Clinical signs associated with prostatic disorders in canines: Retrospective study in Uruguay (2011-2019).

Background: Prostate disease represents about 0.7% of diseases in canines. The main diagnosed pathology is benign prostatic hyperplasia (BPH). However, the reports that study the association of a certain clinical sign with a specific prostate disease are scarce. Aim: The main objective of this study was to evaluate the clinical relevance of the most commonly observed clinical signs associated with the different prostatic disorders in canines admitted to the hospital of the Facultad de Veterinaria-Universidad de la República between 2011 and 2019. Methods: This retrospective study included 7,729 male canines treated at the hospital de la Facultad de Veterinaria-Universidad de la República (Montevideo, Uruguay) between 2011 and 2019. 289 canines with a presumptive/definitive diagnosis of prostate diseases were selected, recording the presence/absence of associated clinical signs. Results were reported in terms of odds ratios (ORs) using logistic regression (p < 0.05). Results: The five most frequently reported clinical signs were tenesmus (34%), anorexia (32%), lethargy (27%), prostatomegaly or pain during rectal examination (25%), and abdominal pain from palpation (22%). Diarrhea (3.39 vs. 0.33 OR), anorexia (2.07 vs. 0.39 OR), weight loss (2.27 vs. 0.27 OR), hematuria (3.25 vs. 0.44 OR), and urinary incontinence (2.96 vs. 0.33 OR) indicated a highest predictive value (p < 0.05) with prostatitis versus BPH, respectively. Being weight loss, the clinical sign is more frequently associated with neoplasia (20.2 OR, p = 0.002). Conclusion: This study shows that there are clinical signs with a higher degree of association for certain canine prostatic disorders than others.

Detection and genome characterisation of SARS-CoV-2 P.6 lineage in dogs and cats living with Uruguayan COVID-19 patients.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in domestic animals have occurred from the beginning of the pandemic to the present time. Therefore, from the perspective of One Health, investigating this topic is of global scientific and public interest. OBJECTIVES: The present study aimed to determine the presence of SARS-CoV-2 in domestic animals whose owners had coronavirus disease 2019 (COVID-19). METHODS: Nasopharyngeal and faecal samples were collected in Uruguay. Using quantitative polymerase chain reaction (qPCR), we analysed the presence of the SARS-CoV-2 genome. Complete genomes were obtained using ARTIC enrichment and Illumina sequencing. Sera samples were used for virus neutralisation assays. FINDINGS: SARS-CoV-2 was detected in an asymptomatic dog and a cat. Viral genomes were identical and belonged to the P.6 Uruguayan SARS-CoV-2 lineage. Only antiserum from the infected cat contained neutralising antibodies against the ancestral SARS-CoV-2 strain and showed cross-reactivity against the Delta but not against the B.A.1 Omicron variant. MAIN CONCLUSIONS: Domestic animals and the human SARS-CoV-2 P.6 variant comparison evidence a close relationship and gene flow between them. Different SARS-CoV-2 lineages infect dogs and cats, and no specific variants are adapted to domestic animals. This first record of SARS-CoV-2 in domestic animals from Uruguay supports regular surveillance of animals close to human hosts.

Detection of high antibodies titers against rat leukemia virus in an outbreak of reproductive disorders and lymphomas in Wistar rats.

Young female Wistar rats from a specific pathogen free breeding colony presented an outbreak of infertility along with neurological symptoms and malignant lymphomas. We evaluated the presence and the potential role of the rat leukemia virus (RaLV) in the disease because these clinical signs could be compatible with a retrovirus. RaLV is a mammalian type C endogenous retrovirus initially isolated from in vitro Sprague-Dawley rat embryo cultures. There are no reports of clinical disease in rats associated with this virus, and little is known about its interaction with the host. Using reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, we studied the synthesis of the viral particles and the development of an immune response against the virus in this rat colony. The results showed that healthy and diseased Wistar rats synthetized viral RNA but only diseased animals developed a detectable immune response against RaLV envelop protein. Furthermore, rats with lymphomas tended to have higher titers of antibodies against RaLV epitopes than those with infertility or neurological symptoms. The results suggest that increases in the RaLV infectious particle loads could be involved in the development of lymphomas in young rats. The potential causes of RaLV reactivation are discussed.

