Separation of DNA for molecular markers analysis from leaves of the Vitis vinifera

In the present study, three DNA extraction procedures were examined to determine which might yield DNA from Grape leaves suitable for molecular analysis for RAPD, SSR. AFLP and etc analysis. The three methods examined were: the miniprep procedure and the modified CTAB for difficult species and protocol CTAB. Only the modified CTAB method consistently yielded DNA suitable for Polymerase Chain Reaction (PCR) amplification, regardless of plant growing conditions or leaf age. The quality and quantity of extracted genomic DNA gained from these methods are deliberated by means UV biophotometer, electrophoresis in 1.2% agarose gel and PCR. In this regard, application chosen for young and mature leaves, the most value of qualified DNA, is extracted from fully expanded leave when PVP was added to the extraction buffer. This same procedure also yielded PCR-amplifiable DNA from various other perennial, woody species and from other fruit species such as apple (Malus domestica), cherry (Prunus avium), peach (Prunuspersica), plum (Prunus domestica). DNA yield from this procedure is high (up to 1 mg g(-1) of leaf tissue). DNA is completely digestible with restriction endonucleases and amplifiable in the Polymerase Chain Reaction (PCR).     

Improved feed intake and body weight change in sheep treated with dexamethasone at entry into pens or feedlots

Treatment with the synthetic glucocorticoid dexamethasone sodium phosphate increased the feed intake of sheep within 24 h. A single treatment with the longer-acting compound dexamethasone trimethylacetate had a slower effect, but resulted in improved feed intake and enhanced body weight gain over a period of 7 to 14 days. Treatment was effective in sheep housed individually and in sheep penned in groups. The improvement in body weight gain disappeared 15 to 20 days after a single treatment, but in most studies treated sheep had less variability in weight change than untreated sheep at this time. A statistically significant increase in body weight gain was observed in 8 of 11 trials; in the other 3 trials, it appeared that the feed intake of the untreated sheep was already maximal. Wethers treated with dexamethasone trimethylacetate on arrival at an assembly point for live export arrived in the Middle East 23 days later with a lower mean weight loss, partly because fewer sheep lost a significant amount of weight.     

Effect of bandaging on second intention healing of wounds of the distal limb in horses

Objective   To evaluate the effect of a non-occlusive dressing incorporated in a 3-layer bandage on second intention healing of wounds of the distal portion of the limb.
Study Design   Untreated wounds in 33 adult horses used in four studies using the same wound-healing model conducted over 5 years.
Methods   Standardised, full-thickness wounds were made in the skin overlying the dorsomedial aspect of the mid-metacarpus; 17 horses were bandaged with a non-occlusive dressing covered by gauze-coated cotton wool that was compressed with adhesive tape; 16 horses were left unbandaged. Wounds were photographed weekly for 9 weeks and the images were analysed electronically.
Results   There were significant effects associated with bandage (P < 0.0001), week (P < 0.001), and bandage by week interaction (P < 0.0001). There was no difference in wound area at the first time-point after wound creation (P = 0.38). After week 1, there was a difference between bandaged and unbandaged wounds in wound area at each measurement until the end of the study. Bandaged wounds showed greater and more prolonged retraction. Unbandaged wounds retracted for 2 weeks before beginning to contract, whereas bandaged wounds continued to retract for 3 weeks. In bandaged wounds excess granulation tissue required regular trimming, but not in unbandaged wounds. There was no difference between groups in the total days to healing or the overall rate of healing.
Conclusions   These results should be treated with caution until validated with contemporaneous, controlled studies. Covering a wound with a non-occlusive dressing in a 3-layer bandage led to greater wound retraction, modulated the rate of wound contraction and promoted excessive granulation tissue. If excessive granulation tissue is excised regularly, bandaging has no effect on total time to healing.     

Involvement of PAPP‐A and IGFR1 in Cystic Ovarian Disease in Cattle

Cystic ovarian disease (COD) is one of the main causes of infertility in dairy cattle. It has been shown that intra‐ovarian factors, such as members of the insulin‐like growth factor (IGF) system, may contribute to follicular persistence. The bioavailability of IGF to initiate its response by binding to specific receptors (IGFRs) depends on interactions with related compounds, such as pregnancy‐associated plasma protein A (PAPP‐A). The aim of this study was to determine IGFR1 and PAPP‐A expression both in follicles at different stages of development and in cysts, to evaluate the roles in the etiopathogenesis of COD in cattle. The mRNA expression of PAPP‐A was higher in granulosa cells of large tertiary follicles than in cysts, whereas the protein PAPP‐A present in the follicular fluid from these follicles showed no differences. Although no PAPP‐A mRNA expression was detected in smaller tertiary follicles, in their follicular fluid, this protease was detected in lesser concentration than in cysts. The mRNA expression of IGFR1 was lower in granulosa cells from cystic follicles than in those from tertiary ones. However, the protein expression of this receptor presented the highest levels in cystic structures, probably to increase the possibility of IGF response. The data obtained would indicate that animals with COD have an altered regulation of the IGF system in the ovary, which could be involved in the pathogenesis of this disease in cattle.     

