Effect of mitochondrial uncoupling and glycolysis inhibition on ram sperm functionality

Studies have demonstrated the importance of mitochondria to sperm functionality, as the main source of ATP for cellular homoeostasis and motility. However, the role of mitochondria on sperm metabolism is still controversial. Studies indicate that, for some species, glycolysis may be the main mechanism for sperm energy production. For ram sperm, such pathway is not clear. Thus, we evaluated ram sperm in response to mitochondrial uncoupling and glycolysis inhibition aiming to assess the importance of each pathway for sperm functionality. Statistical analysis was performed by the SAS System for Windows, using the General Linear Model Procedure. Data were tested for residue normality and variance homogeneity. A p < .05 was considered significant. Groups treated with the mitochondrial uncoupler Carbonyl cyanide 3 chlorophenylhydrazone (CCCP) showed a decrease in the percentage of cells with low mitochondrial activity and high mitochondrial membrane potential. We also observed that the highest CCCP concentration promotes a decrease in sperm susceptibility to lipid peroxidation. Regardless the lack of effect of CCCP on total motility, this substance induced significant alterations on sperm kinetics. Besides the interference of CCCP on spermatic movement patterns, it was also possible to observe such an effect in samples treated with the inhibitor of glycolysis (2‐deoxy‐d ‐glucose, DOG). Furthermore, treatment with DOG also led to a dose‐dependent increase in sperm susceptibility to lipid peroxidation. Based on our results, we suggest that the glycolysis appears to be as important as oxidative phosphorylation for ovine sperm kinetics as this mechanism is capable of maintaining full motility when most of the cells have a low mitochondrial membrane potential. Furthermore, we found that changes in the glycolytic pathway trough glycolysis inhibition are likely involved in mitochondrial dysfunction and sperm oxidative unbalance.     

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.     

Brainstem auditory-evoked responses in horses with temporohyoid osteoarthropathy

Background: Facial and vestibulocochlear nerve dysfunction occurs commonly in horses with temporohyoid osteoarthropathy (THO); however, auditory dysfunction has not been thoroughly assessed.
Objective: To determine if auditory abnormalities occur in horses with THO.
Animals: Eleven diseased and 8 control horses.
Methods: This is a prospective study in which brainstem auditory-evoked responses (BAER) were recorded in 11 horses diagnosed with THO through neurologic, endoscopic, radiographic, or computed tomographic examinations. BAER findings were compared with those recorded from 8 adult control horses.
Results: All horses with THO were found to have BAER abnormalities that included complete unilateral BAER loss (82%, n = 9/11), partial unilateral BAER loss (18%, n = 2/11) on the most affected side, and contralateral partial BAER loss (46%, n = 5/11). Nine horses had bilateral THO based on diagnostic imaging findings; of these, 5 (56%) horses also had bilateral BAER abnormalities. The complete absence of BAER in affected horses was most consistent with peripheral sensorineural hearing loss. There was a significant association between complete BAER loss and neurologic and diagnostic abnormalities.
Conclusions and Clinical Importance: Auditory abnormalities such as complete or partial BAER loss are common in horses with THO. The BAER test is an objective diagnostic tool that can aid along with other diagnostic modalities in the assessment, management, and follow-up of horses with THO. Furthermore, BAER studies may help to elucidate the pathophysiology of THO in horses.     

Influence of Heat Stress on the Cortisol and Oxidant‐Antioxidants Balance During Oestrous Phase in Buffalo‐Cows (Bubalus bubalis): Thermo‐protective Role of Antioxidant Treatment

