Somatic Embryogenesis and Plant Regeneration from Tritordeum

Regeneration of plants by somatic embryogenesis from immature embryos of hexaploid tritordeum (AABBH^chH^ch, amphiploid Hordeum chilense×Triticum turgidum conv. durum) and durum wheat (Triticum tergidum) was induced on MS medium supplemented with different 2.4-D concentrations. Well-defined embryoids were formed with a high frequency on the scutellar callus from 1 or 2 weeks onwards and plantlets were developed from them. In the best cases from one single explant more than 100 plants could be obtained. Plants were also regenerated by somatic embryogenesis from inflorescences of Hordeum chilense×Triticum turgiditm conv. durum hybrid and its respective hexa-amphiploid. With regard to callus induction and regenerative ability, evident differences between hexa- and octoploid (H. chilense×T. aestivum) tritordeum were found, the latter showing a very low response.     

Diagnosing feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) infection: an update for clinicians

With the commercial release in Australia in 2004 of a vaccine against feline immunodeficiency virus (FIV; Fel‐O‐Vax FIV®), the landscape for FIV diagnostics shifted substantially. Point‐of‐care (PoC) antibody detection kits, which had been the mainstay for diagnosing FIV infection since the early 1990s, were no longer considered accurate to use in FIV‐vaccinated cats, because of the production of vaccine‐induced antibodies that were considered indistinguishable from those produced in natural FIV infections. Consequently, attention shifted to alternative diagnostic methods such as nucleic acid detection. However, over the past 5 years we have published a series of studies emphasising that FIV PoC test kits vary in their methodology, resulting in differing accuracy in FIV‐vaccinated cats. Importantly, we demonstrated that two commercially available FIV antibody test kits (Witness? and Anigen Rapid?) were able to accurately distinguish between FIV‐vaccinated and FIV‐infected cats, concluding that testing with either kit offers an alternative to PCR testing. This review summarises pertinent findings from our work published in a variety of peer‐reviewed research journals to inform veterinarians (particularly veterinarians in Australia, New Zealand and Japan, where the FIV vaccine is currently commercially available) about how the approach to the diagnosis of FIV infection has shifted. Included in this review is our work investigating the performance of three commercially available FIV PoC test kits in FIV‐vaccinated cats and our recommendations for the diagnosis of FIV infection; the effect of primary FIV vaccination (three FIV vaccines, 4 weeks apart) on PoC test kit performance; our recommendations regarding annual testing of FIV‐vaccinated cats to detect ‘vaccine breakthroughs’; and the potential off‐label use of saliva for the diagnosis of FIV infection using some FIV PoC test kits. We also investigated the accuracy of the same three brands of test kits for feline leukaemia virus (FeLV) diagnosis, using both blood and saliva as diagnostic specimens. Based on these results, we discuss our recommendations for confirmatory testing when veterinarians are presented with a positive FeLV PoC test kit result. Finally, we conclude with our results from the largest and most recent FIV and FeLV seroprevalence study conducted in Australia to date.     

Interrelated effects of temperature, light and oxygen on Amaranthus caudatus L. seed germination

The aim of this study was to investigate the effects of temperature, light and oxygen on the germination of Amaranthus caudatus (var. viridis) seeds. At the highest temperature studied (35°C), seed germination was found to be most rapid and highest in both the dark and the light. Germination was inhibited by continuous white light. The lower the temperature, the greater the inhibitory effect of light. The far-red and blue spectral regions were the most effective in light inhibition. The light effect was also dependent on the percentage of oxygen in the atmosphere around the seeds; it was less marked in O₂-enriched atmospheres. Light/ oxygen dependency resulted from the action of the seed coat and endosperm. Scarified seeds were much less sensitive to light and hypoxia than non-scarified seeds. Effets combinés de la température, de la lumière et de I'oxygène sur la germination des graines d'Amaranthus caudatus L. Le but de ce travail était d'étudier les effets de la température, de la lumiere et de I'oxygène sur la germination des graines d'Amaranthus caudatus (var. viridis). A la temperature la plus élevée utilisée (35°C), la germination était la plus rapide et la plus complète tant à la lumière qu'à 1'obscuritè. La germination était inhibée par la lumière blanche continue. Plus la temperature était basse, plus 1'effet inhibiteur de la lumière était important. Les radiations rouge sombre et bleues du spectre étaient les plus in-hibitrices. L'effet de la lumière dépendait aussi de la teneur de 1'atmosphère en oxygène; il était moins marqué dans des atmosphères enrichies en oxygène. L'interaction lumière/oxygène provenait du tégument et de 1'albumen. Les graines scarifiées étaient beaucoup moins sensibles à la lumière et a 1'hypoxie que les graines non scarifiées. Wechselwirkungen von Temperatur, Licht und Sauerstoff auf die Keimung der Samen von Amaranthus caudatus L. Sowohl im Dunkeln als auch im Licht keimten die Samen von Amaranthus caudatus (var. viridis) bei 35°C, der höchsten Temperatur der Versuche, am schnellsten und meisten. Durch weiβes Dauerlicht wurde die Keimung gehemmt. Je niedriger die Temperatur, desto hölier die Hemmung durch Licht. Im entfernt roten und blauen Spektralbereich war die Lichthemmung am stärksten. Die Wirkung des Lichts hing auch vom Sauerstoffgehalt der die Samen umgebenden Luft ab: Bei O₂₅-Anreiche-rung war sie weniger deutlich. Die Licht/ Sauerstoff-Wechselwirkung wurde von der Samenschale und dem Endosperm beeinfluβt: Geritzte Samen waren gegenüber Licht und O₂-Mangel viel weniger empfindlich als ganze.     

