Effect of Different Manganese Concentrations during in vitro Maturation of Bovine Oocytes on DNA Integrity of Cumulus Cells and Subsequent Embryo Development

Manganese (Mn) is a trace element present in forages and cereals, and its concentration depends on soil status. Manganese deficiency in cattle, goats and ewes not only impairs oestrous cycle but reduces calf birth weight. The achievement of the first oestrus is delayed, and more attempts are necessary to obtain a successful conception. This study was conducted to investigate the effect of the availability of supplemental Mn during IVM on DNA damage of cumulus cells and total glutathione (GSH) content in oocytes and cumulus cells. The effect of supplementary Mn during IVM on subsequent embryo development was also studied. The results reported here indicate (i) DNA damage in cumulus cells decreased with 0, 2, 5 and 6 ng/ml Mn supplementation during IVM (p < 0.05). (ii) Intracellular GSH‐GSSG content increased (p < 0.01) with different Mn concentrations in oocytes and cumulus cells. Also, cumulus cell number per cumulus oocyte‐complexes (COC) did not differ either before or after IVM. (iii) Addition of Mn to maturation medium resulted in similar cleavage rates (p > 0.05) at 0, 2, 5 and 6 ng/ml Mn. However, subsequent embryo development to blastocyst stage was significantly higher (p < 0.01) in oocytes matured with 5 and 6 ng/ml Mn. (iv) There was also an increase (p < 0.05) in mean cell number per blastocyst obtained from oocytes matured with 5 and 6 ng/ml respect to zero Mn (IVM alone) and 2 ng/ml Mn. This study provides evidence that optimal embryo development to the blastocyst stage was partially dependent on the presence of Mn during IVM. Moreover, the availability of Mn during oocyte maturation ensures ‘normal’ intracellular GSH content in COCs and protects DNA integrity of cumulus cells.     

Components of electroencephalographic responses to slaughter in halothane-anaesthetised calves: Effects of cutting neck tissues compared with major blood vessels

AIM: To identify whether cutting neck tissues or cutting major blood vessels initiates the mechanisms responsible for electroencephalographic (EEG) responses to slaughter by ventral-neck incision without prior stunning in halothane-anaesthetised calves.

METHODS: Calves were assigned to two groups, viz transection of neck tissues with intact blood circulation through the brain (n=10), or transection of the major blood vessels of the neck but not most other neck tissues (n=7). They were minimally anaesthetised with halothane, using an established anaesthesia protocol. The animals in the neck-tissue transection group had their carotid arteries and jugular veins exposed and cannulated proximal and distal to the proposed site of subsequent ventral-neck incision; this diverted blood fl ow through these vessels so that cerebral perfusion and drainage were preserved. In animals in the blood-vessel transection group, the carotid arteries and jugular veins were exposed bilaterally by surgical dissection. They were then transected without further damage to the remaining structures of the neck. Changes in the median frequency (F50), 95% spectral edge frequency (F95), total power of the EEG (Ptot), and arterial blood pressure were compared within each group before and after neck-tissue or blood-vessel transection, and between groups following treatments.

RESULTS: Neck-tissue transection resulted in little overall change in the F50, an increase in the F95, and an initial increase in Ptot followed by a transient decrease and eventual return to pre-treatment values. There was between-animal variation in these EEG parameters. Transection of the major blood vessels of the neck resulted in a decrease in F50 in most animals; changes in F95 were highly variable, and there was a decrease in Ptot.

CONCLUSIONS: The EEG responses seen following necktissue and blood-vessel transection were qualitatively distinct, and suggested that cutting neck tissues caused greater noxious sensory input than transection of only the major blood vessels of the neck. These observations support the conclusion that the EEG responses seen after ventral-neck incision in intact animals are primarily due to noxious stimulation as a result of incision of ventral-neck tissues and not mainly as a result of loss of blood flow through the brain.     

