A Survey on Vaccination and Disease Occurrence in Municipal and Non-Profit Animal Shelters in Portugal.

Few studies are available describing animal shelters in Portugal. The aim was to characterize prophylactic measures and disease occurrence in shelters with a questionnaire. The response rates of 67 shelters (42 municipal shelters, 25 associations) were compared by the Fisher's exact test. More veterinarians answered for municipal shelters (98%) than for associations (40%; p < 0.001). Over 80% of the respondents indicated using individual medical records and routine prophylaxis. Excessive length of stay for dogs was reported by 54% of associations and 33% of municipal shelters. Management tools should be promoted to improve the situation. Puppy vaccinations were similar and a final vaccination at 16 weeks was indicated by >33% of shelters. Annual revaccination of dogs was reported more frequently by associations (88%) than municipal shelters (55%; p = 0.02). The three most reported diseases were parvovirus and mange in dogs, upper respiratory disease and panleukopenia in cats, and dermatophytosis in both species. Similar response rates for diagnostic options were obtained by both shelter types, except for distemper. Testing for feline retroviruses was indicated by most shelters (>69%), but only a few (<24%) confirmed positive test results. Clinical diagnoses should be complemented by testing. Additional information on disease occurrence should be obtained by objective monitoring.

The heart to single vertebra ratio: A new objective method for radiographic assessment of cardiac silhouette size in dogs.

Vertebral heart size (VHS) is widely determined in clinical practice as an objective method to assess the cardiac silhouette dimensions. However, a key limitation is that it is difficult to determine VHS in dogs with vertebral alterations. This retrospective, method comparison, observer agreement study sought to overcome this limitation by using the heart-to-single vertebra ratio (HSVR), by evaluating the level of agreement between VHS and HSVR, as well as the intra- and inter-observer agreement for HSVR. Three independent observers retrospectively evaluated thoracic radiographs obtained over a set time period. Exclusion criteria were the presence of alterations of the thoracic spine and the inability to clearly outline the cardiac silhouette. The lengths of the vertebral bodies, from the fourth to eighth thoracic vertebra, and VHS were measured on each radiograph. The HSVR was calculated by dividing the sum of the cardiac long and short axes by the length of each vertebral body. Eighty dogs of different breeds were included in the final analysis. Lin's concordance correlation coefficients revealed strong correlations between VHS and HSVR (0.91-0.96), and the Bland-Altman plots showed low bias (0.01-0.2) between the methods. The mean absolute errors indicated low average magnitudes of error (0.11-0.28). The intraclass correlation coefficients showed good to excellent inter-observer (0.87-0.92; P = 0.000) and intra-observer (0.87-0.99; P < .001) agreement. In the authors' opinion, this new method, which is less time consuming and more objective, could offer a valuable alternative to VHS.

Fatal congenital and traumatic cervical spine injuries in a captive newborn plains zebra (Equus quagga).

Background: In this report, we describe the clinical, macro-, and microscopic findings of a newborn zebra victim of a fatal trauma and its possible causes are discussed in detail. Case Description: A plains zebra (Equus quagga) born in a Portuguese zoo was found in hypothermia and unable to get up. The animal was admitted to the hospital 36 hours later and, after the first hour of treatment and no attempts to get up, it began to convulse and died. At necropsy, subdural hematoma and atlantoaxial dislocation were the main findings. Conclusion: The inability to pinpoint the exact moment of the lesions' onset determines the importance of a thorough surveillance of the periparturient period and parturition of captive animals. Furthermore, literature concerning zebras, their reproduction, and neonatal period is scarce, reinforcing the need to report these cases.

Impact of COVID-19 Restrictions in Portugal: A Questionnaire to Municipal and Animal Association Shelters.

The COVID-19 pandemic has an indirect impact on the health and welfare of animals. The aim of this work was to investigate the effect of COVID-19 on Municipal and Association animal shelters. A questionnaire was sent to 97 Municipal shelters and 65 Associations. Questions focused on public funding, management and animal welfare during COVID-19 restrictions. The response rate was 43.3% (42/97) for Municipal shelters and 38.5% (25/65) for Associations. Municipal shelters (67%) received over 80% of public funding, whereas 68% of the Associations received less than 50%. During the COVID-19 restrictions, financial difficulties were observed by 52% of Associations and 5% of Municipal shelters, and a lack of volunteers was observed by 56% of Associations and 17% of Municipal shelters. Operational difficulties were indicated by 43% of Associations and 12% of Municipal shelters, and a lack of instructions was observed by 31% of Municipal shelters and 4% of Associations. No significant differences were obtained on abandonment, adoption, clinical support, exercise and interaction. Decreased feed supply was reported by 40% of Associations and 5% of Municipal shelters. During the COVID-19 restrictions, Municipal shelters were more affected by the lack of instructions, and Associations were more affected by external factors such as a decrease in feed supply, volunteers and funding. Preparedness and contingency plans seem essential to face unpredicted crises.

