela adulta, intacta, da raça Poodle, apresentou histórico de aumento de volume na mama inguinal direita. O exame histopatológico revelou proliferação de células fusiformes dispostas em bandas e arranjos concêntricos ao redor de vasos, os quais se apresentavam ocasionalmente colapsados. Três semanas após a cirurgia, o animal apresentou recorrência do tumor que se estendia até a vulva. No exame necroscópico, observou-se que a massa invadia a cavidade abdominal e presença de metástase no pulmão. O diagnóstico definitivo de hemangiopericitoma foi realizado por meio dos exames histopatológico e imuno-histoquímico. A metástase pulmonar do hemangiopericitoma canino é rara e não há relatos prévios de invasão direta para cavidade abdominal, como observado no presente caso.

The effect of season on serum testosterone concentrations in dogs.

Photoperiod and environmental temperature are two important factors that may influence the reproductive cycle of various species. The objective of this study was to investigate seasonal influences on serum testosterone concentrations in dogs in a tropical zone, where the variation in day length between winter and summer solstice was approximately 2.5 h. Blood samples were collected every 15 days from seven adult dogs over a 14-month interval and serum testosterone concentrations were determined by radioimmunoassay. The year was divided into four seasons and mean testosterone concentrations for each season were related to the mean environmental temperature and rainfall during that season. Mean testosterone concentrations were 1.81 ng/mL (winter 2002), 1.93 ng/mL (spring 2002), 1.31 ng/mL (summer 2003), 2.02 ng/mL (autumn 2003) and 1.93 ng/mL (winter 2003). The temperature ranged from 10.2 to 32.8 degrees C and the rainfall from 33 to 476 mm. Serum testosterone concentrations were lower in summer 2003 than in both spring 2002 (P = 0.05) and autumn 2003 (P = 0.016). In a tropical zone, a combination of high temperature and substantial rainfall may have reduced serum testosterone concentrations in dogs.

Herapin-binding proteins of canine seminal plasma.

Heparin-binding proteins (HBP) from seminal plasma have been expected to participate in modulation of the acrosomal reaction, and have been correlated with fertility in some species. However, they have not been described in the dog. The aim of this study was to document the HBPs of canine seminal plasma. Six pooled samples of seminal plasma from three crossbred dogs were used. The HBPs were isolated by heparin affinity chromatography and the fractions recovered were pooled. One-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was carried out on 12 and 18% vertical minigels. The stained gels were scanned and the molecular weight (kDa) values for each band within a lane were calculated by image analysis software. The electrophoresis analysis of the pooled eluded fractions identified 19 bands, with molecular weights varying from 61.5 to 5.2 kDa. Previous studies, using one-dimensional SDS-PAGE, identified two bands (67 and 58.6 kDa), which were positively correlated with some semen parameters (sperm motility, sperm vigor, percentage of morphologically normal sperm and plasma membrane integrity). The 61.5 kDa band detected in the present study apparently corresponded to the 58.6 kDa band identified previously. Canine seminal plasma contained HBP; since HBP modulate the acrosome reaction in other species, they may have the same function in the dog. Further studies are necessary to better characterize this protein and determine if it is associated with fertility in the dog.

Vasectomy effect on canine seminal plasma biochemical components and their correlation with seminal parameters.

Three semen samples were collected at 48 h intervals from 20 mature research dogs previously conditioned to manual semen collection. Vasectomy was performed in all dogs, and 15 days after surgery, another three ejaculates were similarly collected. The semen was evaluated, and centrifuged to obtain seminal plasma for measurement of pH, and concentrations of total proteins (TP), total chlorides (Cl), calcium (Ca), potassium (K), and sodium (Na). The seminal plasma protein profile was evaluated by SDS-PAGE; molecular weights and the integrated optical density (IOD) of each band were estimated. There was a negative correlation between K concentration and progressive motility (r = -0.49, P = 0.027), sperm vigor (r = -0.60, P = 0.0053), and plasma integrity, evaluated by both the hypo-osmotic swelling test (r = -0.50, P = 0.026) and a fluorescent stain (r = -0.45, P = 0.046). Positive correlations between Na and K pre- and post-vasectomy (r = 0.88, P < 0.001; r = 0.56, P < 0.01, respectively) were verified. There were a total of 37 bands pre-vasectomy and 35 post-vasectomy (range, 100.6-3.6 kDa). Bands B9 and B13 (42.6 and 29.2 kDa) were not present post-vasectomy. The IOD of band B3 (73.5 kDa) was higher (P = 0.03) pre-vasectomy, compared to post-vasectomy; conversely, the IODs of bands B29 and B37 (7.8 and 3.6 kDa) increased (P = 0.026 and 0.047). Pre-vasectomy, there was a positive correlation (r = 0.49, P = 0.029) between band B37 band (3.6 kDa) and the Na:K ratio. In conclusion, K appeared to be involved in sperm motility in dogs and could be a tool to evaluate sperm function. The prostate contributed several elements to canine seminal plasma. Vasectomy changed Ca concentrations and the protein profile of the seminal plasma. Further studies must be performed to clarify the function of these elements on the in vivo fertility of dogs.