首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
A modified “outside-in” roller bottle with a high ratio of surface area to volume was used to cultivate Giardia lamblia. Yields were high, more so when bottles were rotated at 6 rph (9.3 ± 4.0 × 108 trophozoites/bottle) than at 12 rph (4.2 ± 1.9 × 108 trophozoites/bottle). The method was more efficient than stationary tube culture with respect to utilization of culture medium; trophozoite concentration after roller bottle culture (1.7 ± 0.8 × 106 trophozoites/ml) was significantly higher (by a factor of 2.8) than concentrations obtained from stationary tube culture (0.6 ± 0.4 × 106 trophozoites/ml, P < 0.002). Increased yields from roller bottle culture were not accounted for by a reduction in mean trophozoite generation time (roller culture, 10.7 ± 1.2 hr; stationary tube culture, 10.3 ± 0.6 hr) but may be related to prolongation of the period of log phase growth or increased trophozoite survival. Trophozoite yields expressed per unit surface area were significantly higher from roller bottle culture (7.2 ± 3.1 × 105 trophozoites/cm2) than from stationary tubes (1.9 ± 1.0 × 105 trophozoites/cm2, P < 0.002). Attempts to cultivate G. lamblia in spin culture using polystyrene beads (Biosilon) as a microcarrier were unsuccessful, trophozoite growth being inhibited rather than promoted. Roller bottle culture of G. lamblia, however, is efficient, economical, and less laborious than stationary tube culture, particularly when more than 108 trophozoites are required.  相似文献   

2.
The association of giardiasis with the malabsorption of zinc remains controversial. This study investigated changes in serum zinc levels in Giardia-infected mice subjected to different dietary zinc regimens. Thirty-five mice (strain C3H/HeJ) were randomly categorized into two groups. The first group was inoculated with 5 × 106 Giardia trophozoites (n = 18), and the second group remained Giardia free (n = 17). Each group (Giardia infected and Giardia free) was randomly classified into three subgroups and given low (9 mg Zn/kg), normal (33 mg Zn/kg), and high levels (288 mg Zn/kg) of dietary zinc over a 2-week period for acclimation. Fourteen days post-Giardia infection, all of the mice were euthanized and blood samples were collected. The number of trophozoites was quantified (hematocytometer), and serum zinc levels were determined via atomic absorption spectrophotometry. Significant increases in the median weights were only found in the Giardia-free mice (p < 0.05). A higher final median weight was found in the Giardia-free group when compared with that of the Giardia-infected group given low dietary zinc (p = 0.013). In the Giardia-infected group with low dietary zinc, the geometric mean of trophozoites was 3,498 ± 101 (SE) per milliliter. The Giardia-infected group had lower serum zinc levels than did the Giardia-free group with the high dietary zinc regimens (p < 0.05). Our results are consistent with studies among human populations, but further studies are required to elucidate the actual mechanism governing the zinc–giardiasis interaction.  相似文献   

3.
The relationship between scale and body growth for emigrating Atlantic salmon, Salmo salar, smolts was previously not understood and therefore was examined in this study using mark-recapture techniques. The size of smolts at time of recapture was significantly greater than when marked (P = 0.0002). The growth in length of smolts emigrating 5 km over an average of 20 days was 7.7 ± 6.1 mm per day. Instantaneous somatic growth (G body) ranged from 7.0 × 10−4 to 5.1 × 10−3 (mean = 2.7 × 10−3 ± 1.3 × 10−3). The mean number of plus growth circuli present per scale was significantly greater for smolts when recaptured compared to when marked (P = 0.0014). The instantaneous growth rate of scales (G scale) ranged from 1.4 × 10−3 to 11.5 × 10−3 (mean = 6.6 × 10−3 ± 4.3 × 10−3). The relationship between body size and scale radius showed positive allometry rather than isometry. The relationship of G scale with G body showed positive allometry indicating that scales grew at a slightly faster rate than the body during the emigratory period.  相似文献   