Post-castration prostatic involution: A morphometric and endocrine study of healthy canines and those with benign prostatic hyperplasia.

The objective of this study was to evaluate the effect of castration on dimensions of the prostate by B-mode ultrasound for 90 days, as well as to evaluate changes in serum concentrations of testosterone and oestradiol in clinically healthy canines and those with benign prostatic hyperplasia (BPH). Three groups of animals were used: healthy canines castrated on day 0 (HCC, n = 8), canines with a diagnosis of benign prostatic hyperplasia (BPHC, n = 9) castrated on day 0 and healthy uncastrated canines (IHCC, n = 10). Prostate size was obtained by B-mode ultrasound, while blood samples were collected (days 0, 7, 14, 28, 60 and 90 post-castration) to determine the serum testosterone and oestradiol concentration. Castration did not affect serum oestradiol concentrations. Castration led to a decrease in serum testosterone (reaching values <20 ng/dl in the first week, p < .05), causing a gradual decrease in dimensions of prostate parameters relative to body weight (p < .05). Canines with BPHC had similar prostate dimensions as the IHCC group 2 weeks post-castration. In relation to the prostate's relative parameters, the decrease with time was greater in canines with BPHC than those in the HCC group. When evaluating parameters as a percentage of relative change, the decrease with time was similar in both castrated groups, except for relative length percentage, which presented a more pronounced decline in BPHC canines than HCC canines. In conclusion, castration is an effective, fast and long-lasting treatment for canine patients with BPH, as it causes a rapid decrease in blood testosterone concentrations and a decrease in glandular volume, but does not affect serum oestradiol concentrations. This study did not determine the mechanisms of action, but allowed us to observe (based on morphometric variables) that characteristics of prostate involution after castration differed between canines with BPH and healthy canines.

Detection and genome characterisation of SARS-CoV-2 P.6 lineage in dogs and cats living with Uruguayan COVID-19 patients

BACKGROUND Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in domestic animals have occurred from the beginning of the pandemic to the present time. Therefore, from the perspective of One Health, investigating this topic is of global scientific and public interest. OBJECTIVES The present study aimed to determine the presence of SARS-CoV-2 in domestic animals whose owners had coronavirus disease 2019 (COVID-19). METHODS Nasopharyngeal and faecal samples were collected in Uruguay. Using quantitative polymerase chain reaction (qPCR), we analysed the presence of the SARS-CoV-2 genome. Complete genomes were obtained using ARTIC enrichment and Illumina sequencing. Sera samples were used for virus neutralisation assays. FINDINGS SARS-CoV-2 was detected in an asymptomatic dog and a cat. Viral genomes were identical and belonged to the P.6 Uruguayan SARS-CoV-2 lineage. Only antiserum from the infected cat contained neutralising antibodies against the ancestral SARS-CoV-2 strain and showed cross-reactivity against the Delta but not against the B.A.1 Omicron variant. MAIN CONCLUSIONS Domestic animals and the human SARS-CoV-2 P.6 variant comparison evidence a close relationship and gene flow between them. Different SARS-CoV-2 lineages infect dogs and cats, and no specific variants are adapted to domestic animals. This first record of SARS-CoV-2 in domestic animals from Uruguay supports regular surveillance of animals close to human hosts.

Hematological and biochemical profiles of canine CD45- T lymphomas are different from other immunophenotypes.

