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.     

Arbovirus isolations from mosquitoes in South Slovakia.

In the years 1973 and 1975 mosquitoes and some other Diptera (Tabanidae, Simuliidae, Hippoboscidae) were tested for virus. 13,924 mosquitoes, 75 horseflies and 60 blackflies were processed in 1973. Five strains of Tahyna virus were isolated from mosquito species Aedes vexans. 3,378 mosquitoes and 12 sheep keds were tested for virus in 1975. Twelve strains of Calovo virus were isolated from Anopheles maculipennis and one strain of Tahyna virus was obtained from Aedes vexans mosquitoes.     

Scanning electron microscopy of Setaria labiatopapillosa (Alessandrini, 1848) from cattle of Kazakhstan.

Surface structures of males and females of Setaria labiatopapillosa were studied by means of scanning electron microscopy at high magnification. It was found that the surface of buccal cavity, lateral appendages, vulva, anus and cloaca differs from the remaining cuticle. The shape of internal and external mouth papillae, amphids, deirids, postdeirid and phasmidial pore was studied. On the surface of the postdeirid are four small papillae arranged in a typical manner around the depression from which projects a needle-like formation. The ventral bands are formed by fine longitudinal parallel folds of the cuticle. Some anomalies in the localization of postcloacal papillae were observed.     

Nerine latent virus: some properties and serological detectability in Nerine bowdenii

Nerine latent virus (NeLV), first found inNerine bowdenii, may occur also in the otherNerine species investigated so far:N. sarniensis, N. flexuosa Alba, andN. Mansellii. Chenopodium amaranticolor, C. quinoa, andGomphrena globosa sometimes reacted with local lesions after mechanical inoculation with NeLV.Nicotiana clevelandii andHippeastrum were symptomless hosts. In this respect NeLV resembled the incompletely describedHippeastrum latent virus (HLV).Serologically NeLV was closely related to HLV and to carnation latent virus (CaLV), but differed from the latter in host plant reaction. A more distant relationship was observed with some other carlaviruses, wheareas NeLV also reacted with an antiserum to potato virus X.Depending on the lot, NeLV could be detected rather reliably with the micro-precipitin test inN. bowdenii Van Roon, but less well in 63. Better results were obtained with the microplate method of enzyme-linked immunosorbent assay (ELISA).The average particle length was 664 nm, the sedimentation coefficient 155 S and the buoyant density 1.298 g/cm³.NeLV can be considered as a member of the carlavirus group. On basis of priority HLV may be considered as NeLV.     

Phage typing of strains of Staphylococcus aureus from staphylococcal mastitis of dairy cows]

Of 62 strains of Staphylococcus aureus, isolated from milk samples of dairy-cows with staphylococcic mastitis from five agricultural enterprises, 68 p. c. could be typified as bacteriophages. In 42 typified strains, altogether 17 different phage pictures were found, in which case, in the different agricultural enterprises, the number of phage picture fluctuated between 1 and 8. Phage picture 42 D was found and dominated in all the agricultural enterprises. Of the typified strains, 76% could be ranked only into one phage group and 24% in the "M" group (= miscellaneous). Of 32 typified strains, 56.3% belonged to phage group IV, 25% to group III, 12.5% to group I, and 6.2% to group II.     

Phage typization of Staphylococcus aureus strains from staphylococcic mastitis of dairy cows]

Of 62 strains of Staphylococcus aureus, isolated from milk samples of dairy-cows with staphylococcic mastitis from five agricultural enterprises, 68 p. c. could be typified as bacteriophages. In 42 typified strains, altogether 17 different phage pictures were found, in which case, in the different agricultural enterprises, the number of phage picture fluctuated between 1 and 8. Phage picture 42 D was found and dominated in all the agricultural enterprises. Of the typified strains, 76% could be ranked only into one phage group and 24% in the "M" group (= miscellaneous). Of 32 typified strains, 56.3% belonged to phage group IV, 25% to group III, 12.5% to group I, and 6.2% to group II.     

Distribution of Cryptococcus neoformans in a pigeon habitat.

Samples of pigeon droppings were taken from 7 sites in a church tower contaminated with C. neoformans, and the distribution patterns of the fungus were studied. From various sites, 0 to 3 x 10(5) viable C. neoformans particles were recovered per one gram of dry excreta. The factors causing the different density of C. neoformans population in the habitat were: uric acid share of the total nitrogen, creatinine content, sunlight and probably pH. Chemical composition of the substrate is the primary factor in the distribution of C. neoformans in droppings.     

