Developments in methodology of plant virus detection

As a general rule some form of polyclonal-antibody based enzyme immunoassay is still preferred for routine plant virus detection, but modifications may be necessary when increased sensitivity or specificity is required. In recent years new developments in antibody-based detection methods have mostly involved the provision of specific reagents, such as monoclonal antibodies or affinity-purified second antibody-enzyme conjugates which have helped to improve the sensitivity and specificity of the assays. Other kinds of tests (such as nucleic acid hybridisation) must be used when tests for virus coat protein are not appropriate. Recently, tests based on the polymerase chain reaction show great promise. There is no single universal test: assays must be devised and optimised for each pathogen. The challenge with all types of test is to make them quicker, less labour intensive, and —if possible — cheaper.     

In vitro Effect of Zearalenone and α-Zearalenol on Boar Sperm Characteristics and Acrosome Reaction

This study was conducted to determine the in vitro effects of three different concentrations (125, 187.5 and 250 microM in diluted semen) of zearalenone (zen) and alpha-zearalenol (alpha-zen) on boar sperm. Semen parameters such as motility, viability and spontaneous acrosome reaction were evaluated. From the results it was shown that both zen and alpha-zen affected the sperm characteristics significantly (p < 0.05), except for alpha-zen at the low concentration which did not decrease the percentage of live reacted spermatozoa significantly. In conclusion, zen and alpha-zen are directly toxic when they affect boar semen in vitro and consequently decrease the fertilization ability of the sperm. The higher the concentration of mycotoxin tested, the greater the decline of sperm parameters noticed. The influence of mycotoxins was found to be time- and dose-dependent.     

Hyponatremia and hyperkalemia associated with idiopathic or experimentally induced chylothorax in four dogs

Two dogs with idiopathic chylothorax and 2 dogs with experimentally induced (ie, ligation of the cranial vena cava) chylothorax were treated by intermittent thoracic drainage. Of these 4 dogs, 3 that did not have evidence of renal failure had normal or near-normal serum sodium and potassium concentrations before thoracic drainage began, and all 3 developed repeatedly marked hyponatremia and hyperkalemia during thoracic drainage. Another dog became weak and depressed, ostensibly because of hyperkalemia. Serum sodium and potassium concentrations in 1 dog with spontaneous chylothorax returned to normal after chylothorax resolved and thoracic drainage was stopped. The other 3 dogs died or were euthanatized, and the effect of stopping thoracic drainage could not be evaluated. In 3 dogs in which it was measured, normal-to-high plasma cortisol concentration was observed before and after adrenocorticotropin administration, and 2 dogs also had hyperaldosteronemia. Hyponatremia was hypothesized to be caused by sodium loss via thoracic drainage whereas hyperkalemia may have been multifactorial in origin, but probably was attributable, at least, in part to decreased renal potassium clearance.     

Does Clinical Treatment with Phenylbutazone and Meloxicam in the Pre‐ovulatory Period Influence the Ovulation Rate in Mares?

The presence of anovulatory haemorrhagic follicles during the oestrous cycle of mares causes financial impacts, slowing conception and increasing the number of services per pregnancy. Non‐steroidal anti‐inflammatory drugs (NSAIDs) such as meloxicam and phenylbutazone are used in the treatment of several disorders in mares, and these drugs can impair the formation of prostaglandins (PGs) and consequently interfere with reproductive activity. This study aimed to evaluate the effects of treatment with NSAIDs on the development of pre‐ovulatory follicles in mares. In total, 11 mares were studied over three consecutive oestrous cycles, and gynaecological and ultrasound examinations were performed every 12 h. When 32‐mm‐diameter follicles were detected, 1 mg of deslorelin was administered to induce ovulation. The first cycle was used as a control, and the mares received only a dose of deslorelin. In the subsequent cycles, in addition to receiving the same dose of deslorelin, each mare was treated with NSAIDs. In the second cycle, 4.4 mg/kg of phenylbutazone was administered, and in the third cycle, 0.6 mg/kg of meloxicam was administered once a day until ovulation or the beginning of follicular haemorrhage. All of the mares ovulated between 36 and 48 h after the induction in the control cycle. In the meloxicam cycle, 10 mares (92%) did not ovulate, while in the phenylbutazone cycle, nine mares (83%) did not ovulate. In both treatments, intrafollicular hyperechoic spots indicative of haemorrhagic follicles were observed on ultrasound. Thus, our results suggested that treatment with meloxicam and phenylbutazone at therapeutic doses induced intrafollicular haemorrhage and luteinization of anovulatory follicles.     