Dormancy of Bromus rubens L. seeds in relation to temperature, light and oxygen effects

Freshly harvested mature Bromus rubens L. seeds are dormant. This dormancy corresponds to the inability of seeds to germinate in the dark at temperatures above 15°C, but is practically not apparent at temperatures between 5 and 15°C. Continuous white light markedly inhibits germination of dormant seeds, even at low irradiances, and reinforces their sensitivity to oxygen deprivation. However, prolonged illumination with white light does not alter subsequent germination in the dark. Breaking of dormancy during dry storage at 20°C results in a widening of the temperature range within which a high rate of germination occurs, and in almost complete disappearance of sensitivity of seeds to light. The structures surrounding the embryo (glumellae, caryopsis envelopes and possibly endosperm) are mainly responsible for dormancy and photosensitivity of whole seeds, probably because they limit the oxygen supply to the embryo. However, the embryo itself appears to play a role in dormancy. The results obtained are discussed in relation to dormancy of cereal seeds, and in relation to field establishment of B. rubens as a weed. La dormance des semences de Bromus rubens L. en relation avec les effets de la température, de la lumière et de l'oxygène Les semences mûres de Bromus rubens L. fraîchement récoltées sont dormantes. Leur dormance correspond à une inaptitude à germer à l'obscuritéà des températures supérieures à 15°C, mais elle ne se manifeste pratiquement pas entre 5 et 15°C. La lumière blanche continue inhibe nettement la germination des semences dormantes, même à de faibles énergies, et renforce leur sensibilitéà la privation d'oxygène. Cependant, un éclairement prolongé n'a aucune conséquence sur la germination ultérieure à l'obscurité. L'élimination de la dormance pendant la conservation au sec, à 20°C, se traduit par un élargissement de la gamme thermique qui assure une bonne germination et par la disparition presque complète de la sensibilité des semences à la lumière. Les structures qui entourent l'embryon (glumelles, enveloppes du caryopse et peut-être albumen) sont, en grande partie, responsables de la dormance et de la photosensibilité des semences entières, sans doute parce qu'elles limitent l'apport d'oxygène à l'embryon. Toutefois, l'embryon lui-même semble participer à la dormance. Les résultats obtenus sont discutés par comparaison avec la dormance des semences des céréales et en relation avec l'implantation, dans les cultures, de B. rubens en tant que mauvaise herbe. Dormanz der Samen von Bromus rubens L. in Bezug auf Temperatur, Licht und Sauerstoffversorgung Frisch geerntete Samen von Bromus rubens L. sind im Dunkeln bei Temperaturen über 15°C dormant, während diese Dormanz bei Temperaturen zwischen 5 und 15°C praktisch nicht vorliegt. Ständiges weißes Licht hemmt die Keimung dormanter Samen deutlich, sogar bei niedriger Intensität, und verstärkt ihre Empfindlichkeit gegenüber Sauerstoffmangel. Ständige Beleuchtung mit weißem Licht verändert jedoch nicht die folgende Dunkelkeimung. Das Brechen der Dormanz während der trockenen Lagerung bei 20 °C führt zu einem vergrößerten Temperaturbereich, in dem die Samen gut keimen, und läßt ihre Lichtempfindlichkeit fast vollständig verschwinden. Die Strukturen um den Embryo (Spelzen, Schale der Caryopse und möglicherweise Endosperm) sind hauptsächlich für die Dormanz und die Lichtempfindlichkeit des ganzen Samens verantwortlich, vermutlich deshalb, weil sie die Sauerstoffversorgung des Embryos einschränken. Aber auch der Embryo selbst scheint bei der Dormanz eine Rolle zu spielen. Die Untersuchungsergebnisse werden im Vergleich zur Dormanz von Getreidesamen und hinsichtlich des Vorkommens von Bromus rubens als Unkraut diskutiert.     