In the present study, the effect of heat stress, which is commonly observed in the animals of Upper Egypt area in summer, as well as the effect of antioxidant treatment as a thermo‐protective was examined. In this study, the animals (n = 120) were divided into winter group (n = 40, bred during winter) and summer group (n = 80, bred during summer) as well as, animals in the summer group were divided into first subgroup animals (n = 40) and injected with Viteselen intramuscularly (15 ml) twice weekly for 10 weeks and second subgroup animals (n = 40) were not treated (as control). Serum levels of progesterone (P4), oestradiol (E2), cortisol, superoxide dismutase (SOD), lipid peroxidase (LPO) and nitric oxide (NO) were measured. The pregnancy rate of all animals was detected rectally. The levels of oestradiol and the activity of the antioxidant SOD were decreased in serum of animals in behavioural oestrus during summer as compared with those in winter. During the same time period the levels of oxidants such as LPO and NO were increased in the serum of animals again in the phase of oestrus. In another group of animals treated by intramuscular injection with 15 ml viteselen (antioxidant) twice weekly for 6 weeks during hot months, the activities of serum SOD showed an increase and the levels of oxidants and cortisol decreased. Moreover, the levels of oestradiol were increased during the oestrous behaviour. The pregnancy rate was decreased in animals under heat stress and the pregnancy rate was enhanced dramatically when these animals received antioxidants during the heat stress. This means that the heat‐stress in Upper Egypt may affect the fertility of animals and pregnancy rate and this effect may be through an increased production of free radicals and decreased production of antioxidants as well as increased levels of cortisol. Treatment of animals or supplementation with antioxidants before the beginning of months of heat‐stress and also during the stress period may correct the infertility due to heat‐stress through the decrease in cortisol secretion and a decrease in the oxidative stress. These results resulted in an increase in pregnancy rate in treated animals.     

PREVALENCE OF ANATOMICAL VARIATION OF THE SIXTH CERVICAL VERTEBRA AND ASSOCIATION WITH VERTEBRAL CANAL STENOSIS AND ARTICULAR PROCESS OSTEOARTHRITIS IN THE HORSE

The sixth cervical vertebra (C6) has unique morphology due to a ventral extension from the transverse process known as the ventral lamina. Little information was found regarding the prevalence and clinical relevance of morphologic variations. Aims of this observational, retrospective study were to characterize C6 morphologic variations in a large sample of horses. Cervical radiographic studies of 100 horses were retrieved. Data recorded were signalment, clinical history, morphology of the C6 ventral lamina, presence of articular process osteoarthritis, and presence of static vertebral canal stenosis. Morphologic variations were found in C6 vertebrae for 24/100 horses, with symmetric absence of the ventral lamina in nine horses and asymmetric absence in 15. Anomalous C6 vertebrae were more common in Warmbloods, with 19/55 Warmbloods in the population being affected (P = 0.006). No association was found with sex. There was no significant difference in the mean of the intravertebral sagittal ratios between horses with normal or anomalous C6 vertebrae; however there was a significantly greater proportion of horses with anomalous C6 vertebrae that had an intravertebral sagittal ratio of less than 0.5 at C6 (P = 0.047). There was no association between the morphology of C6 and articular process osteoarthritis. Anomalous C6 vertebrae in our population were associated with a higher likelihood of cervical pain (P = 0.013). Authors propose that morphologic variations in the C6 ventral laminae could be linked to other developmental abnormalities such as vertebral canal stenosis, might affect regional biomechanics and should therefore be considered clinically relevant in horses. Future, controlled prospective studies are needed to test this theory.     

Effects of seasonal variation,group size and sex on the activity budget and diet composition of the addax antelope

Behaviour and diet composition are critical elements in conservation biology within the scope of reintroduction programs. Here we focused on addax (Addax nasomaculatus), a Critically Endangered antelope species, in the Jbil National Park, Tunisia. In this study, we advanced the hypothesis that season, day period, sex and social structure have an effect on the activity budgets and diet composition of addax. Three groups were selected (large group, adult pair and solitary male). Resting, moving, grazing, vigilance and other behaviours were recorded during two seasons (dry and wet) and in the morning and the afternoon. In addition, faecal samples were collected. The results indicate that season and day period were the main factors affecting the activity budgets. The resting behaviour increased from the wet to the dry season, and from morning to afternoon. The grazing and moving behaviours increased during the wet season and decreased from morning to afternoon. Addax select a mixed diet composed of perennial and herbaceous plant species notably during the wet season. In addition, we observed that addax modify their behaviour according to the season and forage availability. This represents an adaptive strategy to survive in a desert climate with a stochastic low-resource environment and depending on ambient climatic conditions.