Retrospective study of 108 foals with septic osteomyelitis

Objective   To determine the clinical characteristics, short-term outcome and future athletic performance of foals with septic osteomyelitis.
Design   Retrospective clinical study of 108 Thoroughbred foals with radiographic evidence of bone infection that were presented at the Scone Veterinary Hospital between August 1995 and December 2001. Medical records were reviewed and information concerning signalment, the clinical, laboratory and radiographic findings, treatment and outcome was obtained. Racing records were obtained and evaluated for surviving foals that had reached racing age.
Results   Mean age of foals at initial evaluation was 39 days (range 1–180 days); 21 foals had multiple radiographic bone lesions (19.4%), and 76 had concurrent septic arthritis (70.4%). The most frequently affected bones were the femur, tibia and distal phalanx. In total, 87 foals were discharged from the hospital (80.6%), 79 survived long-term to reach racing age and 52 raced (65.8%). Overall, 48% (52/108) of the foals treated for osteomyelitis raced. Foals less than 30 days of age at the time of diagnosis, critically ill foals and those with multiple bones or joints affected were significantly less likely to be discharged from hospital. Multiple septic joints, but not multiple bone involvement, had an unfavourable prognosis for racing.
Conclusions   The prognosis for survival of foals with septic osteomyelitis or osteitis is favourable. Multiple bone or joint involvement is an important short-term prognostic indicator; however, the involvement of multiple joints, but not multiple infected bones, is associated with an unfavourable prognosis for racing.     

The effect of the time interval between exposures on the susceptibility of chickens to superinfection with Marek's disease virus

Marek's disease virus (MDV) is ubiquitous within commercial poultry flocks because current vaccines do not prevent MDV infection or transmission. In order for newly-evolved MDV strains to become established within a flock, it seems inevitable that any new strain would need to infect and replicate in chickens previously infected with resident MDV strains. This phenomenon is difficult to detect and there is no clear evidence that it is even possible. Four experiments were performed to demonstrate superinfection and evaluate the effect of time between challenges on the effect of superinfection with the use of two pairs of fully virulent MDV strains that could be discriminated by novel technology: 1) JM/102W and rMd5//38CVI, and 2) rMd5 and rMd5//38CVI. Feather follicle epithelium (FFE), spleen, and tumor samples were collected at single or multiple time points from the same bird to determine the frequency and distribution of each virus present following superinfection, with the use of pyrosequencing and immunohistochemistry. Superinfection was observed in 82 of 149 (55%) FFE samples following short-interval challenge (24 hr) compared to only 6 of 121 (5%) samples following long-interval challenge (13 days), indicating a strong influence of challenge interval. In cases where the first inoculated virus was weak or delayed, the second inoculated virus was detected in 42 of 95 (44%) birds. In tumors from dually challenged birds, the second virus was again present much more often following short-interval challenge (68%) compared to long-interval challenge (11%). Virus mixtures in tumors were less common compared to those in FFE samples. Vaccination with turkey herpesvirus had no significant effect on the virus frequency for either virus pair or challenge time interval, suggesting these conclusions may be applicable to vaccinated chickens in the field. These studies demonstrated superinfection for the first time with two fully virulent MDV strains and suggest that short-interval challenge exposure and/or weak initial exposures may be important factors leading to superinfection--a prerequisite for the establishment of a second virus strain in the population. This model system should be useful to elucidate this important phenomenon further.     

Gingival stippling in dogs: Clinical and structural characteristics

Diagnosis of periodontal disease, a frequent disorder in dogs, rests mainly upon the assessment of changes affecting the gingiva. Loss of gingival stippling, one of the early symptoms of periodontal disease in the human, could also indicate early periodontitis in the dog.The aim of this study was to determine the presence of gingival stippling in dogs, its location on alveolar processes, its association with age and inflammation, and its relation to the microstructure of the gingiva. The study concerns clinical, scanning electron and light microscopic examination of the gingiva of 48 dogs.Stippling, i.e. the granular appearance of the gingiva resembling irregularly distributed crater-like depressions of varying sizes, was present in adult but absent in young and senescent dogs. Stippling was most prominent at the level of the canine and the carnassial teeth. In contrast to the human gingiva, depressions and elevations of the stippled gingiva of the dog were not distributed uniformly. Presence of stippling indicates the presence of well developed mucosal layers and extensive interdigitation of epithelial rete pegs and papillary layer. Interaction of inelastic collagen fibres and a high hyaluronan content within papillary layer resulting in a high turgor of tissue due to the extensive water binding capacity of hyaluronic acid could also play a role in shaping of the gingival surface.The significance of gingival stippling in the diagnosis of periodontal disease in dogs is limited, as stippling occurs in healthy but also slightly inflamed attached gingiva of adult individuals.     

