Retinopathy in Greyhound dogs: Prevalence, fundoscopic, and histopathological findings.

OBJECTIVES: To determine the prevalence of retinal lesions and describe the fundoscopic findings of retinopathy in Greyhound dogs in the Manawatu/Whanganui region of New Zealand. To examine possible associations between sex, age, and racing variables with retinopathy in the study population. To describe retinal histologic findings in seven Greyhounds with retinopathy in New Zealand. METHODS: Two hundred Greyhound dogs from the Manawatu/Whanganui region of New Zealand underwent fundoscopy and fundic photography to identify and score the degree of retinopathy. Associations between retinopathy and age, sex, as well as racing variables, were examined. Histologic examination of the retina was undertaken on the eyes of seven Greyhounds from the Manawatu and Canterbury regions previously diagnosed with retinopathy by fundoscopy. RESULTS: Fifty dogs (25.1%) were identified with retinopathy of varying degrees of severity. In at least one eye, 7.5% of dogs had mild retinopathy, 11.6% moderate retinopathy, and 6.0% severe retinopathy. Males were more likely to be affected in both eyes and with moderate or severe grades, than females. Increasing age was not associated with increased prevalence of retinopathy, nor increased grade of severity. Retinal histology identified multifocal retinal detachment in 5 of the 7 cases examined and other common lesions included choroidal necrosis and outer to full-thickness retinal atrophy in the absence of significant inflammation. CONCLUSIONS: Retinopathy is prevalent in Greyhounds in the Manawatu/Whanganui region of New Zealand, but more research is required to elucidate the etiopathogenesis. Consideration should be made to include mandatory eye health examination in racing Greyhound dogs.

Stepping behavior and muscle activity of dairy cattle standing on concrete or rubber flooring for 1 or 3 hours.

The type of flooring in dairy cattle systems influences cows' health and welfare. Although concrete is common, the use of more compressible flooring, such as rubber, is increasing. Cows prefer to stand and walk on rubber surfaces than on concrete; however, it is largely unknown how walking and standing for longer periods of time influence muscle activity and fatigue. Therefore, we used measures of behavior and muscle activity to investigate the potential benefits of providing a rubber flooring surface to dairy cattle. Sixteen lactating Holstein cows were forced to stand on either concrete or rubber flooring for 1 or 3 h in a 2 × 2 crossover design. Surface electromyograms (SEMG) and skin surface temperature were used to evaluate muscle activity, fatigue, and movement of muscle activity between the hind legs. Activity of 2 muscles, the bicep femoris and middle gluteal, was assessed during both static contractions, when cows transferred weight to each hind leg, before and after 1 and 3 h of standing, and dynamic contractions, associated with steps and with shifts in weight without steps. In addition, we evaluated stepping rate, time between each step, feeding behavior, skin surface temperature, and latency to lie down after standing. Standing duration influenced both the behavior and muscle activity of cows. Stepping rate increased with standing time for cows on both flooring types. Static muscle activity parameters of the bicep femoris muscle were higher after 3 h of standing for cows standing on both flooring types (2.3 and 3.6% increases in median amplitude and median power frequency, respectively) compared with the change after 1 h and compared with baseline values before the standing treatment. Flooring type influenced the behavior and muscle activity of the cows, particularly during the first hour of exposure; cows standing on rubber had a higher stepping rate, shorter interval between steps, and higher number of SEMG shifts (muscle activity shifts with or without visible steps) than cows on concrete. There was no difference in skin surface temperature, feeding behavior, or latency to lie down between the treatments. The results show that standing on a rubber flooring caused a different initial behavioral response compared with standing on concrete; however, possible reasons for these changes are unclear. Standing for 3 h resulted in an increase in stepping rate and in some muscle activity parameters; however, the results regarding muscle fatigue in relation to flooring type are inconclusive.

Aploneura lentisci (Homoptera: Aphididae) and Its Interactions with Fungal Endophytes in Perennial Ryegrass (Lolium perenne).

