Chromosomal evolution in salmonids: a comparison of Atlantic salmon, brown trout, and rainbow trout R-band chromosomes

Replication banding technique was applied to the chromosomes of Salmo salar, Salmo trutta, and Oncorhynchus mykiss. The in vitro technique has proved more advantageous than the in vivo technique due to a higher number of bands obtained. The comparison of these replication banding patterns has revealed that some chromosomes of Salmo trutta karyotype appeared associated with Salmo salar and Oncorhynchus mykiss karyotypes by single chromosomal rearrangements.     

临床新技术评价指标体系的建立及应用研究

目的 建立科学的临床新技术评价指标体系,为临床新技术管理提供依据。方法 结合管理实践,通过文献及政策调研、专家咨询、实践验证等方法进行研究。结果 建立了临床新技术定性与定量相结合的评价指标体系,采取同行函审和集中评定相结合的办法进行了实证研究,探索了临床新技术管理的有效方法。结论 评价体系的建立和应用为临床新技术的科学管理提供了依据,推动医疗技术的进步和医院的发展。     

Observations on paraffin baiting as a laboratory diagnostic procedure in nocardiosis

The efficacy of paraffin bait technique in the isolation ofNocardia asteroides from clinical specimens has been investigated. In a comparative study 1091 clinical specimens, mostly sputa and bronchial aspirates collected from 639 patients of bronchopulmonary diseases and 11 of meningitis, were examined by paraffin baiting and the conventional technique. Thirty-six clinical specimens originating from 12 of the patients yieldedN. asteroides by the paraffin bait technique but only 4 by the conventional technique. Approximately 95 % of 125 sputum samples inoculated withN. asteroides yielded the pathogen by paraffin baiting as against 49 % by the conventional technique. Paraffin baiting was more productive than the conventional technique in the isolation ofN. asteroides from mixed suspensions with a number of fungi and bacteria. It is concluded that paraffin baiting can be profitably adopted as a suitable technique for the isolation ofN. asteroides from clinical specimens, such as, sputum, gastric lavage, etc., which are often contaminated. The technique has no particular advantage with non-contaminated specimens.This work forms a part of the Ph. D. thesis of S.K.M. submitted in 1971 to the University of Delhi, and was presented at the Vth meeting of ISHAM held in Paris from 5th–10th July, 1971.     

An appraisal of seed enumeration and videographic techniques for determining seed removal rates by birds

We examined the efficacy of seed enumeration and videographic techniques for determining seed removal by birds from indigenous (Chrysanthemoides monilifera and Olea europaea subsp africana) and alien (Lantana camara and Solanum mauritianum) shrubs at different study sites in the Cape Floristic Region. The seed enumeration technique involved counting the numbers of fruits and associated seeds removed monthly by birds, excluding those naturally abscised, from the shrub canopy. The videographic technique involved visual counts from images of the numbers of fruits and associated seeds consumed by birds over specific time intervals captured by a digital camcorder. Daily seed removal rates by all birds, irrespective of species, measured by both techniques were similar with no significant interactions evident between measuring techniques, site and shrub species. Both techniques displayed higher seed removal from tiny‐seeded S. mauritianum than other shrub species; this was also evident among individual bird species. However, the seed enumeration technique was unable to discriminate between foraging organisms, contamination of traps by wind‐blown fruits abscised from neighbouring branches and fruit theft from the canopy and the traps. In contrast, the videographic technique provided permanent visual and time‐lapse records for individual foraging bird species allowing greater measurement precision and interpretation of fruit removal behaviour by birds. We recommend use of the videographic technique over the seed enumeration technique for studying vertebrates’ seed removal in a detailed manner.     