4.
ABSTRACT. Giardia trophozoites and cysts, isolated from mammalian and avian hosts, were examined by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and by fluorescent light microscopy for the presence of microbial symbionts. Mycoplasma-like organisms were observed on the surfaces of trophozoites isolated from the prairie vole, laboratory rat, and beaver. Intracellular bacteria were observed by TEM in the trophozoites and cysts of G. microti and by fluorescence microscopy in trophozoites and cysts of Giardia spp. isolated from beaver, muskrat, great-blue heron, and the green heron. Trophozoites of G. muris from rat small intestine contained viral-like particles measuring 60 nm in diameter. These observations suggest that biological associations between Giardia spp. and diverse microbes may be more common than formerly appreciated. It also raises the possibility of transmission of these apparent symbionts, via the Giardia cyst, to other mammalian hosts including man.  相似文献   

5.
SYNOPSIS. Culture forms of Trypanosoma lewisi grown at 27 C in a diphasic blood agar medium resemble in structure the stage found in the invertebrate host. Cultures inoculated with approximately 1 × 106 trypanosomes/ml attain maximum populations of 2–7 × 107 organisms/ml after 5–6 days of incubation. The stationary phase persists 6–15 days. The decline of the population is of relatively long duration with approximately 1 × 106 viable organisms/ml present after 90 days. Variations in growth were attributed to the preparation of defibrinated heated rabbit blood incorporated into the culture medium. With inocula of 3.0 × 105 trypanosomes/ml there was a lag in growth not observed with larger inocula. Trypanosomes incubated at elevated temperatures had altered growth curves compared to organisms at 27 C. Agitation of cultures did not affect the growth or stationary phases, but hastened the population decline. Heated and unheated 5% (v/v) normal rat serum incorporated in the liquid phase of the medium altered the growth of the organisms. Heated serum caused a decrease in the population and an extended lag phase. The effects on growth were more marked with unheated serum suggesting that both heat-stable and labile components affect growth. Antisera from rats injected with live culture forms included in the liquid phase inhibited, while antisera from rats 24 days after infection with the blood stream forms had no effect on the growth of the culture forms. Antisera from rabbits immunized with sonicates of culture forms also altered the growth of the organisms in culture. Rabbit antisera prepared by immunization with sonicates of dividing and non-dividing blood stream forms had no effect on the in vitro growth. Antisera from animals immunized with rat blood and culture medium were also without effect. The immunologic implications of the data are considered and discussed.  相似文献   

6.
Borrelia duttonii strain 406 K, a causative agent of relapsing fever, could not be cultivated in vitro in currently available media for borreliae. We have developed an in vitro cultivation system by using SflEp cell cultures. The average increases of the number of borreliae, when inoculated with 1.0 × 105 organisms per ml from infected mice, were 23-fold and 150-fold in the primary culture and the 3rd subculture, respectively. Even a single borrelia could propagate in this cultivation system. This system will be useful for immunological and physiological studies on uncultivable Borrelia strains.  相似文献   

7.
Jeong Jun Kim 《BioControl》2007,52(6):789-799
The activity of entomopathogens on insect pests has been investigated for many species but the influence of entomopathogenic fungi on factors other than mortality relating to population increase has not been frequently studied. The influence of Lecanicillium attenuatum CS625 (=Verticillium lecanii CS625) on development and reproduction of cotton aphid (Aphis gossypii) was investigated. A conidia suspension of the isolate was applied onto first instar nymphs. Increased spore concentration did not significantly affect each nymphal stage, total nymphal period, pre-reproductive period and the age of first larviposition. A significant dose effect on reduction of life span, reproductive period and fecundity was observed in 1st and 3rd instars after spore application. When conidia were applied to 1st instars, life span was significantly reduced to 10.8 and 8.4 days at 1 × 104 and 1 × 108 conidia/ml, respectively from 12.2 days in the control. During the life span, total fecundity was 41 ± 7.3, 26 ± 0.8 and 22 ± 5.7 nymphs per female at 1 × 104, 1 × 106 and 1 × 108 conidia/ml, respectively compared with 51 ± 2.0 nymphs per untreated female. Reproduction period was also significantly shortened with increasing spore concentration. Application of spores to 3rd instars showed a similar trend. However, daily fecundity of individual aphids was not affected by spore dose. It was concluded that the isolate of L. attenuatum is able to affect populations of cotton aphid by reducing life span and total fecundity as well as by killing the aphids directly.  相似文献   