Profiling the New Zealand Thoroughbred racing industry. 2. Conditions interfering with training and racing

AIM: To describe and enumerate conditions that interrupted training and racing in a population of Thoroughbred racehorses in New Zealand.

METHODS: A longitudinal study design was used to collect data on horses training under the care of 20 licensed racehorse trainers from venues in the mid to lower regions of the North Island between October 1997 and July 2000. Incidence rates were reported for first and second occurrences for different categories of musculoskeletal injury (MSI), and first occurrences of upper and lower respiratory tract disease, using training days as time-at-risk. The proportion of horses that retired or died due to MSI, respiratory tract or miscellaneous conditions was used to estimate risk of exit for each type of event. Duration of training preparation, starts per 100 training days, and proportion of starts that ended in first, second or third place, were calculated for horses at risk for first MSI, and all subsequent MSIs. In training preparations that had at least one start and that ended in MSI, the cumulative percentage of MSIs by day of diagnosis was reported for 0–21 days after the last start in the preparation.

RESULTS: Horses (n=1,571) were followed during 3,333 training preparations and 392,290 training days. Events associated with the end of a training preparation or spell period included MSI (n=834), respiratory event (RE; n=165), miscellaneous event (ME; n=58), and voluntary retirements (n=360). Causes of MSI included lameness (n=400), shin soreness (n=207), tendon and ligament conditions (n=98), injury or laceration (n=56), fractures (n=55), and back disorders (n=18). MSIs involved the limbs in 97% of cases, and the lower limbs up to the carpus or hock in the fore- and hindlimbs, respectively, in 81% of cases. Most (93%) lower limb conditions involved a forelimb while 70% of MSIs that involved structures above the carpus or hock involved a hindlimb. Incidence rates (IRs) are reported for each age group for first and second occurrences of MSI, and first occurrence of upper and lower respiratory tract disease. The risk of MSI was higher in horses that had incurred one previous MSI (RR 1.4, 95% CI=1.2–1.7; p>0.001) than in horses without any previous MSI. The proportion of horses that exited due to death or retirement varied with the type of injury, and the highest proportion was associated with recurrent fractures, and tendon and ligament injuries (46.2 and 44.4%, respectively). The overall IR of horses exiting the study due to retirement or death increased with increasing age, and was higher in females than males for horses aged 2, 3, 4, and ≥5 years. A reduction in the number of starts per 100 training days was observed in horses aged ≥5 years when returning to training after an initial MSI (p=0.004). Male horses of all age groups and females younger than 4 years had shorter median training preparations (p>0.05) when returning to training after an initial MSI compared with preparations at risk for a first occurrence of MSI. Between 27 and 62% of cases of MSI that occurred in training preparations after at least one start were reported on the day of the last start, and the remainder were reported in the days to weeks following the last start of that preparation.

CONCLUSION: Incidence rates, and proportions of affected horses that retired or died as a result of injury or disorder varied with type of injury and age of horse. Horses returning to training after an initial MSI were at higher risk of subsequent MSIs and showed changes in duration of training preparations, but little change in starts per 100 training days or probability of placing in each start. MSIs in racing horses were less likely to be reported on the day of a race than at other times in the training preparation for all ages except 2-year-olds.     

Effect of training location and time period on racehorse performance in New Zealand. 2. Multivariable analysis

AIM: To investigate training location (horses trained in Matamata vs those trained at all other venues in New Zealand), and time period (1996–1997 and 1998–1999), while controlling for other horse- and race- or trial-related factors, as a means of assessing the possible impact of construction of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance (number of starts, and failure to race within 6 months of any start).

METHODS: Multivariable logistic regression and poisson analysis were used to analyse data derived using a retrospective cohort approach. Multivariable logistic regression was also used to analyse a case-control study. All data were derived from New Zealand Thoroughbred Racing (NZTR), records of race and trial results for racehorses trained in Matamata and other venues in New Zealand, covering two 19-month time periods (1996–1997 and 1998–1999). Outcome variables included whether a horse started again in the 6 months following any start that occurred in the first 13 months of either time period, and a count of the total starts for every horse.