Development of an in vitro incubation technique for the estimation of the utilizable crude protein (uCP) in feeds for cattle

An in vitro incubation technique based on the first stage of the in vitro digestion technique published by Tilley and Terry (1963) was developed to estimate the utilizable crude protein (uCP) of single feeds and feed mixtures as non ammonia‐N after 24 h of incubation. The results of 25 feed samples showed that there was a significant relationship between the uCP values calculated by regression based on in vivo data sets (Y, CP [g‐kg^‐1 DM]) and those measured by the in vitro incubation technique (X, CP [g‐kg^‐1 DM], 24 h incubation):

y=0.85x+18.0, r²=0.84, P≤0.001.

It was concluded that it can be possible to determine the uCP value of single feeds or feed mixtures by this in vitro incubation technique and to estimate the uCP value of feeds by this regression equation.     

Control of Liriomyza langei on chrysanthemum by Diglyphus isaea produced with a standard or modified parasitoid rearing technique

Abstract: Overproduction of male parasitoids during mass rearing will increase costs for biological control because wasp shipments contain fewer females and only females kill hosts directly. We have developed a rearing technique capable of significantly reducing male‐biased sex ratios in Diglyphus isaea (Walker) (Hym., Eulophidae), a commercially reared parasitoid of agromyzid leafminers. In this study, we examined the effect of rearing technique on the efficacy of D. isaea for biological control of Liriomyza langei Frick (Dip., Agromyzidae) on chrysanthemum, Dendranthema grandiflora Tzvelev var. ‘Miramar’. We produced D. isaea on mixtures of small and large hosts (our modified technique) or on only large hosts (simulating commercial mass‐rearing) and compared: (1) control of L. langei with D. isaea produced by the two rearing techniques, and (2) damage and yield of unprotected and protected plants. The two rearing techniques produced similar numbers of waSPS per rearing cohort, but the ‘modified’ technique reduced the proportion of males by approximately 13%. The two techniques also produced females of similar size, but the ‘modified’ technique produced smaller males. In greenhouse trials simulating leafminer infestations of potted chrysanthemums during commercial production, we found no significant differences between the levels of control obtained by releasing identical numbers and sex ratios of adult waSPS produced by either rearing technique. Mine counts on plants protected by waSPS of either rearing history were similar and around 30–70% less than unprotected plants during most of the 11‐week crop cycle. At crop harvest, more than half of the foliage on protected plants was undamaged compared with <10% on unprotected crops. Damage to the flower stems of protected plants was relatively light in the top half of the canopy compared with the bottom half. Protected plants were around 10–15% taller and produced twice as many flower buds compared with unprotected plants. Our ‘modified’ rearing technique can reduce overproduction of males in D. isaea with no compromise in biological control efficacy. Adoption of our rearing technique by commercial insectaries could reduce implementation costs for not only D. isaea but also other parasitoids that show host‐size‐dependent sex allocation.     

PHOTOGRAPHIC VISUALIZATION OF FLORAL COLORS AS PERCEIVED BY HONEYBEE POLLINATORS

An inexpensive photographic technique for visualizing the ultraviolet and visible wavelengths perceived by honeybees is described. Using a standard daylight-balanced color slide film and illumination from blacklight and filtered daylight fluorescent lamps, a recording balance was achieved which approximates the spectral sensitivity of the honeybee eye. The technique was used to illustrate floral features among Rudbeckia species and among color morphs of Phlox. The Rudbeckia have inflorescences that are similar in visible coloration but distinctive in ultraviolet patterning and Phlox color morphs are distinctive in visible coloration but similar in ultraviolet patterning. The efficacy of the technique was judged from comparison with in vivo reflectance spectra of the floral subjects. Generally, photographic visualizations of entomophilous flowering plants portray only the ultraviolet or the visible components of floral coloration. This technique emphasizes the importance of considering the entire spectrum of floral colors relevant to most insect pollinators.     