Clinicopathological and imaging features of pulmonary alveolar microlithiasis in a dog - a case report.

BACKGROUND: The aetiology of pulmonary alveolar microlithiasis (PAM) in animals is still unknown. In humans, this pulmonary disorder is a rare autosomal recessive disorder triggered by a mutation in the gene SLC34A2, which causes deposition and aggregation of calcium and phosphate in the pulmonary parenchyma with formation of microliths. Although histopathological examination is required for a definite diagnosis, in humans, imaging modalities such as computed tomography can demonstrate typical patterns of the disease. This is the first description of the computed tomographic (CT) features of a histologically confirmed PAM in dogs. CASE PRESENTATION: The following report describes a case of a 7-year-old female Boxer dog evaluated for paroxysmal loss of muscle tone and consciousness with excitement. The main differential diagnoses considered were syncope, seizures, and narcolepsy-cataplexy. The results of the complete blood count, serum biochemistry panel, urinalysis, arterial blood pressure, echocardiography, abdominal ultrasound, Holter monitoring, and ECG were all within normal limits. Additional exams included thoracic radiographs, head and thorax CT, bronchoalveolar lavage (BAL), and CT-guided cytology. Thoracic radiographs revealed micronodular calcifications in the lungs, with sandstorm appearance. Computed tomography of the thorax showed the presence of numerous mineralized high-density agglomerates of multiple sizes throughout the pulmonary parenchyma, a reticular pattern with ground glass opacity and intense mineralized fibrosis of the pleural lining. Head CT was unremarkable. BAL and CT-guided cytology were inconclusive, but imaging features strongly suggest the diagnosis of PAM, which was histologically confirmed after necropsy. CONCLUSIONS: This case report contributes to the clinicopathological and imaging characterization of pulmonary alveolar microlithiasis in dogs. In this species, the diagnosis of PAM should be considered when CT features evidence a reticular pattern with ground glass opacity and the presence of an elevated number and size of calcifications.

Mineral Composition of Dry Dog Foods: Impact on Nutrition and Potential Toxicity.

Detailed mineral profile of a selection of commercially available complete dry dog foods was determined using ICP-MS (Se, Cu, Mn, Zn, and nonessential trace elements), flame photometry (Na and K) and atomic and molecular spectrophotometry (Ca, P, Mg, and Fe). The contribution of ingredients to the mineral composition was correlated to the food market segment. Results showed an oversupply of essential elements due to the energy density effect on feed intake. Additives contributed from 40.8 to 55.1% to the total trace elements contents. With the exception of Se, all trace elements were supplied above the nutritional requirements of adult dogs. Legal limits of Cu, Se, and Zn were surpassed. The content of nonessential trace elements included values in the range of nanograms to micrograms per kg, without surpassing safe upper limits. This work brings awareness to the need to find supplementation strategies that ensure nutritional adequacy and avoid waste.

Acute intraparenchymal cerebral haemorrhage in an Iberian golden eagle - a case report.

BACKGROUND: In birds there are reports of intracranial lesions but not of the clinical, computed tomographic and histopathologic features of acute intraparenchymal cerebral haemorrhage in Iberian golden eagle. CASE PRESENTATION: The following report describes a case of a 30-year-old Iberian golden eagle (Aquila chrysaetos homeyeri) with no history of trauma, presented with acute opisthotonus, left head tilt and circling, anisocoria, positional nystagmus, and ataxia. The main differential diagnosis were hypovitaminosis B or E and intracranial disease due to trauma, infection, toxins or masses. A computed tomography (CT) of the head was performed with an 8-slices scanner and evidenced a hyperdense (63-65 HU) non-enhancing homogeneous well delineated round area in the midbrain, with 6 mm in its highest diameter. The attenuation values and the non-enhancing nature of the lesion strongly suggested the diagnosis of acute intraparenchymal haemorrhage, which was histologically confirmed after necropsy. CONCLUSIONS: In birds with a central neurological dysfunction, the diagnosis of acute brain haemorrhage should be considered when the CT evidences a non-enhancing, homogeneous, well circumscribed hyperattenuated round area.

