A study of pre- and postganglionic fibres in the intestinal nerve (Remak's nerve) of the chicken

Summary The mean peak CV's of two electrophysiologically defined groups of fibres in the intestinal nerve of the chicken have been determined.One group of fibres is constituted by the processes of enteric cholinergic neurones which project along the side branches of the intestinal nerve and synapse within the nerve trunk. These preganglionic fibres have a mean peak CV (at 40 °C) of 0.31 m·s^–1.The other group is made up of fibres of postganglionic neurones which project orally along the nerve trunk. The results suggest that some postganglionic neurones project only as far as the next ganglion whilst others project beyond the next two ganglia for distances greater than 5 mm. The postganglionic fibres have a mean peak CV (at 40 °C) of 0.71 m·s^–1.These figures demonstrate that both pre- and postganglionic fibres are unmyelinated. The temperature coefficient (Q₁₀) for the CV of unmyelinated fibres in the intestinal nerve was 1.57.Abbreviations CAP compound action potential - CV conduction velocity - Q ₁₀ temperature coefficient     

The use of simultaneous sampling bottle and vertical net collections to describe the dynamics of a zooplankton population

Simultaneous collections of zooplankton were made at four stations in the Plover Clove Reservoir using Friedinger sampling bottles and vertical Nakai plankton net hauls. Comparison of the results obtained revealed certain obvious numerical and spatial anomalies, and it is suggested that these result from inherent characteristics of the two types of apparatus together with behavioural responses and physical attributes of the individual zooplankton species. Statistical correlation between these two methods was generally good in terms of the seasonal patterns of distribution of the population, but in terms of depth distribution and the relative abundance of the individual species of the population few significant correlations resulted. It is therefore suggested that unless merely seasonal trends are required, it is advisable to use more than one sampling apparatus to obtain valid data concerning the overall dynamics of such a zooplankton population.Department of Botany, University of Hong Kong     

Characterization and taxonomic placement of Rhizoctonia-like endophytes from orchid roots

Twenty-one Rhizoctonia-like fungal strains were isolated from the roots of four terrestrial orchid species from various locations in Hong Kong. The cultural morphology, nuclear number of the hyphal cell, pore ultrastructure, and RAPD and CAPS analyses of rDNA fragments revealed that most of these isolates were associated with the genera Ceratorhiza and Epulorhiza. RAPD analysis showed the presence of genetic diversity between the isolates from different hosts and locations. The compatibility between a selection of these Ceratorhiza and Epulorhiza isolates and 14 orchid species was determined using a symbiotic germination method. The germination and development of three orchid species, Arundina chinensis, Spathoglottis pubescens, and Spiranthes hongkongensis, were strongly stimulated by the Epulorhiza isolates. Habenaria dentata was found to form symbionts successfully with a Ceratorhiza isolate.     

Proteomic study of a model causative agent of harmful red tide,Prorocentrum triestinum I: Optimization of sample preparation methodologies for analyzing with two-dimensional electrophoresis

A comprehensive study to find the optimal sample preparation conditions for two-dimensional electrophoresis (2-DE) analysis of Prorocentrum triestinum, a model causative agent of harmful algal blooms (HABs) was carried out. The four major sample preparation steps for 2-DE: (a) cell disruption: i.e. sonication and homogenization with glass beads; (b) protein extraction : i.e. sequential and independent extraction procedures; (c) pre-electrophoretic treatment: these included (i) treatment with RNAase/DNAase or benzonase; (ii) ultracentrifugation to sediment large macromolecules such as DNA; (iii) desalting and concentration by ultrafiltration through a Microcon centrifugal filter device (MWCO: 3000 daltons); and (iv) desalting by a micro BioSpin chromatography column (MWCO: 6000 daltons); and (d) rehydration buffers, reducing agents and sample application in the first dimension isoelectric focussing were studied. Our results showed that sonication is easy to perform and resulted in a higher protein yield. Among the four extraction buffers, the urea containing buffers resulted in the extraction of the highest amount of protein while tris(hydroxymethyl)aminomethane buffers and trichloroacetic acid (TCA)/acetone precipitation allowed detection of a higher number of protein species (i.e. protein spots). Desalting by BioSpin and ultrafiltration have improved the 2-DE resolution of the water soluble fraction but have less effect on urea containing fractions. TCA/acetone precipitation was able to desalt all protein fractions independent of the extraction media, however extended exposure to this low pH medium has caused protein modification. Introduction of either DNase/RNase or benzonase treatment did not improve the discriminatory power of the 2-DE but this treatment did yield 2-DE with the clearest background. Proteolytic digestion was inhibited by addition of a protease inhibitor cocktail. Taken overall, a combination of sequential extraction and desalting by BioSpin chromatography for sample treatment before first dimension of 2-DE gave best results based on its simplicity and minimal protein loss. Finally, triscarboxyethylphosphine (TCEP) has performed well as a reducing agent in both the rehydration and equilibration buffers. The rehydration buffer found to be best in this study was 8.0 M urea, 2% 3-[(3-cholamidoprphyldimethylamino]-1-propanesulfonate, 4 mM TCEP and 1% immobilized pH gradient buffer. Subsequently, we applied this finding and performed 2-DE analysis on the soluble protein fractions extracted from light-starved cultured algal cells (nonblooming) and cultured cells grown under optimal conditions (blooming). 2-DE maps of these algal cultures were visibly different and many differentially expressed proteins were found.     

