Viruses associated with outbreaks of equine respiratory disease in New Zealand

AIM: To identify viruses associated with respiratory disease in young horses in New Zealand.

METHODS: Nasal swabs and blood samples were collected from 45 foals or horses from five separate outbreaks of respiratory disease that occurred in New Zealand in 1996, and from 37 yearlings at the time of the annual yearling sales in January that same year. Virus isolation from nasal swabs and peripheral blood leukocytes (PBL) was undertaken and serum samples were tested for antibodies against equine herpesviruses (EHV-1, EHV-2, EHV-4 and EHV-5), equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3).

RESULTS: Viruses were isolated from 24/94 (26%) nasal swab samples and from 77/80 (96%) PBL samples collected from both healthy horses and horses showing clinical signs of respiratory disease. All isolates were identified as EHV-2, EHV-4, EHV-5 or untyped EHV. Of the horses and foals tested, 59/82 (72%) were positive for EHV-1 and/or EHV-4 serum neutralising (SN) antibody on at least one sampling occasion, 52/82 (63%) for EHV-1-specific antibody tested by enzyme-linked immunosorbent assay (ELISA), 10/80 (13%) for ERAV SN antibody, 60/80 (75%) for ERBV SN antibody, and 42/80 (53%) for haemagglutination inhibition (HI) antibody to EAdV-1. None of the 64 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. Evidence of infection with all viruses tested was detected in both healthy horses and in horses showing clinical signs of respiratory disease. Recent EHV-2 infection was associated with the development of signs of respiratory disease among yearlings [relative risk (RR)=2.67, 95% CI=1.59-4.47, p=0.017].

CONCLUSIONS: Of the equine respiratory viruses detected in horses in New Zealand during this study, EHV-2 was most likely to be associated with respiratory disease. However, factors other than viral infection are probably important in the development of clinical signs of disease.     

Hyperkalemic periodic paralysis, plasma lactate and exercise tolerance

The purpose of this, trial was to determine the effect of hyperkalemic periodic paralysis (HYPP) on exercise tolerance in Quarter Horses. Five HYPP affected and five nonaffected horses were matched for age, size, gender and reproductive status. HYPP status was diagnosed by DNA analysis and potassium chloride challenge testing. Plasma lactate concentration and heart rate were used as indicators of work intensity. Serum potassium concentrations were also monitored. Two exercise experiments were conducted, the first being forty-five minutes of slow, aerobic exercise (hacking) and the other being moderate, partially anaerobic exercise (galloping). Post-exercise the horses were cooled out by randomly assigning them to either forty minutes, of standing still or forty minutes of walking. Heart rates of HYPP affected and unaffected horses were not significantly different during exercise or recovery. Plasma lactate concentrations changed slightly following slow exercise and were significantly higher for HYPP affected horses (P=0.01).At the end of exercise, values were 1.4±0.2 mmol/L and 1.0±0.1 mmol/L for HYPP affected and unaffected horses, respectively. Following moderate exercise, plasma lactate concentrations were much greater, and the difference (P<0.001) between affected and unaffected horses was more marked: immediately following exercise concentrations were 10.6±1.8 and 6.2±1.0 mmol/L in affected and unaffected horses, respectively. The higher post-exercise plasma lactate concentrations in affected horses indicates increased anaerobic muscle metabolism. Serum potassium concentrations rose following exercise and significantly higher values were seen in horses that were walked rather than stood still post-exercise.Hyperkalemic periodic paralysis (HYPP) is a dominant autosomal genetic defect occurring in American Quarter Horses and related breeds. ^1–5 The condition is widely geographically distributed and has been estimated to affect 0.4% of all Quarter Horses.⁶ Signs include sporadic attacks of muscle fasciculation, muscle spasm, sweating and weakness. Oral administration of potassium chloride produces a more severe hyperkalemia in HYPP affected horses and induces clinical attacks.¹ Electromyography reveals widespread continuous, spontaneous, muscle contraction.² Some people believe that HYPP affected horses are suitable for riding⁷; others have expressed doubts about the safety of this practice and the exercise tolerance of affected horses. HYPP affected horses have an unstable muscle membrane potential causing random muscle fiber contractions,^2,8 which could antagonize purposeful movement. This, in combination with increases in extracellular potassium concentration occurring during exercise,^9–11 may cause affected horses to stumble or collapse while being ridden, posing a danger to both horse and rider. In HYPP affected people, potassium concentration rises approximately one to two hours after exercise during which time HYPP attacks can occur.^9,10 For these reasons, the exercise tolerance of HYPP affected horses and the effects of exercise on spontaneous HYPP attacks deserves investigation.We standardized the exercise test and chose heart rate and plasma lactate concentrations as indicatoors, of exercise tolerance and energy metabolism.¹² Heart rate is the major determinant of oxygen delivery to muscle and the rate rises with exercise intensity until it reaches a plateau at high velocities.^13,14 Several lines of investigation indicate that plasma lactate concentrations reflect muscle work. Lactate release from resting muscle is minimal but large amounts are released during strenuous exercise.^15–17 Lactate release increases with increasing muscle work.^16,18 During exercise, muscle oxygen consumption increases until eventually a plateau is reached after which no further increase in oxygen consumption in response to increased work loads is possible.¹⁸ At this point the mitochondrial electron chain transport system is operating at its maximal possible rate, energy production can only be supplemented by the use of inefficient anerobic glycolytic metabolism and the muscle starts to release lactate. If the concentrations of plasma lactate are high, the muscles are depending on anaerobic metabolism and are working closer to exhaustion^16,18–21The objectives of this experiment were twofold. One was to determine if HYPP adversely affects exercise performance. The second was to study the effects of cooling out, by either standing still or walking, on serum potassium and plasma lactate concentrations.     