Diferenças na frequência de osteoporosoe entre densitometrias ósseas (DXA) realizadas no Sistema Único de Saúde (SUS) e na Saúde Suplementar (SS) / Differences in frequency of the osteoporosis in bone desitometry performed in national public health system and supplemental heath

Objetivos: Avaliar diferenças no diagnóstico de osteoporose e nos fatores de risco para baixa massa óssea entre densitometrias realizadas pelo Sistema Único de Saúde (SUS) e na Saúde Suplementar (SS). Métodos: Foram analisados retrospectivamente 592 exames de densitometria (anamnese e laudo). Obteve-se dados de sexo, idade, raça, fatores de risco, diagnóstico de osteoporose e especialidade do médico solicitante. Utilizou-se a estatística descritiva e o teste do qui-quadrado. Resultados: Os pacientes do SUS apresentaram maior frequência de osteoporose (54% versus 33%, qui-quadrado=22,63; p<0,0001) e maior número de fatores de risco por indivíduo (qui-quadrado=2.83; p<0,05). A reumatologia foi a especialidade médica que mais requisitou o exame em ambos os grupos. Conclusões: O diagnóstico de osteoporose foi mais frequente nas DXA realizadas no SUS, com maior número de fatores de risco por indivíduo neste grupo.

Objectives: To evaluate differences in diagnosis of osteoporosis and risk factors for low bone mass between exams performed in national public health system (SUS) and supplemental health. Methods: Five hundred ninety two exams (medical records and reports) were analysed. Data on gender, age, race, risk factors, diagnosis of osteoporosis and medical speciality that required the DXA was obtained. Results: Patients who performed DXA in SUS had higher frequency of osteoporosis (54% versus 33%, chi-square = 22.63; p<0,0001) and a higher number of risk factors (chi-square = 2,83; p<0,05). Rheumatology was the medical specialty that most ordered DXA in both groups. Conclusions: The diagnosis of osteoporosis was more frequent in DXA performed in SUS, with higher number of risk factors per individual in this group.

Evaluation of biomarker canine-prostate specific arginine esterase (CPSE) for the diagnosis of benign prostatic hyperplasia.

BACKGROUND: Benign prostatic hyperplasia (BPH) is the most common canine prostatic disorder. Although most or even all intact male dogs may develop BPH by 5-8 years of age, many show no clinical signs. Taking into account the non-specific character of clinical and ultrasonographic findings, a new diagnostic approach has recently been proposed based on the augmentation of blood canine prostate-specific arginine esterase (CPSE) in hyperplasic dogs. The aim of the present study was to verify CPSE levels in negative controls and hyperplasic dogs, considering cytological findings as the reference method and taking into account the fact that controls were middle-aged intact dogs (median of 5.0 years), contrarily to previous studies carried out with very young control dogs. RESULTS: Significant differences of median CPSE levels were found between controls and hyperplasic dogs (29.1 versus 160.7 ng/mL, respectively); and significant positive correlations were found between median CPSE levels and age or prostatic volume (r = 0.549 and 0.448, respectively; p < 0.001). Sensitivity, specificity, positive and negative likelihood ratios put into evidence the good performance of the test. The agreement between methods was found to be very high, notably between CPSE levels and cytological results (Cohen's kappa coefficients above 0.8). CONCLUSIONS: Considering the results all together, measurement of CPSE is confirmed as a useful and accurate method and should be considered as an alternative or complementary tool to conventional methods for the diagnosis of BPH in middle-aged dogs.

ERBB2 in cat mammary neoplasias disclosed a positive correlation between RNA and protein low expression levels: a model for erbB-2 negative human breast cancer.

Human ERBB2 is a proto-oncogene that codes for the erbB-2 epithelial growth factor receptor. In human breast cancer (HBC), erbB-2 protein overexpression has been repeatedly correlated with poor prognosis. In more recent works, underexpression of this gene has been described in HBC. Moreover, it is also recognised that oncogenes that are commonly amplified or deleted encompass point mutations, and some of these are associated with HBC. In cat mammary lesions (CMLs), the overexpression of ERBB2 (27%-59.6%) has also been described, mostly at the protein level and although cat mammary neoplasias are considered to be a natural model of HBC, molecular information is still scarce. In the present work, a cat ERBB2 fragment, comprising exons 10 to 15 (ERBB2_10-15) was achieved for the first time. Allelic variants and genomic haplotype analyses were also performed, and differences between normal and CML populations were observed. Three amino acid changes, corresponding to 3 non-synonymous genomic sequence variants that were only detected in CMLs, were proposed to damage the 3D structure of the protein. We analysed the cat ERBB2 gene at the DNA (copy number determination), mRNA (expression levels assessment) and protein levels (in extra- and intra protein domains) in CML samples and correlated the last two evaluations with clinicopathological features. We found a positive correlation between the expression levels of the ERBB2 RNA and erbB-2 protein, corresponding to the intracellular region. Additionally, we detected a positive correlation between higher mRNA expression and better clinical outcome. Our results suggest that the ERBB2 gene is post-transcriptionally regulated and that proteins with truncations and single point mutations are present in cat mammary neoplastic lesions. We would like to emphasise that the recurrent occurrence of low erbB-2 expression levels in cat mammary tumours, suggests the cat mammary neoplasias as a valuable model for erbB-2 negative HBC.