8.
Aoyagi H 《Biotechnology letters》2006,28(20):1687-1694
An index [kv: average isolation rate of viable protoplast (number/ml min)] was established to evaluate the optimal conditions for protoplast isolation from cultured plant cells. The optimal conditions for protoplasts isolation from Nicotiana tabacum BY2 cultured cells could be determined on the basis of the kv [31.7 × 103 (number/ml min)]. The colony-forming efficiency of the protoplasts was about 46%. The optimal conditions for protoplasts isolation from Catharanthus roseus [kv = 38.1 × 103 (number/ml min)] and Wasabia japonica [kv = 14.2 × 103 (number/ml min)] cultured cells could also be determined. Furthermore, a method for rapid regenerating cell wall of protoplast in liquid culture using alginate gel containing locust bean gum was developed.  相似文献   

9.
10.
Biofuels derived from non-crop sources, such as microalgae, offer their own advantages and limitations. Despite high growth rates and lipid accumulation, microalgae cultivation still requires more energy than it produces. Furthermore, invading organisms can lower efficiency of algae production. Simple environmental changes might be able to increase algae productivity while minimizing undesired organisms like competitive algae or predatory algae grazers. Microalgae are susceptible to pH changes. In many production systems, pH is kept below 8 by CO2 addition. Here, we uncouple the effects of pH and CO2 input, by using chemical pH buffers and investigate how pH influences Nannochloropsis salina growth and lipid accumulation as well as invading organisms. We used a wide range of pH levels (5, 6, 7, 8, 9, and 10). N. salina showed highest growth rates at pH 8 and 9 (0.19?±?0.008 and 0.19?±?0.011, respectively; mean ± SD). Maximum cell densities in these treatments were reached around 21 days into the experiment (95.6?×?106?±?9?×?106 cells mL?1 for pH 8 and 92.8?×?106?±?24?×?106 cells mL?1 for pH 9). Lipid accumulation of unbuffered controls were 21.8?±?5.8 % fatty acid methyl esters content by mass, and we were unable to trigger additional significant lipid accumulation by manipulating pH levels at the beginning of stationary phase. Ciliates (grazing predators) occurred in significant higher densities at pH 6 (56.9?±?39.6?×?104 organisms mL?1) than higher pH treatments (0.1–6.8?×?104 organisms mL?1). Furthermore, the addition of buffers themselves seemed to negatively impact diatoms (algal competitors). They were more abundant in an unbuffered control (12.7?±?5.1?×?104 organisms mL?1) than any of the pH treatments (3.6–4.7?×?104 organisms mL?1). In general, pH values of 8 to 9 might be most conducive to increasing algae production and minimizing invading organisms. CO2 addition seems more valuable to algae as an inorganic carbon source and not as an essential mechanism to reduce pH.  相似文献   

