Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries.This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt.Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.     

Risk factors and indicators of reduced welfare of grazing dairy cows from selected smallholder dairy farms in Midlands Province,Zimbabwe

Zimbabwe’s smallholder dairying faces many challenges that affect the welfare of dairy cows; however, the status of this welfare has not yet been determined. A cross-sectional study was conducted in Midlands Province on 41 active smallholder dairy farms with 86 cows in winter and summer to determine the risk factors and indicators to impaired cow welfare. These objectives were achieved using farmer questionnaires and direct observations. Eleven percent of the observed cows were severely lame in summer while only 5% were severely lame in winter. Lameness was significantly associated with season, absence of shade, breed, and low dipping frequency; 58% of the cows had low body condition scores (BC?<?3) and this low BC was associated with low frequency of protein (p?=?0.002) and vitamin (p?=?0.012) supplementation recorded in more than 52% of the farms visited. In winter, only 11% of the observed cows were heavily soiled (score 3), while in summer 64% of the cows were heavily soiled and this was associated with slurry accumulation in more than 80% of the observed cattle pens as well as the study season (p?<?0.001). A quarter (26%) of the studied animals had visible teat lesions on the teat skin and this was associated with the type of lubricant used (p?=?0.011). Only 34% of the cows allowed an approaching stockman to touch them and this was associated with shouting (p?=?0.012) and whipping of cows (p?=?0.002). The study concluded that welfare of dairy cows was poor in most of the smallholder dairy farms studied.     

The establishment of Taraxacum officinale plants in grassland

Factors involved in the establishment of Taraxacum officinale (dandelion) seedlings in grasslands and plant mortality in the first growing season were studied during 2008–2011. The study was conducted at 11 grassland sites that differed in the quantity of seed rain and the intensity of seedling predation by slugs. In April, of each study year, 10 plots open to the sedimentation of naturally dispersed seeds were established at each site, five plots of which were also sown with 250 T. officinale seeds each. The amount of natural seed rain, slug feeding activity and number of plants in each plot were then recorded at monthly intervals. Seed rain and seedling predation accounted for c. 50% of the variation in seedling establishment. Populations of established seedlings then declined through October, at which point they were at 25% of the original abundance on average. The percentage of plants that survived varied among sites and years. The temporal and spatial variation in T. officinale establishment and survival of young plants was enormous, even within a small area. Predation of seedlings by slugs can effectively prevent the establishment of T. officinale in grassland. This only occurred, however, at slug abundances not acceptable under conditions of public access.     

Factors affecting seed dormancy and germination of Johnsongrass, Sorghum halepense (L.) Pers.

Seeds of Johnsongrass [Sorghum halepense (L.) Pers.] germinated to higher percentages (20–30% higher) when incubated at 28 and 35° C than at 10 or 22° C. After-ripening was accelerated by dry storage of these seeds at 50°C. Seeds pre-chilled at 6°C for 2–4 weeks followed by incubation at 28°C germinated 40–60%. Light effects on germination were related to incubation temperatures; inhibitory at 22°C; no response at 28°C; and stimulatory at 35°C. Effects of gibberellin A₃ (GA₃) also varied depending on incubation temperature, sodium hypochlorite (NaOCl) immersion and light conditions. Immersion of dry seeds in either 700 mM NaOCl, 900 mM H₂O₂ or concentrated H₂SO₄ before incubation in water was effective in breaking dormancy. This result suggests the modes of action of H₂SO₄ in the termination of dormancy may be similar to those of NaOCl and H₂SO₄ as previously suggested by Hsiao & Quick (1984), that is by modification or scarification of the hull or seed coat membranes, and also by the supply of additional oxygen to the seed.     