Arterial Vascularization of the Gastrointestinal Tract of the Pampas Deer (Ozotoceros bezoarticus,Linnaeus, 1758)

Based on gross dissection of fifteen adult animals (11 females, 4 males), we described the arterial supply of the stomach and intestines of the pampas deer (Ozotoceros bezoarticus), a South American endangered species. The coeliac artery emitted the splenic, left gastric and hepatic arteries. The splenic artery directed towards the spleen, and the right ruminal artery, which is its only collateral directed towards the stomach, being the main artery of the rumen. The left gastric artery gave origin to the left ruminal, the reticular and the left gastroepiploic arteries. The left gastroepiploic artery originated the reticular accessory artery. Both arteries, gastric and left gastroepiploic, anastomosed their right counterparts derived from the hepatic artery on the curvatures of the abomasum. The cranial mesenteric artery irrigated the second half of the duodenum until the beginning of the descending colon. The thickest branch emitted by the cranial mesenteric artery was the ileocolic artery, which was destined to the ascending colon, caecum and ileum. The colic branches and the right colic arteries were irradiated on the right surface of the spiral loop of the ascending colon and distributed to both centripetal and centrifugal coils of the ascending colon; the colic branches were also anastomosed with the last jejunals and ileals and with the right colic arteries. There were no variations in the origin of any of the main branches derived from the coeliac and cranial mesenteric arteries. This species had a basic pattern of arterial distribution similar to small domestic ruminants.     

Molecular Epidemiology of Mycobacterium bovis in Humans and Cattle

Bovine tuberculosis (bTB), caused by Mycobacterium bovis (M. bovis), is a serious re‐emerging disease in both animals and humans. The evolution of the Multi‐ and Extensively drug‐resistant M. bovis strains (MDR‐TB and XDR‐TB) represents a global threat to public health. Worldwide, the disease is responsible for great economic losses in the veterinary field, serious threat to the ecosystem, and about 3.1% of human TB cases, up to 16% in Tanzania. Only thorough investigation to understand the pathogen's epidemiology can help in controlling the disease and minimizing its threat. For this purpose, various tools have been developed for use in advanced molecular epidemiological studies of bTB, either alone or in combination with standard conventional epidemiological approaches. These techniques enable the analysis of the intra‐ and inter‐species transmission dynamics of bTB. The delivered data can reveal detailed insights into the source of infection, correlations among human and bovine isolates, strain diversity and evolution, spread, geographical localization, host preference, tracing of certain virulence factors such as antibiotic resistance genes, and finally the risk factors for the maintenance and spread of M. bovis. They also allow for the determination of epidemic and endemic strains. This, in turn, has a significant diagnostic impact and helps in vaccine development for bTB eradication programs. The present review discusses many topics including the aetiology, epidemiology and importance of M. bovis, the prevalence of bTB in humans and animals in various countries, the molecular epidemiology of M. bovis, and finally applied molecular epidemiological techniques.     

Achievements and future perspectives of embryo transfer technology in pigs

Commercial embryo transfer (ET) has unprecedented productive and economic implications for the pig sector. However, pig ET has been considered utopian for decades mainly because of the requirements of surgical techniques for embryo collection and embryo deposition into recipients, alongside challenges to preserve embryos. This situation has drastically changed in the last decade since the current technology allows non‐surgical ET and short‐ and long‐term embryo preservation. Here, we provide a brief review of the improvements in porcine ET achieved by our laboratory in the past 20 years. This review includes several aspects of non‐surgical ET technology and different issues affecting ET programmes and embryo preservation systems. The future perspectives of ET technology are also considered. We will refer only to embryos produced in vivo since they are the only type of embryos with possible short‐term use in pig production.     

The Effect of Glycerol Concentrations on the Post‐thaw In Vitro Characteristics of Cryopreserved Sex‐sorted Boar Spermatozoa

The objective of this study was to optimize protocols for the cryopreservation of sex‐sorted boar spermatozoa. In the experiment 1, we evaluated the effects of a standard boar sperm cryopreservation procedure (3% final glycerol concentration) on the in vitro characteristics of sex‐sorted sperm frozen at low sperm concentrations (20 × 10⁶ sperm/ml; S20 group). Non‐sorted spermatozoa frozen at 1000 × 10⁶ (C1000 group) and 20 × 10⁶ (C20 group) sperm/ml were used as the freezing control groups. In experiment 2, the effects of different final glycerol concentrations (0.16%, 0.5%, 1.0%, 2.0% and 3.0%) on post‐thaw quality of the S20 and C20 groups were evaluated. In both experiments, the samples were evaluated prior to freezing (5°C) and at 30, 90 and 150 min after thawing. Experiment 1 indicated that freezing sperm at low concentrations decreased (p < 0.05) the total motility (TM) and progressive motility (PM) at 90 and 150 min after thawing regardless of whether the sperm were sorted or not. However, the sperm membrane integrity was not affected at any evaluation step. Inexperiment 2, significant effects on the TM and PM because of increased glycerol concentrations in the S20 and C20 groups were observed only at 90 and 150 min after thawing. The samples frozen in 3% glycerol showed lower (p < 0.05) TM and PM values when compared to those frozen in the presence of 0.5% and 1% glycerol. In both experiments, non‐sorted control samples displayed higher percentages of spermatozoa with damaged DNA than sorted spermatozoa. In conclusion, the optimization of cryopreservation conditions by decreasing the glycerol concentrations can improve post‐thaw motility of sex‐sorted spermatozoa frozen at low concentrations.