Female reproductive parameters in the Javan gibbon (Hylobates moloch)

Javan gibbons (Hylobates moloch) are one of the most endangered gibbon species in the world. Data on the reproductive biology of the species are almost nonexistent, and a general understanding of the female reproductive biology of this species is important for both ex situ and in situ conservation. Using 18 years of data from 11 captive individuals, we provide new information on the reproductive biology of Javan gibbons based on sexual swelling and menstrual bleeding, including reproductive development, interbirth intervals, and ovarian cycle lengths. Menarche and the onset of sexual swelling occurred at 6.2 and 6.5 years respectively, followed by a period of adolescent sterility of about 1.5 years. Average age at first birth was 8.8 years, and interbirth intervals were about 2.3 years, decreasing to 1.0 year during cases of infant mortality at or shortly after birth. Ovarian cyclicity was measured through periods between menstrual bleeding and sexual swelling. Menstrual bleeding indicates the start of a new ovarian cycle, while sexual swelling normally occurs near the time of ovulation. Menstrual bleeding intervals gave a cycle length of 25.6 days, while sexual swelling intervals gave a cycle length of 27.3 days. These both correspond closely to cycle lengths in other gibbon species, as well as hormonal studies in Javan gibbons. In particular, observing the presence/absence of swellings was found to be a useful and easy method to monitor female ovarian cycles, and could be a practical noninvasive technique for caretakers and researchers. Zoo Biol 29:449–456, 2010. © 2009 Wiley‐Liss, Inc.     

Enzymatic studies of the extracellular mucilage of two aquatic hyphomycetes,Lemonniera aquatica andMycocentrospora filiformis

The effect of three carbohydrate-digesting enzymes, β-glucuronidase, lyticase and α-mannosidase and three proteolytic enzymes, α-chymotrypsin, papain and pronase E, on the strength of conidial attachment ofLemonniera aquatica andMycoentrospora filiformis was determined using the LH_Fowler cell Adhesion Measurement Module. Carbohydrate-digesting enzyme treatments showed significant differences in number of attached and detached, conidia versus control samples; little or no effect was observed for the proteolytic enzymes. Scanning and transmission electron microscopy showed different degrees of mucilage digestion by the carbohydrate-digesting enzymes on the germ hyphae, hyphae subtending appressoria, and appressoria of the two species. The loss of mucilage integrity and decrease in mucilage thickness were more pronounced on the hyphal sheaths than on the appressorial sheaths. Lyticase caused the most severe damage to the mucilage and cytoplasm of both fungi, particularlyL. aquatica. β-Glucuronidase and α-mannosidase exhibited more effective mucilage digestion onM. filiformis than onL. aquatica. Results indicate that the mucilage of the two species is mainly polysaccharide, containing more β-1,3-glucans than β-glucuronide and α-mannosyl residues. Variability of mucilage composition exists between these species and also between different structures of the same fungus.     

球形棕囊藻的生长特性及生活史研究

对球形棕囊藻不同的藻株(香港株HK和汕头株ST)在实验室条件下分别进行了形态学,生活史及生长曲线的研究,结果表明:球形棕囊藻具有一个复杂的异型生活史,具有两种不同的生活形态:群体和游离的单细胞,香港株和汕头株二者单细胞呈球形或近球形,直径大多为3-9μm;群体呈中空的球形囊泡,细胞包埋在胶质囊中较均匀分布,但二者群体大小有较大差异。当培养达到一定阶段,群体衰亡破裂释放大量单细胞。批次培养中球形棕囊藻的生长周期约为20-30d,香港株最适生长温度接近25℃,最大比生长率为038;汕头株的最适生长温度接近30℃,最大比生长率为042,这说明温度是重要的生长限制因子之一。       

Role of fungi in freshwater ecosystems

There are more than 600 species of freshwater fungi with a greater number known from temperate, as compared to tropical, regions. Three main groups can be considered which include Ingoldian fungi, aquatic ascomycetes and non-Ingoldian hyphomycetes, chytrids and, oomycetes. The fungi occurring in lentic habitats mostly differ from those occurring in lotic habitats. Although there is no comprehensive work dealing with the biogeography of all groups of freshwater fungi, their distribution probably follows that of Ingoldian fungi, which are either cosmopolitan, restricted to pantemperate or pantropical regions, or in a few cases, have a restricted distribution. Freshwater fungi are thought to have evolved from terrestrial ancestors. Many species are clearly adapted to life in freshwater as their propagules have specialised aquatic dispersal abilities. Freshwater fungi are involved in the decay of wood and leafy material and also cause diseases of plants and animals. These areas are briefly reviewed. Gaps in our knowledge of freshwater fungi are discussed and areas in need of research are suggested.     

Mechanism of cytotoxic action of azaguanine and thioguanine in wild-type V79 cell lines and their relative efficiency in selection of structural gene mutants

The cytotoxic effects of azaguanine and thioguanine have been compared in two wild-type V79 cells. To achieve equitoxic effects in both cell lines a 10–20-fold higher concentration of azaguanine than thioguanine was required. Affinity of HGPRT for azaguanine was 10-fold lower than for hypoxanthine in both cell lines and was similar to that for thioguanine in V79S cells. Affinity for thioguanine differed by a factor of 3 in the two cell lines. The rate of cell kill by azaguanine was markedly slower than by thioguanine in both cell lines. Reduction of whole cell uptake of [¹⁴C]hypoxanthine incorporation by unlabelled azaguanine was only demonstrable after prolonged incubation periods as was incorporation of [¹⁴C]azaguanine into acid-insoluble material. Experiments with cell-free extracts indicated that hypoxanthine acts as a non-competitive inhibitor of the enzyme. The slow rate of dissociation of the HGPRT—azaguanine complex is reflected in the slow rate of killing of wild-type cells. Clones resistant to the cytotoxic effects of these analogues have been selected from both cell lines and have been shown to possess HGPRT with altered kinetic properties. Our data suggest that azaguanine and thioguanine may select for mutations at different sites on the HGPRT molecule in V79 cells and provide possible explanations for the differences in effectiveness of these two agents reported in other cell lines.