Aploneura lentisci Pass. is endemic to the Mediterranean region where it is holocyclic, forming galls on its primary host, Pistacia lentiscus and alternating over a 2-year period between Pistacia and secondary hosts, principally species of Gramineae. This aphid is widely distributed in Australia and New Zealand on the roots of the common forage grasses, ryegrass (Lolium spp.) and tall fescue (Schedonorus phoenix) where it exists as permanent, anholocyclic, parthenogenetic populations. Previous studies have indicated that infestations of A. lentisci significantly reduce plant growth and may account for differences in field performance of Lolium perenne infected with different strains of the fungal endophyte Epichloë festucae var. lolii. These obligate biotrophs protect their host grasses from herbivory via the production of alkaloids. To confirm the hypothesis that growth of L. perenne is associated with the effect of different endophyte strains on aphid populations, herbage and root growth were measured over time in two pot trials that compared three fungal endophyte strains with an endophyte-free control. In both pot trials, aphid numbers were lowest on plants infected with endophyte strain AR37 at all sampling times. In plants infected with a common toxic strain naturalized in New Zealand, aphid numbers overall were lower than on uninfected plants or those infected with strain AR1, but numbers did not always differ significantly from these treatments. Populations on AR1-infected plants were occasionally significantly higher than those on endophyte-free. Cumulative foliar growth was reduced in AR1 and Nil treatments relative to AR37 in association with population differences of A. lentisci in both trials and root dry weight was reduced in one trial. In four Petri dish experiments survival of A. lentisci on plants infected with AR37 declined to low levels after an initial phase of up to 19 days during which time aphids fed and populations were similar to those on plants without endophyte. Aphids on AR37-infected plants became uncoordinated in their movement and developed tremors before dying suggesting a neurotoxin was responsible for their mortality. Results support the hypothesis that differences in A. lentisci populations due to endophyte infection status and strain affects plant growth.

Effects of space allowance on the behavior and physiology of cattle temporarily managed on rubber mats.

Dairy cattle managed in some pasture-based systems, such as in New Zealand, are predominantly kept outdoors all year around but are taken off pasture for periods, especially in wet weather to avoid soil damage. The use of rubber matting for such stand-off practices is becoming more common to improve animal welfare, and our objective was to investigate the effects of different space allowances on cow behavior and physiology when managed temporarily on rubber mats during a weather-induced stand-off period. Thirty pregnant, nonlactating Holstein-Friesian dairy cows were divided into 6 groups of 5 and exposed to 6 treatments following a Williams designed 6×6 Latin square. The treatments consisted of 6 space allowances on a 24-mm rubber surface during a simulated weather-induced stand-off period: 3.0, 4.5, 6.0, 7.5, 9.0, and 10.5 m(2)/cow. The stand-off period consisted of 18 h in the treatment pens followed by 6 h at pasture to allow for their daily feed intake (no feed was available during stand-off, following normal farm practice), for 3 consecutive days, with 6 d of recovery on pasture between treatments. When cows had more space available during the stand-off period, they spent more time lying on the rubber mats and less time lying on pasture during their daily 6-h feed break. Mean lying times (24 h, pasture and rubber mats combined) for the different space allowances were for 3.0 m(2)=7.5 h, 4.5 m(2)=10.2 h, 6.0 m(2)=11.9 h, 7.5 m(2)=12.4 h, and 10.5 m(2)=13.8 h. At 6.0 m(2) of space allowance per cow, the animals spent similar times lying per 24 h as when the cows were on recovery on pasture in between treatments (11.9 and 11.2 h, respectively). Aggressive interactions and nonaggressive lying disturbances were more frequent at lower space allowances (aggressive interactions decreased by 35% from 3.0 to 4.5 m(2)/cow, with a slower decline thereafter). Cows were dirtier after the stand-off period, particularly at lower space allowances. All cows had higher gait scores after the stand-off period; however, this change was unaffected by space allowance and very minor. Stride length, plasma cortisol, and body weight were all unaffected by the stand-off period and space allowance. The results suggest that to reduce aggressive behavior and maintain adequate lying times, dairy cattle managed temporarily on rubber matting for up to 18 h per day, without feed, should have a space allowance of at least 4.5 to 6.0 m(2) per cow.

Effects of acetyl-L-carnitine on lamb oocyte blastocyst rate, ultrastructure, and mitochondrial DNA copy number.