VASA (DDX4) is a Putative Marker for Spermatogonia,Spermatocytes and Round Spermatids in Stallions

Expression of the protein DDX4/MVH, or VASA, has been reported in germ cells of several species. The main objectives of this study were to (i) investigate VASA expression patterns in testicular cells of stallions at two different reproductive stages (pre‐pubertal and post‐pubertal) and (ii) evaluate the use of VASA antibody as a molecular marker for single germ cells from stallions. Testicular tissues were obtained from stallions and categorized as pre‐pubertal and post‐pubertal based on the formation of lumen and status of spermatogenesis on the cross section of seminiferous tubules. The results of Western blot showed a VASA protein band located at 76 kDa, indicating that the rabbit antibody has a cross‐reactivity with horse testicular tissues. The result of immunolabelling showed that VASA was expressed in the cytoplasm of spermatogonia at both reproductive stages and in spermatocytes and round spermatids at the post‐pubertal stage. GATA4‐positive Sertoli cells and Leydig cells located in the interstitial space were not immunolabelled with VASA. These results suggest that VASA can be utilized as a molecular marker for germ cells of stallions at pre‐pubertal and post‐pubertal stages. Interestingly, immunolabelling intensity was significantly higher in pachytene spermatocytes compared to spermatogonia and round spermatid. VASA antibody was also effective for staining of single germ cell preparations. In conclusion, VASA protein expression can be used as a marker for identification of spermatogonia, spermatocytes and round spermatids in testicular tissues of stallions.     

Widespread iron limitation of phytoplankton in the south pacific ocean

Diel fluorescence patterns were discovered in phytoplankton sampled over 7000 kilometers of the South Pacific Ocean that appear indicative of iron-limiting growth conditions. These patterns were rapidly lost after in situ iron enrichment and were not observed during a 15,000-kilometer transect in the Atlantic Ocean where iron concentrations are relatively high. Laboratory studies of marine Synechococcus sp. indicated that the patterns in the South Pacific are a unique manifestation of iron limitation on the fluorescence signature of state transitions. Results suggest that primary productivity is iron limited not only throughout the equatorial Pacific but also over much of the vast South Pacific gyre.     

Clustering of leukocyte and fibroblast interferon genes of human chromosome 9

At least ten leukocyte interferon genes and the single known fibroblast interferon gene have been localized on the pter leads to q12 region of human chromosome 9. Gene mapping was accomplished by blot hybridization of cloned interferon complementary DNA to DNA from human-mouse cell hybrids with a translocation involving human chromosome 9. Supporting evidence suggests these genes are clustered.     

Integration of environmental, agronomic, and economic aspects of fertilizer management

Nitrogen fertilization is a substantial source of nitrogen-containing trace gases that have both regional and global consequences. In the intensive wheat systems of Mexico, typical fertilization practices lead to extremely high fluxes of nitrous oxide (N2O) and nitric oxide (NO). In experiments, lower rates of nitrogen fertilizer, applied later in the crop cycle, reduced the loss of nitrogen without affecting yield and grain quality. Economic analyses projected this alternative practice to save 12 to 17 percent of after-tax profits. A knowledge-intensive approach to fertilizer management can substitute for higher levels of inputs, saving farmers money and reducing environmental costs.     

Myotonic dystrophy type 2 caused by a CCTG expansion in intron 1 of ZNF9

Myotonic dystrophy (DM), the most common form of muscular dystrophy in adults, can be caused by a mutation on either chromosome 19q13 (DM1) or 3q21 (DM2/PROMM). DM1 is caused by a CTG expansion in the 3' untranslated region of the dystrophia myotonica-protein kinase gene (DMPK). Several mechanisms have been invoked to explain how this mutation, which does not alter the protein-coding portion of a gene, causes the specific constellation of clinical features characteristic of DM. We now report that DM2 is caused by a CCTG expansion (mean approximately 5000 repeats) located in intron 1 of the zinc finger protein 9 (ZNF9) gene. Parallels between these mutations indicate that microsatellite expansions in RNA can be pathogenic and cause the multisystemic features of DM1 and DM2.