BACKGROUND: Canine multicentric lymphomas are lymphoproliferative malignancies that have increased in recent decades. The patient's treatment and prognosis are determined by the grade, histological type, and lymphoma immunophenotyping. AIM: To investigate the paraclinical signs and survival time in canines with different lymphoma immunophenotypes. METHODS: Over 2 and a half years, 47 untreated dogs were diagnosed with multicentric lymphoma at the Veterinary School Hospital of Uruguay. The disease was clinically and cytologically diagnosed, and immunophenotyping was determined by flow cytometry. After the immunophenotyping, most of the patients were grouped into the following: B (LB), T aggressive (LTCD45+), or T-zone lymphoma (LTCD45-). The patients' haematological values, calcemia, lactate dehydrogenase (LDH) levels, and plasmatic electrophoretic profiles were all determined immediately after that. RESULTS: Of all canine lymphomas, 55.3% were B, 31.9% were LTCD45+, and 10.6% were TCD45-. Only 2.2% were classified as nonB/nonT, and survival time differed between groups. Patients with LTCD45- lymphomas had a mean life span of 641 days after diagnosis, followed by LB (166 days) and LTCD45+ (62 days). Red blood cell count, hematocrit, and hemoglobin levels did not differ between groups. However, the LTCD45- group had significantly higher lymphocyte levels than the LTCD45+ and LB groups (p = 0.01 and 0.006, respectively). Levels of albumin, alpha-1, and alpha-2 globulins did not differ between groups. On the other hand, gamma globulins levels in the LTCD45- were higher than in the other lymphoma groups. The presence of hypercalcemia and high plasma LDH levels were associated with patient severity. Only the TCD45+ group had hypercalcemia although both the LB and TCD45+ groups had elevations in LDH activity. Interestingly, there was a direct relationship between high LDH values (greater than 500 IU/l) and lower survival in TCD45+ lymphomas. CONCLUSION: Survival time and hematological and biochemical patterns differed among canine lymphomas immunophenotypes. Patients of LTCD45- phenotype showed higher lymphocyte counts and gamma globulin levels and more prolonged survival. Serum LDH activity may provide additional prognostic information in high-grade T-cell lymphoma.

Serial collection method of dog saliva: Effects of different chemical stimulants on behaviour, volume and saliva composition.

The objective of this study was to evaluate different chemical stimulants with different flavours such as acids (citric and acetic), sweet (sucrose) and salty (sodium chloride) applied to cotton rolls and compare their effects on the volume, pH and protein concentrations of the saliva collected and the behaviour of dogs during sampling management. As an additional objective, serum cortisol concentrations of saliva samples obtained with or without citric acid and with or without previous pH adjustment were compared. Five clinically healthy were randomly assigned to one of 5 treatments with cottons with different substances: 1) control, 2) citric acid, 3) acetic acid, 4) sodium chloride, 5) sucrose. Each dog received one treatment per day, and in 5 days, all dogs were tested with the five treatments. On each day, cottons were applied to dogs at times 0, 20, 40, 60 and 80 minutes. The cottons with citric acid generated more volume than the rest of the treatments (p<0.0001), and sodium chloride generated more volume than the control and acetic acid (p≤0.03). Cottons with citric acid generated lower pH of saliva than the rest of the treatments (p<0.0001). Cottons with acetic acid generated lower pH than control, sodium chloride and sucrose (p<0.0001). There were no differences in cortisol concentrations between the control samples and those obtained with citric acid, nor between these same samples with and without pH adjusted with buffer. The concentration of proteins in saliva and excitement degree did not change with treatment. Citric acid was more palatable than the rest of the treatments (p<0.0001). Sodium chloride and sucrose were more palatable than control (p<0.05). In conclusion, citric acid was the chemical stimulant that generated greater volume of saliva and greater palatability in dogs. Although the pH of the saliva obtained with citric acid was clearly acidic, its acidic pH did not affect the determination of cortisol by chemiluminescence or RIA. Sodium chloride and sucrose allowed to obtain high volumes of saliva and were more palatable than the control, which can be other interesting options to obtain saliva in case of not being able to use citric acid.

Extra-gonadal steroids modulate non-breeding territorial aggression in weakly electric fish.