How many Layers has the Adventitia? – Structure of the Arterial Tunica Externa Revisited

Tunica adventitia or tunica externa is the outer layer of the blood vessel wall. It consists of connective tissue with vasa and nervi vasorum and plays a key role in vascular health. The aim of our study was to compare the wall layers beyond tunica media in arteries of different type and location. The following arteries of pig, dog and cat were processed histologically and analysed by light microscopy: aorta ascendens, arcus aortae, aorta thoracica, aorta abdominalis, arteria (a.) femoralis, a. tibialis cranialis, a. carotis communis, a. lingualis, a. basilaris, a. cerebralis media, a. testicularis and aa. jejunales. We found two layers of connective tissue outside the media: (1) a compact layer with many elastic fibres in muscular and few in elastic arteries and (2) an outer layer of loose connective tissue. The compact layer was missing in aorta ascendens, arcus aortae and intracranial vessels. Adventitial stripping removed only the loose connective tissue layer. In spite of the still present compact layer, stripped arteries were very flimsy. We suggest using the term ‘tunica externa’ for the compact connective tissue layer and ‘tunica adventitia’ for the outermost loose connective tissue layer as in other organs. The presence of the tunica externa differs between species, arteries and arterial side, as well as the removability of tunica adventitia and tunica externa by anatomical dissection.     

Heavy metals and soil microbes

The discovery in the early 1980s that soil microorganisms, and in particular the symbiotic bacteria Rhizobium, were highly sensitive to heavy metals initiated a new line of research. This has given us important insights into a range of topics: ecotoxicology, bioavailability of heavy metals, the role of soil biodiversity, and the existence of ‘keystone’ organisms. Concurrently, and particularly in Europe, the research led to new approaches to the protection of soils from pollution that take into account the many effects on soil microorganisms. To date these key findings have largely been ignored in the USA, although our results caused considerable controversy there. In the past decade there have been many advances in the ecotoxicological assessment of metals and their effects on soil organisms but major gaps in knowledge and theory remain with regard to how microorganisms are exposed and respond to metals in soils. In this brief review we emphasise the need for long-term experiments and basic research to forge this understanding and improve environmental protection policies.     

Inhibited development of trichostrongylid worms in grazing cattle

Inhibition of development of gastro-intestinal trichostrongylid worms was studied using successive groups of tracer calves and groups of continuously grazed calves over one year in the Tully area of North Queensland lowland wet tropics. The results, assessed by means of worms from these calves recovered at necropsy 3 weeks after their removal from pastures, showed inhibition in the development of Haemonchus placei and Cooperia punctata at the early fourth stage at the approach to and during the relatively dry period in the area. Inhibition was however minor and inhibited larvae formed but only a small percentage of worm burdens in both categories of calves, indicating that they were not in any way of major epidemiological importance. It was suggested that the minor nature of inhibition was due to the mild climatic conditions which could not produce appropriate conditioning treatment, or caused only mild selection pressure for inhibition in the area.     

Studies of reticuloendotheliosis virus-induced lymphomagenesis in chickens

Seventy-three percent of chickens inoculated with the chick syncytial strain of reticuloendotheliosis virus (REV) at hatching developed lymphomas by 39 weeks of age. Neonatal treatment with cyclophosphamide or surgical bursectomy at 2, 4, 8, or 12 weeks of age significantly (P less than 0.01) reduced lymphoma development. In a further experiment, surgical bursectomy of REV-infected chickens followed by intravenous inoculation of the chickens with a single cell suspension of their own bursa cells at 2, 4, 9, or 13 weeks of age resulted in lymphoid tumors in chickens treated at 9 or 13 weeks but not in chickens treated at 2 or 4 weeks of age. Furthermore, this treatment did not shorten the incubation period for lymphoma development. These findings argue very strongly that transforming target cells are primarily in the bursa of Fabricius. The data also suggest that a minimum residency of 4 weeks in the bursa is required for infected bursa cells to become transformed. Therefore, lymphomagenesis induced by REV in chickens appears similar to that induced by the avian leukosis virus group.