Survival of mouse embryos after vitrification depending on the cooling rate of the cryoprotectant solution

The aim of the study was to determine the relationship between the rate of cooling of eight-cell mouse embryos to the temperature of liquid nitrogen (-196 degrees C) and their developmental capacity after thawing on the basis of their ability to leave the zona pellucida ('hatching') during in vitro culturing. Eight-cell embryos were obtained from superovulated female mice and divided into three experimental and one control group. Embryos from the experimental groups were cryopreserved by the vitrification method using ethylene glycol as cryoprotectant. The vitrification protocols used in the study differed in the rate of cooling of the cryoprotectant solution. Embryos from the first group were frozen in conventional 0.25-ml plastic straws, those from the second group in pipetting 'tips', and embryos from the third group, placed in vitrification solution, were introduced dropwise directly into liquid nitrogen. The control group of embryos was cultured in vitro without freezing in a culturing medium in an environment consisting of 95% air and 5% CO2. The developmental capacity of thawed embryos was assessed on the basis of their ability to leave the zona pellucida ('hatching') after three days of in vitro culturing. In the control group 95.1% of embryos 'hatched'. A significantly higher number of embryos that 'hatched' after thawing was observed in the group introduced dropwise directly into liquid nitrogen (60.0%) compared to the group frozen in pipetting 'tips' (37.9%). The group frozen in straws yielded significantly the lowest proportion of 'hatching' embryos (8.1%). These results showed that increasing cooling rates during vitrification of embryos improved their survival.     

Evaluation of jitter by stimulated single-fiber electromyography in normal dogs

Single-fiber electromyography (SFEMG), a technique used to investigate neuromuscular transmission, has been described previously in the pelvic limb of dogs. Because preferential involvement of isolated muscle groups can occur in disorders of neuromuscular transmission, SFEMG was done in the peroneus longus (PL), extensor carpi radialis (ECR), and orbicularis oculi (OO) muscles of 10 adult, clinically normal dogs. Jitter was calculated as the mean absolute value of the consecutive differences in latency of 50 single muscle fiber action potentials after stimulation of intramuscular nerve bundles at the level of the motor point in at least 20 muscle fibers per muscle. Bilateral recordings were performed in 3 dogs. Mean jitter values were determined for each muscle, and differences among muscle groups and among dogs were compared. The upper limits of mean consecutive difference (mean plus 3 standard deviations) for the PL, ECR, and OO muscles were 21.94, 22.53, and 23.39 micros, respectively, and the upper limit of mean consecutive difference for individual muscle fibers in the respective fiber pools was 28.62, 36.39, and 35.68 micros. Jitter values for the ECR and OO were significantly higher than the jitter value for the PL muscle (P < .05). Significant differences among muscles or dogs or between sides were not observed for the ECR. Significant differences among dogs were observed for OO jitter values and were attributed to extremely low jitter values in 1 dog. Significant differences were demonstrated between sides for the PL and were attributed to small sample size. Results of this study provide normative data that can be used in the application of the stimulated SFEMG technique to dogs with suspected disorders of neuromuscular transmission.     

Experimental vascular graft for liver transplantation

Hepatic artery thrombosis is a major cause of graft failure in liver transplantation. Use of donor interponates are common, but results are controversial because of necrosis or thrombosis after rejection. Reperfusion injury, hypoxia and free radical production determinate the survival. The aim of the study was to create an 'ideal' arterial interponate. Autologous, tubular graft lined with mesothelial cells, prepared from the posterior rectus fascia sheath, was used for iliac artery replacement in eight mongrel dogs for six months under immunosuppression. Patency rate was followed by Doppler ultrasound. Eight grafts remained patent and another two are patent after one year. The patency rate was good (median Doppler flow: 370 cm/sec) and there was no necrosis, thrombosis or aneurysmatic formation. The grafts showed viable morphology with neoangiogenesis, appearance of elastin, smooth muscle and endothelial cells. Electron microscopy showed intact mitochondrial structures without signs of hypoxia. Tissue oxygenation was good in all cases with normal (< 30 ng/ml) myeloperoxidase production. In conclusion, this autologous graft presents good long-term patency rate. Viability, arterialisation and low thrombogenicity are prognostic factors indicating usability of the graft in the clinical practice without the risk of rejection. Further investigations such as cell cultures and standardisation are necessary.     