The neuroendocrine control of intraspecific aggression is a matter of current debate. Although aggression in a reproductive context has been associated with high levels of circulating androgens in a broad range of species, it has also been shown to occur during the non-breeding season when gonads are regressed and plasma steroid hormone levels are low. In mammals and birds the aromatization of androgens into estrogens plays a key role in the regulation of aggression in both the breeding and non-breeding seasons. This is the first study in a teleost fish to explore the role of steroids in the modulation of non-breeding aggression. Gymnotus omarorum is a highly aggressive teleost fish that exhibits aggression all year-round. We analyzed male-male non-breeding agonistic behavior, compared circulating 11-Ketotestosterone (11-KT) levels between dominants and isolated males, assessed the regulatory role of aromatization of androgens into estrogens, and evaluated the gonads as a source of these sex steroids. We found that high levels of aggression occurred in the non-breeding season despite low plasma 11-KT levels, and that there was no difference in 11-KT levels between dominant and isolated males. We demonstrated that acute aromatase inhibition decreased aggression, distorted contest dynamics, and affected expected outcome. We also found that castrated individuals displayed aggressive behavior indistinguishable from non-castrated males. Our results show, for the first time in teleost fish, that territorial aggression of G. omarorum during the non-breeding season depends on a non-gonadal estrogenic pathway.

Immunohistochemical determination of estrogen receptor-α in canine vaginal biopsies throughout proestrus, estrus, and early diestrus.

The aim of this study was to describe the presence of estrogen receptor-α (ERα) in several vaginal histological compartments in healthy adult bitches throughout three estrous cycle stages (proestrus, estrus, and early diestrus) and to relate ERα presence with serum progesterone and estradiol-17ß concentrations. For this purpose, serial blood samples and vaginal biopsies were taken from five bitches every 48 hours, starting at the clinical onset of proestrus, marked by the beginning of serosanguineous vaginal secretion. Serum progesterone and estradiol-17ß concentrations were determined by RIA, whereas detection of steroid receptors was carried out through immunohistochemistry. Subjective image analysis was conducted by two independent observers in the following histological compartments: superficial, intermediate, and deep epithelia and superficial (loose) and deep (dense) stroma (connective tissue). Nuclear ERα immunoreactivity was detected in every histological compartment and estrous cycle stage studied. ERα expression varied among histological compartments and during stages of the cycle. Receptor expression was associated with estradiol-17ß and progesterone serum profiles. Most relevant cyclic changes were detected in the superficial and deep epithelia and in the dense connective tissue. The highest ERα expression was detected during diestrus, although each compartment had a different pattern throughout the other cycle stages. Thus, vaginal ERα expression in the bitch varied throughout proestrus, estrus, and early diestrus according to the histological compartment involved.

Cortisol secretion after adrenocorticotrophin (ACTH) and dexamethasone tests in healthy female and male dogs.

BACKGROUND: For the conclusive diagnosis of Cushing's Syndrome, a stimulating ACTH test or a low suppressive Dexamethasone test is used. Reports in other species than the dog indicate that plasma cortisol concentration after ACTH administration is affected by gender. We investigated the effect of gender on the cortisol response to ACTH and Dexamethasone tests in dogs. METHODS: Seven healthy adult Cocker Spaniels (4 females and 3 males) were assigned to a two by two factorial design: 4 dogs (2 females and 2 males) received IV Dexamethasone 0.01 mg/kg, while the other 3 dogs received an IV saline solution (control group). Two weeks later the treatments were reversed. After one month, ACTH was given IV (250 microg/animal) to 4 dogs (2 female and 2 males) while the rest was treated with saline solution (control group). Cortisol concentrations were determined by a direct solid-phase radioimmunoassay and cholesterol and triglycerides by commercial kits. RESULTS AND DISCUSSION: No effect of treatment was observed in metabolite concentrations, but females presented higher cholesterol concentrations. ACTH-treated dogs showed an increase in cortisol levels in the first hour after sampling until 3 hours post injection. Cortisol concentrations in Dexamethasone-treated dogs decreased one hour post injection and remained low for 3 hours, thereafter cortisol concentrations increased. The increase in cortisol levels from one to two hours post ACTH injection was significantly higher in females than males. In Dexamethasone-treated males cortisol levels decreased one hour post injection up to 3 hours; in females the decrease was more pronounced and prolonged, up to 5 hours post injection. CONCLUSION: We have demonstrated that cortisol response to ACTH and Dexamethasone treatment in dogs differs according to sex.