Viable lambs can be produced after transfer of in vitro-derived embryos from oocytes harvested from prepubertal lambs. However, this occurs at a much lower efficiency than from adult ewe oocyte donors. The reduced competence of prepubertal oocytes is believed to be due, at least in part, to deficiencies in cytoplasmic maturation. Differences in the cytoplasmic ultrastructure between prepubertal and adult oocytes have been described in the sheep, pig, and cow. Prepubertal lamb oocytes have been shown to have a different distribution of mitochondria and lipid droplets, and less mitochondria and storage vesicles than their adult counterparts. L-carnitine plays a role in supplying energy to the cell by transporting long-chain fatty acids into mitochondria for ß-oxidation to produce ATP. Both L-carnitine and its derivative acetyl-L-carnitine have been reported to increase the blastocyst rate of oocytes from mice, cows, and pigs, treated during IVM. L-carnitine has also been shown to increase mitochondrial biogenesis in adipose cells. Therefore, the aims of this study were to determine if treatment of oocytes from prepubertal lambs with acetyl-L-carnitine during IVM could increase the blastocyst rate and alter mitochondria, vesicle, or lipid droplet number, volume, or distribution. The blastocyst rate was doubled in prepubertal lamb oocytes treated with acetyl-L-carnitine when compared to untreated oocytes (10.0% and 4.6%, respectively; P = 0.028). Light microscopy, scanning electron microscopy, and stereology techniques were used to quantify organelles in untreated and acetyl-L-carnitine-treated lamb oocytes, and quantitative polymerase chain reaction methods were used to measure the mitochondrial DNA copy number. There were no differences in mitochondrial volume, number, or mitochondrial DNA copy number. Acetyl-L-carnitine treatment increased the cytoplasmic volume (P = 0.015) of the oocytes, and there were trends toward an increase in the vesicle volume (P = 0.089) and an altered distribution of lipid droplets (P = 0.076). In conclusion, acetyl-L-carnitine can be used to increase the in vitro blastocyst rate of juvenile oocytes and therefore to improve juvenile in vitro embryo transfer methods. These methods can be used for livestock improvement by increasing the rate of genetic gain. Further work is required to identify the contents of the vesicles and confirm the mode of action of acetyl-L-carnitine in improving oocyte competence.

Mitochondria and vesicles differ between adult and prepubertal sheep oocytes during IVM.

Oocytes from prepubertal animals have a reduced ability to undergo normal embryo development and produce viable offspring. The correct quantity, activity and cytoplasmic distribution of oocyte organelles are essential for oocyte maturation, fertilisation and subsequent embryo development. The aim of this study was to quantify the ultrastructural differences between oocytes from prepubertal lamb and adult ewes using electron microscopy and stereology. We also determined whether quantitative polymerase chain reaction (qPCR) methods give comparable estimates of mitochondrial number to stereology. Mean storage vesicle volume was greater in adult compared with lamb oocytes before IVM and decreased during maturation in both adult and lamb oocytes. Mitochondrial volume and number increased in adult oocytes during maturation; however, no increase was observed in lamb oocytes. Mitochondrial DNA copy number measured by qPCR showed no differences between adult and lamb oocytes. A different distribution of mitochondria was observed in lamb oocytes before maturation, while the percentage of hooded mitochondria increased during maturation in adult oocytes and decreased in the lamb. In conclusion, the present study has identified differences in the vesicles and mitochondria between adult and lamb oocytes from ewes that may contribute to reduced developmental competence in prepubertal oocytes.

A structural basis for the reduced toxicity of dinophysistoxin-2.

Okadaic acid (OA), dinophysistoxin-1 (DTX-1), and dinophysistoxin-2 (DTX-2) are algal toxins that can accumulate in shellfish and cause diarrhetic shellfish poisoning. Recent studies indicate that DTX-2 is about half as toxic and has about half the affinity for protein phosphatase 2A (PP2A) as OA. NMR structural studies showed that DTX-1 possessed an equatorial 35-methyl group but that DTX-2 had an axial 35-methyl group. Molecular modeling studies indicated that an axial 35-methyl could exhibit unfavorable interactions in the PP2A binding site, and this has been proposed as the reason for the reduced toxicity of DTX-2. Statistical analyses of published data indicate that the affinity of PP2A for DTX-1 is 1.6-fold higher, and for DTX-2 is 2-fold lower, than for OA. We obtained X-ray crystal structures of DTX-1 and DTX-2 bound to PP2A. The crystal structures independently confirm the C-35 stereochemistries determined in the earlier NMR study. The structure for the DTX-1 complex was virtually identical to that of the OA-PP2A complex, except for the presence of the equatorial 35-methyl on the ligand. The favorable placement of the equatorial 35-methyl group of DTX-1 against the aromatic pi-bonds of His191 may account for the increased affinity of PP2A toward DTX-1. In contrast, the axial 35-methyl of DTX-2 caused the side chain of His191 to rotate 140 degrees so that it pointed toward the solvent, thereby opening one end of the hydrophobic binding cage. This rearrangement to accommodate the unfavorable interaction from the axial 35-methyl of DTX-2 reduces the binding energy and appears to be responsible for the reduced affinity of PP2A for DTX-2. These results highlight the potential of molecular modeling studies for understanding the relative toxicity of analogues once the binding site at the molecular target has been properly characterized.