Age-related resistance to Pseudomonas syringae pv. tomato is associated with the transition to flowering in Arabidopsis and is effective against Peronospora parasitica

As plants mature it has been observed that some become more resistant to normally virulent pathogens. The ability to manifest the Age-Related Resistance (ARR) response in Arabidopsis to Pseudomonas syringae pathovars tomato (Pst) coincided with the transition to flowering in plants both delayed and accelerated in the transition to flowering. ARR was also associated with a change in PR-1 gene expression, such that young plants expressed PR-1 abundantly at 3 days post inoculation (dpi) while mature plants expressed much less. The Arabidopsis ARR response requires SA accumulation via isochorismate synthase (ICS1) [24]. ICS1 was expressed one dpi with virulent and avirulent Pst in both young and mature plants. The ARR response was also effective versus avirulent Pst providing an additional 4-fold limitation in bacterial growth. Arabidopsis ARR was found to be ineffective against two necrotrophs, Erwinia carotovora subspecies carotovora (bacterium) and Botrytis cinerea (fungus) and one obligate biotroph, Erysiphe cichoracearum (fungus). However, mature wild type, SA-deficient sid2 and NahG plants supported little growth of the obligate biotrophic oomycete, Peronospora parasitica. Therefore ARR to P. parasitica appears to be SA-independent, however the level of ARR resistance was somewhat reduced in these mutants in some experiments. Thus, there may be numerous defence pathways that contribute to adult plant resistance in Arabidopsis.     

Estimation of daily and total lactation milk yield of Chios ewes from single morning or evening records

Linear models were developed and evaluated for the estimation of daily (24 h) and total lactation milk yield of dairy sheep from single morning (am) or evening (pm) milking records. A total of 22,908 individual test-day milk records of 5110 ewes of the Chios breed, raised in 46 flocks, were used. Depending on the model, different daily milk yield estimates were derived for each ewe, accounting for lactation number, stage of lactation, season of previous lambing and interval between successive milkings. Daily milk yield was also estimated from doubling the single am or pm record. Actual and estimated yields were compared using root mean square errors (RMSEs), mean absolute differences, an accuracy parameter defined as the ratio of the actual yield variance over the sum of the variances of actual yield and absolute difference, and the product moment correlation between estimated and actual yield. Results were validated on independent datasets. Linear models resulted in less biased and more accurate estimates of the daily milk yield than simply doubling the am or pm record. Root mean square errors decreased by 7-37% and the mean absolute difference was reduced by 1-4% of the average daily yield. Higher accuracy and correlation were obtained from linear model application than doubling. Total lactation milk yield was predicted based on actual daily yield and compared to predictions based on estimated daily yield from am or pm records, as well as directly on single milking records. Root mean square errors obtained when daily yield had been estimated with linear models were 26-35% lower compared to doubling the am or pm yield and 0-13% lower compared to estimating the total lactation yield directly from single milking records. Linear model application also resulted in lower mean absolute difference and higher accuracy and correlation than doubling the am or pm record. Recording the yield of a single milking (am or pm) instead of both can benefit milk recording by reducing its cost and increasing farmer participation. In this context, linear models developed in the present study can be used for the accurate estimation of daily (24 h) and total lactation milk yield from single milking records.     

GAL‐ and VIP‐Immunoreactive Nerve Structures in the Ileum and Large Intestine of Pigs with Dysentery

The morphology, neurochemistry and function of intramural nerve structures in the mammalian gastrointestinal tract are relatively well known, but in normal, healthy individuals. The present study was aimed at investigating the chemical coding of nerve structures in the wall of the ileum and large intestine in normal pigs (n = 3) and in pigs undergoing dysentery (n = 6). Dysentery was evoked by artificial infection of the clinically healthy animals per os with Brachyspira hyodysenteriae. All the animals were deeply anaesthetized and transcardially perfused with 4% paraformaldehyde. The cryostat sections of the intestines were processed for double‐labelling immunohistochemistry using antisera against PGP 9.5, GAL and VIP. In the intramural plexuses of the control pigs, the percentage of GAL‐immunoreactive (GAL‐IR) perykarya varied from 11% (descending colon) to 19% (centrifugal turns of the ascending colon) whereas in the dysenteric pigs, it was distinctly higher, reaching from 28% (ileum) up to 48% (cecum). In the control animals, the percentage of VIP‐IR neuronal somata varied from 3% (descending colon) to 19% (ileum). In dysenteric pigs, it was from 6% (descending colon) up to 28% (cecum). In the muscular coat (MC) and mucous membrane (MM) of the normal intestine, very numerous GAL‐ and VIP‐IR nerve fibres were observed. The nerve fibres in the myenteric plexus (MP) were even more numerous than those in the muscular coat while in the outer (OSP) and inner (ISP) submucous plexuses, they were less abundant. In the dysenteric pigs, the nerve fibres found in MC, MP and OSP were less numerous, whereas those observed in ISP and MM were more abundant than those in the control animals. The present results suggest that GAL and VIP are involved in the regulation of inflammatory processes developing in the porcine gastrointestinal tract during dysentery.