11.
The growth rates of 3 species of phytoplankton were found to be dependent on the vitamin B12 concentrations in the media. In batch cultures, the vitamin B12 half-saturation constants and standard errors were 0.39 ± 0.042 μμg/ml for Thalassiosira pseudonana (clone 3H), 1.69 ± 0.24 μμg/ml for Isochrysis galbana, and 2.77 ± 1.65 μμg/ml for Monochrysis lutheri. A chemostat was used to grow T. pseudonana with vitamin B12 as the controlling factor. In the chemostat the yield and standard deviation, 102 ± 21 × 104 cells/μμg vitamin B12, was the same as in the batch culture, 126 ± 13 ± 104 cells/μμg. The chemostat half-saturation constant, 0.26 ± 0.068 μμg/ml vitamin B12, and maximum growth rate were in agreement with those obtained in batch cultures. Vitamin concentrations for maximum growth, rates were greater than those calculated necessary from yield data to give observed population densities similar to those in natural waters. In the sea the effect of vitamin B12 concentration on growth rates may be complicated by low concentrations of other nutrients or the presence of inhibitors.  相似文献   

12.
The plant hormone, gibberellic acid (GA), stimulated growth of a marine diatom, Cyclotella cryptica Reimann, Lewin and Guillard. Four concentrations of GA (5 × 20 × 25 × and 35 × 10?6 g/mL) were added to axenic cultures of C. cryptica. Changes in cell densities, measured by cell counts and turbidimetric readings, confirmed that GA at 20 × 10?6 g/mL produced maximum stimulation. There was an increase in the total number of cells produced and a shorter lag phase of growth at this concentration. Coulter counter measurements of cell size, as well as ocular micrometer measurements, indicated there was no significant variation in cell volumes of GA grown cells over that of the controls.  相似文献   

13.
SYNOPSIS. The growth and reproduction of Allogromia laticollaris was studied. More schizozoites were generally produced in mixtures of food organisms than on single algal foods. In the presence of moderate numbers of bacteria, cultures with Phaeodactylum tricornutum, Chlorococcum sp., Nannochloris sp., and an unidentified chlorophyte (BL-1), added singly, were also highly productive. Schizogony was the dominant asexual form of reproduction. Binary fission and cytotomy also occurred in bacterized otherwise unfed controls. 35S and 32P are convenient labels for measuring growth of A. laticollaris when introduced into the system in the range of 1 × 104 - 1 × 105 dpm/ml (32P specific activity ~ 2.03 MCi/g; 35S specific activity ~ 95 μCi/g). Small allogromiids grew faster than did larger ones. By means of the Taylor series modification of the classical least-squares method, a continuous life-cycle representation was calculated for A. laticollaris for the conditions of the experiment. Four points of cell volume growth were maxima for reproduction: 1.0 × 107μ per organism for curve I; 2.2 × 107μ3 and 1.2 × 107μ3 for curve II; and 6.7 × 107μ3 for curve III.  相似文献   

14.
 This report describes a protocol for the regeneration of plants from protoplasts isolated from proembryogenic masses (PEMs) in a suspension culture derived from the nucellar callus of mango (Mangifera indica L. cv 'Amrapali'). The maximum yield (24.6±1.1×106), with 81.04±4.1% viable protoplasts per gram PEMs, was obtained with an enzyme mixture containing 1.2% cellulase, 1.0% hemicellulase and 0.6% pectinase. An optimum density of 5×104 cultured protoplasts per milliliter culture medium was required for the highest frequency (88.89±5.40%) of division. Dividing protoplasts developed into microcalli that proliferated on medium supplemented with growth regulators (auxins or kinetin alone, or auxins with kinetin) and produced somatic embryos after transfer to a growth regulator-free medium. The protocallus on 2,4-D-containing medium produced the maximum number (102.50±6.93) of somatic embryos. Maturation of somatic embryos depended upon the presence, and the nature and combination of growth regulators in the medium during proliferation of the callus. The mature somatic embryos germinated and developed into plants that were transferred to soil. Received: 1 April 1999 / Revision received: 27 July 1999 / Accepted: 23 August 1999  相似文献   

