The objective of this study was to investigate three storage methods and four storage times for rumen sampling in terms of quality and yield of extracted metagenomic DNA as well as the composition of the rumen bacterial community. One Nellore steer fitted with a ruminal silicone-type cannula was used as a donor of ruminal contents. The experiment comprised 11 experimental groups: pellet control (PC), lyophilized control (LC), P-20: pellet stored frozen at -20°C for a period of 3, 6, and 12 months, P-80: pellet stored frozen at -80°C for a period of 3, 6, and 12 months, and L-20: lyophilized sample stored frozen at -20°C for a period of 3, 6, and 12 months. Metagenomic DNA concentrations were measured spectrophotometrically and fluorometrically and ion torrent sequencing was used to assess the bacterial community composition. The L-20 method could not maintain the yield of DNA during storage. In addition, the P-80 group showed a greater yield of metagenomic DNA than the other groups after 6 months of storage. Rumen samples stored as pellets (P-20 and P-80) resulted in lower richness Chao 1, ACE, and Shannon Wiener indices when compared to PC, while LC and PC were only different in richness ACE. The storage method and storage time influenced the proportions of 14 of 17 phyla identified by sequencing. In the P-20 group, the proportion of Cyanobacteria, Elusimicrobia, Fibrobacteres, Lentisphaerae, Proteobacteria, and Spirochaetes phyla identified was lower than 1%. In the P-80 group, there was an increase in the proportion of the Bacteroidetes phylum (p = 0.010); however, the proportion of Actinobacteria, Chloroflexi, SR1, Synergistetes, TM7, and WPS.2 phyla were unchanged compared to the PC group (p > 0.05). The class Clostridium was the most abundant in all stored groups and increased in its proportion, especially in the L-20 group. The rumen sample storage time significantly reduced the yield of metagenomic DNA extracted. Therefore, the storage method can influence the abundance of phyla, classes, and bacterial families studied in rumen samples and affect the richness and diversity index.

The aim of this study was to evaluate and compare the efficacy of B-mode, Doppler, contrast-enhanced ultrasonography (CEUS), and Acoustic Radiation Force Impulse (ARFI) elastography in predicting malignancy in canine mammary masses. This was a prospective cohort study from 2014 to 2016, which included 153 bitches with one or more mammary masses. A total of 300 masses were evaluated by ultrasonography (B-mode, Doppler, CEUS, and ARFI) and subsequently classified as benign or malignant by histopathology. Each ultrasound parameters studied were compared between benign and malignant masses by Chi-square or Student tests and differences were considered significant when P < 0.01. For the variables that proved significant differences were estimated the cut-off point, sensitivity, specificity, accuracy, and area under curve (AUC) by receiver-operating characteristic curve (ROC) analysis in a logistic regression model using histopathological classification as reference, to assess and compare diagnostic performance of each technique. Out of 300 mammary masses evaluated 246 were classified as malignant and 54 as benign. B-mode measurements showed sensitivity 67.9%, and specificity 67.6% as malignancy predictors on canine mammary masses; Doppler indexes systolic (>21.2 m/s) and diastolic velocity (>4.8 m/s) sensitivity 79.2% and specificity 70.8%; CEUS wash-out time ( 2.57 m/s) sensitivity 94.7% and specificity 97.2% In conclusion B-mode and Doppler ultrasound evaluations may assist in malignancy prediction of canine mammary masses with moderate sensitivity and specificity, already the SWV was an great accurate predictor. Therefore, ARFI elastography exam inclusion in veterinary clinic oncology and research is highly recommended, since it allows fast, non-invasive, and complication-free malignancy prediction of canine mammary masses.

The aetiology of septic acute kidney injury (AKI) is not completely elucidated. Early identification of AKI in septic patients is considered to improve survival rate since it allows rapid treatment onset. We evaluated clinical, haematological, urinary, B-mode, spectral Doppler, and contrast-enhanced ultrasound variables in 20 bitches with pyometra as sepsis models and 12 healthy controls. All animals with pyometra presented some degree of renal damage on histological examination; however, sequential organ failure assessment (SOFA) classified only 40% cases with sepsis. AKI derived from systemic infection was identified in 57% of cases with hypoperfusion and in 22% with inflammation, being an affection of multifactorial origin. Among the evaluated parameters, urinary protein/creatinine ratio >0.15, serum albumin <2.94 mg/dL, time-averaged minimum velocity 5.93, pulsatility index >1.53, haematocrit <45%, time-averaged maximum velocity <45.7 cm/s, haemoglobin 12.53 × 10³/uL, and cortical contrast peak intensity 80%. Thus, AKI is a very prevalent condition in septic patients, derived mainly from changes in renal perfusion and inflammation. Additionally, reviewing the SOFA score parameters is suggested to identify renal failure.