Sequence variants and haplotype analysis of cat ERBB2 gene: a survey on spontaneous cat mammary neoplastic and non-neoplastic lesions.

The human ERBB2 proto-oncogene is widely considered a key gene involved in human breast cancer onset and progression. Among spontaneous tumors, mammary tumors are the most frequent cause of cancer death in cats and second most frequent in humans. In fact, naturally occurring tumors in domestic animals, more particularly cat mammary tumors, have been proposed as a good model for human breast cancer, but critical genetic and molecular information is still scarce. The aims of this study include the analysis of the cat ERBB2 gene partial sequences (between exon 17 and 20) in order to characterize a normal and a mammary lesion heterogeneous populations. Cat genomic DNA was extracted from normal frozen samples (n = 16) and from frozen and formalin-fixed paraffin-embedded mammary lesion samples (n = 41). We amplified and sequenced two cat ERBB2 DNA fragments comprising exons 17 to 20. It was possible to identify five sequence variants and six haplotypes in the total population. Two sequence variants and two haplotypes show to be specific for cat mammary tumor samples. Bioinformatics analysis predicts that four of the sequence variants can produce alternative transcripts or activate cryptic splicing sites. Also, a possible association was identified between clinicopathological traits and the variant haplotypes. As far as we know, this is the first attempt to examine ERBB2 genetic variations in cat mammary genome and its possible association with the onset and progression of cat mammary tumors. The demonstration of a possible association between primary tumor size (one of the two most important prognostic factors) and the number of masses with the cat ERBB2 variant haplotypes reveal the importance of the analysis of this gene in veterinary medicine.

Sequence variation and mRNA expression of the TWIST1 gene in cats with mammary hyperplasia and neoplasia.

In humans, a germline mutation (c.309C>G) in the TWIST1 oncogene may predispose to breast cancer and its expression has been associated with tumour progression and metastasis. In this study, the feline TWIST1 gene was screened for sequence variations in 37 neoplastic and eight hyperplastic mammary gland lesions from cats. In addition, mRNA levels were examined in 15 mammary tumours and three cases of mammary hyperplasia by quantitative real-time reverse-transcriptase PCR. Feline mammary carcinomas had significantly lower levels of expression of TWIST1 mRNA than benign mammary tumours. No variations were identified in the TWIST1 coding region in feline mammary tumours and the mutation described in humans was not detected. However, two germline variants in the TWIST1 gene intron were identified in four and three carcinomas, respectively: GQ167299:g.535delG and GQ167299:g.460C>T. There was no association between these sequence alterations and TWIST1 mRNA levels.

TWIST1 Gene: First Insights in Felis catus.

TWIST1 is thought to be a novel oncogene. Understanding the molecular mechanisms regulating the TWIST1 gene expression profiles in tumor cells may give new insights regarding prognostic factors and novel therapeutic targets in veterinary oncology. In the present study we partially isolated the TWIST1 gene in Felis catus and performed comparative studies. Several primer combinations were used based on the alignments of homologous DNA sequences. After PCR amplification, three bands were obtained, purified and sequenced. Several bioinformatic tools were utilized to carry out the comparative studies. Higher similarity was found between the isolated TWIST1 gene in Felis catus and Homo sapiens (86%) than between Homo sapiens and Rattus norvegicus or Mus musculus (75%). Partial amino acid sequence showed no change in the four species analyzed. This confirmed that coding sequences presented high similarity (~96%) between man and cat. These results give the first insights regarding the TWIST1 gene in cat but further studies are required in order to establish, or not, its role in tumor formation and progression in veterinary oncology.

Standardised B-scan and A-scan echographic evaluation of spontaneous anterior uveal melanomas in the dog.

In humans, A-scan echographic characteristics of choroidal and ciliary body malignant melanomas permit the prediction of histopathological findings in >95% of cases. The aim of this study was to determine the echographic characteristics, in A-mode and B-mode, of 16 spontaneous anterior uveal melanomas in the dog using a 10 MHz sector transducer. The acoustic hallmark of the tumours was their low to medium reflectivity, internal vascularity and hard consistency. All had an irregular shape, with no acoustic shadowing and the regular lesional spikes. In our experience, the most important sonographic differential diagnoses are ciliary body adenocarcinoma and chronic inflammatory lesions. We believe that standardised A-scan echography can be used in the evaluation and diagnosis of a very high percentage of anterior uveal melanomas.