Extra thin alginate film: an efficient technique for protoplast culture

Summary. This paper reports an efficient protoplast culture technique, the “extra thin alginate film” technique. The development of this improved method of protoplast culture was an outcome of an assessment of the efficiency and shortcomings of various protoplast culture techniques. The efficiency of this technique was evaluated with two model plant systems, viz., Nicotiana tabacum and Lotus corniculatus, and a comparison was made with the “thin alginate layer” technique, another efficient protoplast culture system. Results indicate that the culture technique with extra thin alginate film is as efficient as the technique with thin alginate layer, with many additional advantages. The present innovation overcomes most of the limitations of protoplast culture techniques described so far and can now be applied to a wide variety of crops to check its general applicability. Correspondence and reprints: Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar 143 005, India.     

A technique for quantitative evaluation of ectoparasitic mites and insects of domestic animals

A modified KOH dissolution technique using Tween 80 has been developed for the diagnosis and quantitative evaluation of ectoparasitic mites and insects of veterinary importance. Fifty intact skin segments from ten different domestic animals were artificially inoculated in vitro with ten specimens each of the mitesSarcoptes scabiei, Chorioptes bovis, Psoroptes ovis andDemodex caprae and insectsHaematopinus eurysternus andBovicola bovis. Sixty-seven to 88% of the mites and 80–99% of the insects were recovered by this technique. Using this method, parasitic mites and insects were found and identified in all skin scrapings from ten naturally infested animals. Using Cook's dissolution technique in our experimental design, 63.4% ofC. bovis and 65% ofS. scabiei mites were recovered.     

Reaction in vitro of synthetic oxytocin and lysine-vasopressin with the pseudoisocyaninchloride technique used for the demonstration of neurohypophysial neurosecretory material

Summary The performic acid (or acid potassium permanganate) pseudoisocyaninchloride technique (PSICN), when applied to formol-fixed paraffin-embedded sections of the hypothalamo-neurohypophysial system, causes to fluoresce intensely material with the same distribution as chrome-alum haematoxyphil neurosecretory material.It has been claimed that in vitro this technique does not react with oxytocin, but does react with neurophysin, the carrier (or precursor) polypeptide. Now however, using Rodeck's agar technique, which mimics closely conditions applying in tissue-sections, we have demonstrated that the PSICN technique will react in vitro with the octapeptide neuro-hormones, oxytocin and vasopressin.     

An improved method for mass rearing Paratheresia claripalpis for biological control of lepidopteran stalk borers

An improved method for mass rearing Paratheresia claripalpis Van der Wulp (Diptera: Tachinidae), for biological control of lepidopteran stalk borers is described and compared to the commonly used method. The improved method was found to be superior by being less labour intensive and by producing 2.25 times more parasites from the same number of hosts.

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Amélioration de la technique d'élevage de masse de Paratheresia claripalpis pour la lutte biologique contre Diatraea saccharalis

Résumé Une technique d'élevage de massa de la tachinaire, P. claripalpis Van der Wulp, est décrite et comparée à la technique classique. Cette technique correspond à une amélioration puisqu'elle demande moins de travail et produit 2.25 fois plus de parasites pour le même nombre d'hôtes.

     

Detection and identification ofListeria monocytogenes from milk and cheese by a single-step PCR

Primers ofiap gene were used as a target to develop a PCR technique for detectingListeria monocytogenes in milk and cheese. The PCR technique gives good results in the detection ofListeria monocytogenes either in artificially or naturally contaminated foodstuffs and has a high sensitivity and specificity. Application of this rapid diagnostic tool could provide further information about the spread ofL. monocytogenes in milk and cheese.     

A new chromosome fluorescence banding technique combining DAPI staining with image analysis in plants