Testing Usability of Butylated Hydroxytoluene in Conservation of Goat Semen

The objective of this study was to investigate whether butylated hydroxytoluene (BHT) could be used as a suitable supporter or alternative of egg yolk during preservation of goat spermatozoa. Three in vitro experiments and a fertility test were conducted to evaluate the effect of BHT on viability of chilled‐stored semen as well as motility and kidding rate of frozen‐thawed spermatozoa. In the first two experiments, ejaculates (n = 30/experiment) were collected from 10 bucks, split, diluted with egg yolk‐based and egg yolk‐free extenders supplemented with or without 0.3, 0.6, 2, 5 and 8 mm BHT and stored at 5°C for 168 h. In the third experiment, 30 ejaculates were collected from the above‐mentioned bucks, split and diluted with egg yolk‐free extenders supplemented with or without 0.3, 0.6 and 0.9 mm BHT and egg yolk‐based extenders supplemented with or without 5 mm BHT. Diluted semen was cooled to 5°C over a period of 4 h, frozen and thawed in the form of 0.3‐ml pellets. In the fertility test, 75 ejaculates were collected from two proven fertile bucks, split, diluted with egg yolk‐free extenders containing 0.6 mm BHT and egg yolk‐based extenders supplemented with or without 5 mm BHT, frozen and thawed as described above. An insemination volume of 0.6 ml containing 120–140 × 10⁶ progressively motile spermatozoa was used for a single cervical insemination of cloprostenol‐synchronized does (n = 230). The results showed that addition of 5 mm BHT to egg yolk‐deficient (2.5%) extenders significantly improved viability of chilled‐stored semen together with motility (48.5%) and fertility (62.5%) of frozen‐thawed spermatozoa. Replacement of egg yolk in semen extenders by 0.6 mm BHT could sustain not only viability of chilled‐stored semen but also post‐thaw motility (47.5%) and fertility (53.75%) of frozen‐thawed spermatozoa. In conclusion, supplementation of semen diluents with BHT can ameliorate preservability of goat sperm.     

Occurrence and Distribution of Potato Pests and Diseases in Kenya

Potato plays an important role in food security in Kenya but yields are low (<10 t/ha), and this is partly attributed to the lack of healthy planting material. This study is the first wide-scale survey to determine the occurrence and distribution of common potato pests and diseases in Kenyan seed (certified and quality declared) and ware crops. Potato crops growing on 101 farms in 21 districts were examined. Approximately 36% of plants in farmers’ fields sampled both during the long rains (main potato-growing season) and short rains seasons displayed virus-like disease symptoms. Six viruses (potato leafroll virus (PLRV), Potato virus A (PVA), potato virus M (PVM), potato virus S (PVS), potato virus X (PVX), and potato virus Y (PVY)) were detected using double antibody sandwich enzyme-linked immunosorbent assay in potato samples. Sequencing of polymerase chain reaction products from PVY-infected plants revealed the presence of recombinant strains of PVY (NTN and Wilga). Four aphid species, Macrosiphum euphorbiae, Aphis gossypii, Myzus persicae, and Aphis fabae, colonized potato in all districts, occurring in greater numbers west of the Great Rift Valley than to the east. There was a positive correlation between virus incidence and aphid numbers in the long rains (main) potato-growing season. PLRV, PVM, PVS, PVX, and PVY were detected in solanaceous weeds. Ralstonia solanacearum was detected in soils from 13 farms in 8 of the 18 districts surveyed. Approximately 38% of soil samples were infested with Meloidogyne spp. Phytophthora infestans isolates belonging to the US 1 and 2_A1 genotypes were identified. Although many economically important diseases are present in Kenya, the lower aphid incidence in districts east of the Great Rift Valley may indicate that these districts are more suitable for seed potato production.