RESULTS: Factors associated with increased risk of a start being followed by a 6-month no-race period included training location other than Matamata in comparison to horses trained in Matamata in the 1996–1997 time period, increasing age, 1998–1999 over 1996–1997, starting in a trial rather than a race, placing fourth or worse in a start, softer track conditions, summer vs autumn, increasing cumulative exercise intensity in the 60 days prior to a start, and increasing race distance. Factors associated with an increase in the total number of starts included horses trained at Matamata in 1996–1997 compared with other time period-location combinations, younger age of horses at the time of a start, longer race distance, and an increasing proportion of starts in stakes races.

CONCLUSIONS: Official race and trial results data provided a valuable resource for epidemiological studies of factors influencing racehorse performance. Results of analyses performed here provided little evidence of any adverse impact of a new training surface at the Matamata Racing Club on indirect measures of racehorse performance.     

Permanent infertility in ewes exposed to plant oestrogens

In Australia, more than 1 million ewes have permanently damaged reproductive tracts because they have been grazed on oestrogenic pasture. These effects occur in the absence of classical clinical "clover disease". The lesions result from an "organisational" action of oestrogen, causing a mild sexual transdifferentiation to occur in ewes during adult life, with the main lesion being found in the cervix. Diagnosis of the problem depends primarily on detection of the pathological changes, rather than history or clinical signs. On average, affected flocks have around a 10% increase in non-pregnant ewes, but most farmers can achieve an acceptable lambing rate by increasing their management and feed inputs to produce more twin lambs. Agronomic approaches have been used in the past to combat "clover disease", but further progress with the present problem may depend on the development of animal-based solutions.     

The large‐scale capture of eastern grey kangaroos (Macropus giganteus) and red kangaroos (Osphranter rufus) and its application to a population management project

Objective: A large‐scale capture method was developed to enable sterilisation of a macropod population in western Sydney from 2005 to 2018. Methods: Until March 2007, free ranging eastern grey kangaroos and red kangaroos were herded into purpose‐built 15 m diameter capture yards (CYs) for darting with a projectile syringe. From March 2007 onwards, animals were free‐range darted in large areas without herding. Kangaroos were darted with 1.33–5.10 mg/kg tiletamine/zolazepam and 0.01–0.02 mg/kg medetomidine, ± 0.03 mg/kg acepromazine. Deaths were monitored. Population counts were performed annually. Results: There were 5825 capture events involving 3963 kangaroos. Over 85% of all captures occurred from 2005 to 2008. Of all reported deaths (n = 523), 135 were attributed to ill health. Musculoskeletal injuries incurred during capture were the main project‐related cause of death (n = 116). Post capture myopathy was uncommonly diagnosed following capture (n = 19). Conclusion: The herding and capture method enabled a large number of kangaroos to be mobilised and captured with low mortality rates, and the use of CYs resulted in fewer capture‐related injuries and deaths than free‐range capture. The drug doses and combinations used for darting were safe and effective, and the capture technique was successfully applied to a population management project.     

Classification of soils supporting mangrove plantation in Kuwait

The State of Kuwait has initiated an ambitious national program to establish greenery in 20000 ha of open spaces in inland and coastal areas. Afforestation of intertidal zones with mangrove plants is being considered as a viable option for greening coastal areas. In view of this fact, experimental plantations were established using two ecotypes of Avicennia marina (Bahrain and UAE) and one ecotype of Avicennia germinans (Florida). The study consisted of two 10-year old plantations and two 2-year old plantations. Seedlings established successfully and grew vigorously on some of the locations within the site. Therefore, studies were conducted during 2002-2003 to describe and classify soils that supported maximum mangrove establishment and growth under the harsh arid coastal conditions of Kuwait. For this purpose, eight soil profiles (four each representing areas where seedlings established successfully and grew vigorously and where all seedlings died in the initial stages of establishment) were investigated and classified according to the soil taxonomy. The main soil types observed in the study site were: Typic Aquisalids, Typic/sodic Aquicambids and Calcic Aquisalids. Mangrove growth performance in relation to the landform, soil type and properties of the soil at various depths is discussed in the paper. The plant growth appeared to be the better on the typic Aquisalid soil than that on the Typic/sodic Aquicambid soils. The relative proportions of the coarse sand fraction, occurrence of the anaerobic layer in the profile and the surface layer salinity influenced both the establishment and growth of mangrove plants.