In this study, a new chromosome fluorescence banding technique was developed in plants. The technique combined 4,6-diamidino-2-phenylindole (DAPI) staining with software analysis including three-dimensional imaging after deconvolution. Clear multiple and adjacent DAPI bands like G-bands were obtained by this technique in the tested species including Hordeum vulgare L., Oryza officinalis, Wall & Watt, Triticum aestivum L., Lilium brownii, Brown, and Vicia faba L. During mitotic metaphase, the numbers of bands for the haploid genomes of these species were about 185, 141, 309, 456 and 194, respectively. Reproducibility analysis demonstrated that banding patterns within a species were stable at the same mitotic stage and they could be used for identifying specific chromosomes and chromosome regions. The band number fluctuated: the earlier the mitotic stage, the greater the number of bands. The technique enables genes to be mapped onto specific band regions of the chromosomes by only one fluorescence in situ hybridisation (FISH) step with no chemical banding treatments. In this study, the 45S and 5S rDNAs of some tested species were located on specific band regions of specific chromosomes and they were all positioned at the interbands with the new technique. Because no chemical banding treatment was used, the banding patterns displayed by the technique should reflect the natural conformational features of chromatin. Thus it could be expected that this technique should be suitable for all eukaryotes and would have widespread utility in chromosomal structure analysis and physical mapping of genes.     

Development of a transplantation technique of Cystoseira amentacea var. stricta and Cystoseira compressa

On the North-Western Mediterranean coastline, the genus Cystoseira (Fucales) has a key role on upper infralittoral community structure. Cystoseira amentacea var. stricta and C. compressa are the most common superficial Cystoseira species. In this area, the coastline is subjected to a strong anthropogenic pressure which has caused severe Cystoseira population decreases. The present study consists in the development of a cheap and easy to use transplantation technique of C. amentacea var. stricta and C. compressa in order to either restore former natural populations or help their settlement on artificial substrates. The thalli were fixed with epoxy glue in holes made by a drill. The success of the technique was assessed by counting the number of remaining transplanted thalli after 3 weeks, 3 months and 6 months. Results were found encouraging with 75% of survival for both species after 6 months. Moreover fertile fronds were observed on the transplanted thalli showing the harmlessness of the technique.     

Histochemical lectin affinity technique by means of FITC-labeled serum protein fractions

Summary A two-step affinity technique is described for light microscopic demonstration of the Concanavalin A, Agaricus bisporus lectin and Ricinus communis lectin binding sites by means of various FITC-labeled human and rabbit serum protein fractions. Experiments for the visualization of the Lens culinaris lectin and the Pisum sativum lectin binding sites gaves negative results. The technique consist of two reaction steps which involve the incubation of tissue sections in the lectins followed by the visualization of receptor-bound lectins with FITC-labeled serum protein fractions basing on their carbohydrate content. The specificity of the technique could be demonstrated by the addition of the hapten or by incubation in the FITC-labeled serum protein fractions only. In contrast to the direct or indirect staining methods only very small amounts of purified lectins are necessary.     

A simple and non-radioactive technique to study the effect of monophosphoesters on matrix vesicle-mediated calcification

A simple and non-radioactive technique based on O-cresolpthalein complexone assay was developed to study in vitro non-radioactive calcium (⁴⁰Ca) deposition by isolated matrix vesicles. Using this technique, the effect of various phosphoester substrates including ATP, AMP and β-GP on in vitro MV-calcification was studied. O-cresolpthalein complexone assay with non-radioactive calcium demonstrated that AMP or β-GP were more effective in promoting calcium deposition by isolated MVs than ATP. The application of this nonradioactive technique, which is highly sensitive and simple, would offer a useful alternative approach to the routinely used radiometric biomineralization assay which employs radioactive ⁴⁵Ca. Published: December 16, 2004.     

Sequential scanning electron microscopy of a growing plant meristem

Summary A two-step replica technique has been developed for sequential study of the epidermal cell pattern of a living plant by scanning electron microscopy. This method is nondestructive, allows periodic high resolution observation of the same developing tissue, and can precede use of any destructive technique, such as transmission electron microscopy. The replicas can be trimmed allowing observation of occluded surfaces, such as the areas between leaves, which are inaccessible in continuousin vivo studies. Here we study the developing leaf primordium ofGraptopetalum and discuss potential uses of the technique.     