15.
Given the increase in the incidence of insulin resistance, obesity, and type 2 diabetes in children and adolescents, it would be of paramount importance to assess quantitative indices of insulin secretion and action during a physiological perturbation, such as a meal or an oral glucose‐tolerance test (OGTT). A minimal model method is proposed to measure quantitative indices of insulin secretion and action in adolescents from an oral test. A 7 h, 21‐sample OGTT was performed in 11 adolescents. The C‐peptide minimal model was identified on C‐peptide and glucose data to quantify indices of β‐cell function: static φs and dynamic φd responsivity to glucose from which total responsivity φ was also measured. The glucose minimal model was identified on glucose and insulin data to estimate insulin sensitivity, SI, which was compared to a reference measure, SIref, provided by a tracer method. Disposition indices, which adjust insulin secretion for insulin action, were then calculated. Indices of β‐cell function were φs = 51.35 ± 8.89 × 10?9min?1, φd = 1,392 ± 258 × 10?9, and φ = 82.09 ± 17.70 × 10?9min?1. Insulin sensitivity was SI = 14.19 ± 2.73 × 10?4, not significantly different from SIref = 14.96 ± 3.04 × 10?4 dl/kg·min per µU/ml, and well correlated: r = 0.98, P < 0.0001, thus indicating that SI can be accurately measured from an oral test. Disposition indices were DIs = 1,040 ± 201 × 10?14 dl/kg/min2 per pmol/l, DId = 33,178 ± 10,720 × 10?14 dl/kg/min per pmol/l, DI = 1,844 ± 522 × 10?14 dl/kg/min2 per pmol/l. Virtually the same minimal model assessment was obtained with a reduced 3 h, 9‐sample protocol. OGTT interpreted with C‐peptide and glucose minimal model has the potential to provide novel insight regarding the regulation of glucose metabolism in adolescents, and to evaluate the effect of obesity and interventions such as diet and exercise.  相似文献   

16.
The study purpose was the analysis of barbel Barbus barbus (L.) milt quality and quantity with regard to the time following stimulation with [(D‐Ala6, Pro9NEt)‐mGnRH+metoclopramide] i.e. Ovopel. Selected parameters such as total volume of milt (TVM, ml), volume of milt per kg of body weight (VOM, ml kg?1 b.w.), sperm concentration (×109 ml?1), total sperm production (TSP, ×109), osmolality (mOsm kg?1) and pH of seminal plasma were determined. Sperm motility was analyzed by the CASA system, i.e. the percentage of sperm motility (MOT, %) and their progressive motility (PRG, %), curvilinear velocity (VCL, μm s?1) and straight line velocity (VSL, μm s?1), amplitude of lateral head displacement (ALH, μm), and beat cross frequency (BCF, Hz). Milt was collected from 12 specimens (N = 12), with the first portion obtained 12 h following treatment with Ovopel (1 granule kg?1 b. w.). Subsequent portions of milt were taken at 24 h intervals, i.e. after 36, 60, 84, 108, and 132 h. The control group (Control, N = 12) was injected with 0.9% NaCl at 0.5 ml kg?1b.w. from which milt was taken 12 h post injection. The highest TVM, VOM and TSP values were recorded 12 h after Ovopel treatment (3.2 ± 0.7 ml, 36.7 ± 10.5 ml kg?1 b.w. and 39.1 ± 9.4 × 109, respectively); lowest values were recorded after 132 h (0.8 ± 0.4 ml, 11.1 ± 6.5 ml kg?1b.w. and 13.7 ± 7.5 × 109, respectively). The highest seminal plasma osmolality values (300.0 ± 42.6 mOsm kg?1) as well as the lowest sperm concentration (12.5 ± 1.5 × 109 ml?1) were observed 12 h after Ovopel treatment. No significant differences in the percentage of sperm motility (MOT) were noted during any of the periods after hormonal stimulation, however, a change in the character of their movement (PRG) was observed. The lack of significant differences (P > 0.05) in VCL and VSL values between 12 h and 60–132 h indicates that the lengthening of time does not lead to a decrease in sperm velocity and, therefore, is not likely to have a negative impact on their quality. The highest ALH (1.9 ± 0.2 μm) and BCF (11.5 ± 1.1 Hz) values were observed when the effect of stimulation was most noticeable, i.e. 12 h after Ovopel treatment. Based on the total milt volume and sperm production, the best time for milt collection from barbel is 12 h post‐hormonal treatment; 84 h post‐hormonal treatment, TVM, VOM, TSP and some CASA parameters decreased, which suggest the same aging process in sperm.  相似文献   