Effects of transcutaneous electrical nerve stimulation on the fatty acid profile of rabbit longissimus dorsi muscle (preliminary report)

This study was designed to investigate whether transcutaneous electrical nerve stimulation (TENS) of the longissimus dorsi muscle (MLD) of rabbits induces specific proportional changes in the muscle fatty acid composition. Ten 4-week-old Pannon White rabbits were exposed to TENS treatment two times a day, with the following settings: 30 Hz, 20 micros impulse length, 10 mA, 2 x 20 min. After a treatment period of 50 days rabbits were slaughtered and the fatty acid composition of the MLD was determined by gas chromatography. The TENS treatment increased the proportions of linoleic (C18:2 n-6), linolenic (C18:3 n-3) and gondoic acids (C20:1 n-9), compared with the control group. The level of palmitic (C16:0), stearic (C18:0), oleic (C18:1 n-9) and eicosapentaenoic (C20:5 n-3) acids significantly decreased. The proportion of total unsaturated fatty acids significantly increased. On the basis of the results obtained, TENS may have similar effects on the muscle fatty acid profile like physical training. Based on the supposal that the composition of membrane structure was also affected, the electrical stimulation of muscles may have further consequences, e.g. on membrane properties.     

Use of protected methionine (Mepron M 85) in cattle

The ruminal stability of Mepron M 85 and the effect of supplementation with Mepron M 85 on free methionine level of blood were studied in rumen-fistulated cows and rumen- and duodenum-fistulated growing bulls. In five rumen-fistulated cows in situ 69.5% and 64.6% of the methionine content of Mepron M 85 was found after ruminal incubation of 16 h and 24 h, respectively. Daily rations of the rumen-fistulated cows were supplemented with 15.0 g DL-methionine and 17.7 g Mepron M 85, which increased the free methionine level of blood from 13.64 mumol/L to 15.35 and 20.46 mumol/L, respectively, three hours after feeding. In the four rumen- and duodenum-fistulated growing bulls, supplementation with 15.0 g DL-methionine and 17.7 g Mepron M 85 increased the total methionine getting into the duodenum during 24 h from 14.99 g to 16.84 and 20.84 g, respectively. The influence of Mepron M 85 on milk production was studied in 35 pairs of Hungarian Fleckvieh x Holstein-Friesian cows. The animals were coupled on the basis of the number of finished lactations, milk production in the previous lactation, and the date of calving. Daily supplementation of 18.0 g Mepron M 85 increased daily milk production significantly (p < 0.05), by 1.24 litres. Milk fat content also increased significantly (from 3.10% to 3.19%, p < 0.05) in the experimental group. The supplementation did not influence milk protein content.     

Assessing Knowledge Production for Agrosilvopastoral Systems in South America

In recent decades agroforestry has undergone significant development in Latin America. South America generates the most scientific research on the topic in the region. We conducted a comprehensive review and analysis of knowledge production for South American agroforestry that includes livestock grazing, known as agrosilvopastoralism (AS), examining how different sociopolitical factors such as sources of funding, institutional priorities, and international cooperation can bias the direction and objectives of scientific research. We assessed the major attributes of scientific publications on the topic (25 articles per country; overall n = 210) and the potential factors underlying the processes of research and development in the region. The tree component was the most studied, while the livestock component received less attention. Studies were mainly focused on the production of goods and services (monetary or nonmonetary approaches), except in Brazil, where conservation was the major study objective. Stakeholders were involved in more than half of the studies (60%), and they were mostly ranchers and local producers. More than half (70%) of the studies offered recommendations based on their results, and such recommendations were mostly concerned with the management of agrosilvopastoral system components. In general, studies were led just as often by local as foreign first authors and coauthored by more than three people as part of interinstitutional working groups. Brazil, Argentina, Colombia, and Chile had more frequent cooperation among institutions and countries but mainly used their own funding. In contrast, Bolivia, Ecuador, and Peru had almost 100% of their studies supported by foreign countries (North America and Europe). Countries with greater internal funding of research generated more long-term studies. Besides this, two clear trends were detected: 1) conservation and social aspects were mainly supported by sources from external countries led by foreign principal investigators, and 2) production issues were supported from sources within countries and supported high levels of cooperation among institutions.     