Evaluation of Ssp I polymerase chain reaction assay in the detection of Wuchereria bancrofti infection in field-collected Culex quinquefasciatus and its application in the transmission studies of lymphatic filariasis

Abstract: Ssp I polymerase chain reaction (PCR) assay, developed for Wuchereria bancrofti, was evaluated for its sensitivity in detecting infection in the vector, Culex quinquefasciatus, in the field. The evaluation of the assay was carried out using pools of vector mosquitoes collected from areas under filariasis survey and control trial projects, in comparison with the standard dissection and microscopy technique. In the filariasis survey area the infection rate as determined by the dissection and microscopy technique was 0.89% whereas it was 1.7% by PCR assay. In the Bacillus sphaericus trial area the infection rates as assessed by the conventional technique were 6.6 and 4.5% in the treated and check areas, respectively, whereas those obtained by the PCR assay were 4.7 to 2.2%. Although the infection rates determined by the PCR assay are slightly higher or lower than the rates obtained by the conventional technique, the difference was not statistically significant (P=0.451 for filariasis survey area, and P=0.203 and 0.161 for B. sphaericus trial area). When the pool size of Cx. quinquefasciatus was increased to 50 the sensitivity of the PCR was affected. The changes in infection rates as influenced by the antifilarial chemotherapy were similar when determined by PCR assay and the standard method. The major advantage of the PCR assay over the conventional technique is that thousands of mosquitoes can be processed within a short duration and this attribute has potential application in rapid assessment of disease prevalence and monitoring of the transmission dynamics.     

Restoring Quercus pyrenaica forests using pioneer shrubs as nurse plants

Question: How to improve reforestation success of Quercus pyrenaica. Location: 1800 m a.s.L, southern Spain. Methods: One‐year‐old Quercus pyrenaica seedlings were planted using two treatments: (1) bare soil, using a 30‐cm diameter augur bit (conventional technique) and (2) under the canopy of a pioneer shrub, Salvia lavandulifolia, using a 12‐cm diameter augur bit. Survival and growth were monitored for six years. Our hypothesis is that the use of shrubs as nurse plants is an alternative technique of reforestation with higher success than traditional techniques, in which pre‐existing vegetation is usually considered a source of competition. The rationale for the study was that for environments with a dry season, pre‐existing vegetation buffers summer drought stress, ameliorates the water status of seedlings and thus usually increases seedling recruitment. Results: Quercus survival was 6.3 × higher when planted under individuals of the pioneer shrub as compared to open areas. Quercus seedlings under shrubs also had shoots 1.8 X longer, while the number of shoots per plant did not differ among treatments. The first summer was the period with the highest mortality (49.1% of seedlings). Summer drought was the main cause of mortality. Conclusions: The use of shrubs as nurse plants for Q.pyrenaica reforestation is a viable technique to increase establishment success. The technique could be similarly useful in other environments with a dry period and for other Quercus species. In addition, this technique offers the advantage of following natural succession, thus minimizing the impact in the community.     

Some aspects of microbiological stimulation in a soil treated with streptomycin

Summary An investigation was made to follow the effect of streptomycin on the soil microflora using a direct method of observation together with plate counting technique on some selective media.It was reported that the bactericidal effect of streptomycin in the first week of application to soil, has masked an early stimulatory effect manifested by stimulation of two bacterial species appearing on mannitol nitrogen-free agar plates. One of these bacterial species identified as Pseudomonas sp. was able to fix small but significant amounts of nitrogen. The direct observation technique enabled to observe some aspects of microbial stimulation in soil. Stimulation of large cocci, identified as Hansenula sp., long bacterial rods, and Fusarium solani were successfully observed by such technique. Enrichment technique helped in isolating Hansenula sp. and F. solani; F. solani formed a mat on the surface of treated soil 50 days after streptomycin application. The recorded aspects of stimulation were claimed to be due to microbial equilibrium disturbance as a result of applying streptomycin to soil. Suggestion for further investigations was offered to verify the possible significance of such aspects of stimulation in controlling soil-borne pathogens.