Histopathological findings in the early diagnosis of granulosa cell tumour in bitches

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call‐Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call‐Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call‐Exner body.     

Use of two doses of cloprostenol in different intervals for estrus synchronization in hair sheep under tropical conditions

Tropical Animal Health and Production - This study evaluated the effect of two doses of prostaglandin at different intervals on reproductive parameters of crossbred ewes. In Experiment 1, 30 ewes...     

Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples

We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples were T. equi-positive in the nested PCR assay, while the real-time PCR assay also detected the parasite in all 46 of the nested PCR-positive samples but did not detect T. equi in the remaining 19 negative blood samples. This quantitative real-time PCR assay provides a valuable tool for fast laboratory diagnostic assessment of T. equi infection in horses.     

Avian pathogenic Escherichia coli MT78 invades chicken fibroblasts

Avian pathogenic Escherichia coli (APEC) are responsible for extraintestinal diseases, called colibacillosis, in avian species. The most severe manifestation of the disease is colisepticemia that usually starts at the respiratory tract and may result in bird death. However, it is not yet clear how APEC cross the respiratory epithelium and get into the bloodstream. In this work, we studied the interaction between 8 APEC strains (UEL31, UEL17, UEL13, UEL29, MT78, IMT5155, IMT2470, A2363) and a chicken non-phagocytic cell, the fibroblast CEC-32 cell line. We investigated the association profile, the invasion capability, the cytotoxicity effect and the induction of caspase-3/7 activation in an attempt to understand the way the pathogen gains access to the host bloodstream. Association to cells was determined after 1 h of infection, while cell invasion was determined after 4 and 24 h of infection. The cytotoxic effect of bacterial infection was measured by lactate dehydrogenase (LDH) release and the activation of the apoptotic program was verified by caspase-3/7 activation. Also, the presence of genes for adhesins, invasins and other related virulence-associated factors was verified by PCR. All bacterial strains showed similarity in relation to adhesion, LDH release and caspase-3/7 activation. However, one APEC strain, MT78, showed high invasion capability, comparable to the invasive Salmonella typhimurium strain SL1344. Since an APEC strain was capable of invading non-phagocytic cells in vitro, the same may be happening with the epithelial cells of the avian respiratory tract in vivo. CEC-32 monolayers can also provide a useful experimental model to study the molecular mechanisms used by APEC to invade non-phagocytic cells.     

3 TESLA MAGNETIC RESONANCE IMAGING OF THE OCCIPITOATLANTOAXIAL REGION IN THE NORMAL HORSE

The aim of this study was to describe the appearance of the ligamentous structures of the occipitoatlantoaxial (OAA) region in the normal horse by 3 tesla (3T) magnetic resonance imaging (MRI). The MRI images of the longitudinal odontoid ligament, tectorial membrane, dorsal and ventral atlantoaxial ligaments, dorsal atlantooccipital membrane with its reinforcing ligaments, and the lateral atlantooccipital ligaments of 10 horse cadavers were evaluated. All ligaments and membranes were identified in all planes, except for the lateral atlantooccipital ligament in the sagittal plane due to its cranioventrolateral course. All were iso to mildly hypointense to musculature of the neck in T1W with the exception of the tectorial membrane that was moderately hypointense; moderately hypointense in PD‐SPIR, and markedly hypointense (isointense to cortical bone) in T2W. The PD‐SPIR was the best sequence to identify all ligaments and membranes from their cranial and caudal attachments. The longitudinal odontoid ligament, ventral atlantoaxial ligament, and reinforcing bands of the dorsal atlantooccipital membrane presented a characteristic striped heterogeneous signal behavior thought to be due to fibrocartilaginous content. The remaining ligaments and membranes showed homogeneous signal intensity. Special anatomical features in this species such as the fan‐shaped longitudinal odontoid ligament, absence of the transverse ligament and presence of the ventral atlantoaxial ligament were documented. Ligamentous structures that stabilize the equine OAA region were described with MRI in this study and these findings could serve as an anatomic reference for those cases where instability of this region is suspected.     

Caudal epidural anesthesia in mares after bicarbonate addition to a lidocaine–epinephrine combination

Objective

To investigate the nociceptive and clinical effects of buffering a lidocaine–epinephrine solution with sodium bicarbonate in caudal epidural block in mares.

Existence of two O-serotypes in the fish pathogen Pseudomonas anguilliseptica

The serological characteristics of a group of 32 Pseudomonas anguilliseptica strains isolated in Spain from seabream (Sparus aurata) and turbot (Scophthalmus maximus) were compared with a total of 18 collection strains of this bacterial species with different geographical and host origin. The employment of different techniques, including slide agglutination, microagglutination and dot blot, allowed us to establish two serological groups, one comprising practically all the eel isolates, and the other including the majority of isolates from other fish species. The study of the lipopolysaccharides (LPS) and outer membrane proteins (OMP) corroborated these results, indicating that the serological differences among strains are due to the somatic antigen and not to antigenic determinants of protein nature. Therefore, a serological scheme of two "O" serotypes is proposed for P. anguilliseptica. The results obtained will be of importance for epidemiological studies as well as for the design of adequate vaccine formulations.     

The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa

To try new acrosomal probes for assessing ram spermatozoa, we compared the LysoSensor^? probe, which labels acidic organelles, with the frequently used peanut agglutinin acrosomal probe (PNA‐PE; phycoerythrin as fluorescent moiety). The previous microscopic observations showed a lack of relationship of LysoSensor^? with acrosomal status. Semen was obtained from five rams and frozen in four pools. Each pool was analysed carrying out a triple staining propidium ioide/PNA‐PE/LysoSensor^? Green DND‐189 to test acrosome labelling, and a double staining SYBR‐14/PI, to assess sperm viability. Stained samples were analysed by flow cytometry. All measurements were replicated. Data were processed using agreement and repeatability tests. LysoSensor^? labelling did not agree with PNA (mean of differences: 30.8%; coefficient of agreement: 22.6%), confirming microscopic observations. Nevertheless, when LysoSensor^? was compared with SYBR‐14/PI, the agreement was high (mean of differences: ?0.05%; coefficient of agreement: 5.07%). Repeatability of both methods was high and similar. LysoSensor^? did not seem to specifically stain the acrosome, but it may accumulate in the cytoplasm and label viable spermatozoa. Therefore, LysoSensor^? might not be used as an acrosomal probe in ram spermatozoa, but it could be used in other kind of studies, taking advantage of its pH sensitivity.