Outbreak of clinical listeriosis in sheep: evaluation from possible contamination routes from feed to raw produce and humans

We report the results of clinical and microbiological investigations on Listeria monocytogenes infections in a flock of 55 sheep and describe the implications for the safety of the raw milk and raw-milk cheeses produced in the on-farm dairy. The outbreak was caused by feeding grass silage, which was contaminated with 5 log10 CFU L. monocytogenes/g. Clinically, although having been fed from the same batch of silage, abortive (nine ewes), encephalitic (one ewe) and septicaemic (four ewes) forms of listeriosis were observed during the outbreak phase. As the starting point of feeding the contaminated silage was known we could calculate an incubation period of 18+/-2 and 26 days for the abortive and the encephalitic form of listeriosis, respectively. Pathologically, the septicaemic cases suffered from Listeria accumulation at comparable numbers in visceral organs but not in the brain. Only a single ewe developed central nervous symptoms and a rhomb-encephalitis was immunohistologically confirmed. In this case the infection proceeded from the nasal mucosa into the brain, with no infections of the liver, spleen and other visceral organs. Sampling of the cheese production chain, the farm environment and the persons living at the farm revealed the exposure of a farm-worker to an isolate genetically indistinguishable from the outbreak clone, obviously through the consumption of faecally contaminated bovine raw milk. The cheese under processing was free of Listeria because, as a result of intensive consultations, the farmer ensured a proper acidification of the cheese. The epidemiological findings suggest that food safety matters should be assessed in any case where infection of food-producing animals with potential human pathogens is observed.     

Effect of a 1800 MHz electromagnetic field emitted during embryogenesis on chick development and hatchability

The level of artificial electromagnetic field (EMF) has steadily increased with the development of human civilization. The developing chicken embryo has been considered a good model to study the effects of EMF on living organisms. The aim of the study was to determine the effect of a 1800 MHz electromagnetic field during embryogenesis on the frequency of chick embryo malformations, morphometric parameters of the heart and liver and concentration of corticosterone in blood plasma, lipid and glycogen content in the liver of newly hatched chicks. A 1800 MHz EMF was found to shorten the duration of embryogenesis (earlier pipping and hatching of chicks) while having no effect on the quantity and quality of chicks and on increasing the incidence of embryo malformations. Exposure of chick embryos to EMF caused decreases in relative heart weight and right ventricle wall thickness. The pipping and hatching of chicks can be accelerated by stressful impact of EMF, which is confirmed by a significant increase in plasma corticosterone concentrations and decrease in fat and glycogen in the liver of chicks exposed during embryogenesis on the electromagnetic field with a frequency of 1800 MHz.     

Immunohistochemical studies on the splenic lobe of the pancreas in young Japanese quails (Coturnix c. japonica)

The splenic lobe (Lobus splenicus) of the pancreas of young meat-type quails (Coturnix c. japonica) was examined by immunohistochemical and light microscopic methods. The endocrine cells are mainly grouped as alpha, beta and mixed islets. A large region consisting of alpha cells is located in the central region of the splenic lobe whereas numerous beta islets are detected in the periphery of the splenic lobe. Alpha islets are in the majority composed of toluidine blue positive A cells and a few toluidine blue negative D and / or avian pancreatic polypeptide (APP) endocrine cells. Beta islets contain only a few toluidine blue negative B and a few D cells. Immunohistochemical staining of the splenic lobe reveal in the centre of beta islets numerous insulin immunoreactive cells and scarcely in alpha islets, exocrine tissue and / or among acinar cells. Somatostatin immune-reactive cells form a circular layer in the periphery of beta islets whereas these cells are uniformly distributed throughout the alpha islet parenchyma and exocrine tissue. In conclusion, the morphology but also the endo- and exocrine functions of the splenic lobe of quails are similar to observations in other avian species such as chicken, duck, goose and pigeon.     

Rhabdovirus-like particles in the sporocyst of the trematode Brachylaimus fuscatus from terrestrial mollusc Ponsadenia duplocincta

Rhabdovirus-like particles containing nucleoprotein in a helix with eight turns was demonstrated by transmission electron microscopy in subtegumental cells and syncytial layer of tegument in Brachylaimus fuscatus sporocyst. This is the first case of virus infection in trematodes. The possibility of transfer of the virus to intra-molluscan parasitic generations of the trematode is discussed with respect to the peculiarities of the life cycles of the family Brachylaimidae.