Role of leptin in farm animals: a review

The discovery of hormone leptin has led to better understanding of the energy balance control. In addition to its effects on food intake and energy expenditure, leptin has now been implicated as a mediator of diverse physiological functions. Recently, leptin has been cloned in several domestic species. The sequence similarity suggests a common function or mechanism of this peptide hormone across species. Leptin receptors are expressed in most of tissues, which is consistent with the multiplicity of leptin functions. The main goal of this review was to summarize knowledge about effect of leptin on physiology of farm animals. Experiments point to a stimulatory action of leptin on growth hormone (GH) secretion, normal growth and development of the brain. Surprisingly, leptin is synthesized at a high rate in placenta and may function as a growth factor for fetus, signalling the nutritional status from the mother to her offspring. Maturation of reproductive system can be stimulated by leptin administration. Morphological and hormonal changes, consistent with a major role of leptin in the reproductive system, have also been described, including the stimulation of the release of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin. Leptin has a substantial effect on food intake and feeding behaviour in animals. Administration of leptin reduces food intake. Its level decrease within hours after initiation of fasting. Leptin also serves as a mediator of the adaptation to fasting, and this role may be the primary function for which was the molecule evolved.     

RESEARCH PAPER: Segmental dorsolumbar epidural analgesia via the caudal approach using multiple port catheters with ketamine or lidocaine or in combination in cattle

ObjectiveTo determine the analgesic and systemic effects of epidural administration of ketamine, lidocaine or a combination of ketamine/lidocaine in standing cattle.Study designProspective, randomized, experimental trial.AnimalsSix healthy male cattle weighing between 335 and 373 kg.MethodsThe animals received 0.5 mg kg^?1 of ketamine (K), 0.2 mg kg^?1 of 2% lidocaine (L) or 0.25 mg kg^?1 ketamine plus 0.1 mg kg^?1 lidocaine (KL). All the drugs were injected into the dorsolumbar epidural space via a caudal approach through a non‐styletted multiple‐port catheter. Each animal received each treatment at random. Evaluations of analgesia, sedation, ataxia, heart rate, arterial pressure, respiratory rate, skin temperature and rectal temperature were obtained at 0 (basal), 5, 10, 15, 30, 45, 60, 75, 90 minutes after epidural injection, and then at 30‐minute intervals until loss of analgesia occurred. Skin temperature was taken at these intervals up to 60 minutes. All the animals received a standard noxious stimulus; a 4‐point scale was used to score the response. A second scale was used to score ataxia and a third for sedation.ResultsThe duration of analgesia in the upper and lower flanks in cattle was 140 ± 15, 50 ± 14 and 80 ± 22 minutes (mean ± SD) after dorsolumbar epidural KL, K or L, respectively. The cardiovascular changes were within acceptable limits in these clinically healthy cattle.ConclusionsDorsolumbar epidural administration of KL to cattle resulted in longer duration of analgesia of the upper and lower flanks in standing conscious cattle, than the administration of K or L alone.Clinical relevanceFurther research is necessary to determine whether this combination using this technique provides sufficient analgesia for flank surgery in standing cattle.     

Evaluation of an antigen capture ELISA for the early diagnosis of Hypoderma lineatum in cattle under field conditions

An antigen capture or sandwich ELISA (sELISA) was evaluated for the diagnosis of Hypoderma lineatum in cattle under field conditions in northwestern Spain. The kinetics of circulating hypodermin C (HyC) and specific antibodies during the course of natural infestation were determined in a group of 10 Frisian calves. In addition, oesophagi and blood samples were taken from 105 cows at a slaughterhouse in order to compare three methods for the diagnosis of H. lineatum: sandwich ELISA for the detection of the antigen HyC (sELISA), indirect ELISA for the detection of antibodies anti-HyC (iELISA) and the detection of first instars (L1) in the oesophagus. In naturally infested cattle, HyC was present in circulation at low levels during the early and late phases of the infestation. However, in the middle phase, coinciding with the presence of L1 in the oesophagus, two peaks of increased HyC concentration were observed. Specific antibodies increased progressively until the first appearance of larvae in warbles on the back. There was no correlation between antigen or antibody levels and the number of grubs in the back. Prevalence of first instars in the oesophagi of slaughtered cows was 21.9% (23/105). The percentage of cattle that were positive for circulating antigen was slightly higher (24.8%), suggesting the recent destruction of migrating larvae in some animals. However, there was no correlation between the number of L1 and HyC levels. With the iELISA, 79% of the animals were positive to Hypoderma, which means that a high percentage of those animals have been exposed to the parasite but they had no apparent current infestation. The sELISA is a good tool to follow larval development within the host; however, the episodic elevation of HyC levels limits the usefulness of this test for the early diagnosis of Hypoderma under field conditions.