17.
Genomic DNA was extracted either directly from Giardia muris cysts seeded into environmental surface waters or from cysts isolated by immunomagnetic beads (IMB). A 0.171-kbp segment of the giardin gene was PCR-amplified following “direct extraction” of Giardia DNA from seeded Cahaba river water concentrate with moderate turbidity (780 JTU's), but DNA purified from seeded Colorado river water concentrates with high turbidity (2 × 105 JTUs) failed to amplify. However, if the cysts were first separated by the IMB approach from seeded Cahaba or Colorado river waters, and the DNA released by a freeze-boil Chelex?100 treatment, detection of G. muris by PCR amplification could be achieved at a sensitivity of 3 × 100 or 3 × 101 cysts/ml, respectively. If, however, the G. muris cysts used to seed even moderately turbid river waters (780 JTUs) were formalin treated (which is conventionally used for microscopic examination), neither direct extraction nor IMB purification methods yielded amplifiable DNA. Use of immunomagnetic beads to separate Giardia cysts from complex matrices of environmental surface waters followed by DNA release and PCR amplification of the target giardin gene improved the reliability of detection of this pathogen with the required sensitivity. Received: 23 April 1997 / Accepted: 4 August 1997  相似文献   

18.
Specific activity of aquatic bacteria, which indicates average heterotrophic activity per bacterial cell, was determined asV max per bacterium and turnover rate per bacterium for glucose mineralization at different sites (river and estuary) in north Humberside, northeast England.V max per bacterium ranged from 0.05×10−13 to 52.2×10−13 mg/h and turnover rate per bacterium from 0.05×10−8 to 88.3×10−8 ml/h. Highest mean values were found at river sites and the lowest at an outer estuary site, although there was considerable variation at each site and ranges from all sites overlapped. Also, ranges ofV max per bacterium from Humberside sites in general overlapped published ranges for sites in other geographical areas.V max per bacterium and turnover rate per bacterium were significantly correlated with some environmental variables, which suggests that they are of ecological significance.  相似文献   

19.
ABSTRACT. We studied galactose (Gal)-specific binding of the soluble purified 260-kDa Entamoeba histolytica adherence protein to glycosylation deficient Chinese hamster ovary (CHO) cell mutants. Our goal was to further define the lectin's functional activity and carbohydrate receptor specificity. The adherence protein was purified by acid elution from an immunoaffnity column; however, exposure of the surface membrane lectin on viable trophozoites to identical acid pH conditions had no effect on carbohydrate binding activity. Saturable Gal-specific binding of soluble lectin to parental CHO cells was demonstrated at 4°C by radioimmunoassay; the dissociation coefficient (Kd was 2.39 × 10?8 M?1 with 5.97 × 104 lectin receptors present per CHO cell. Gal-specific binding of lectin to Lec2 CHO cell mutants, which have increased N- and O-linked terminal Gal residues on cell surface carbohydrates, was increased due to an enhanced number of receptors (2.41 × 105/cell) rather than a significantly reduced dissociation constant (4.93 × 10?8 M?1). At 4°C, there was no measurable Gal-specific binding of the adherence protein to the Lec and IdlD.Lecl CHO mutants, which contain surface carbohydrates deficient in terminal Gal residues. Binding of lectin (20 μg/ml) to CHO cells was equivalent at 4°C and 37°C and unaltered by adding the microfilament inhibitor, Cytochalasin D (10 μg/ml). Gal-specific binding of the lectin at 4°C was calcium independent and reduced by 81% following adsorption of only 0.2% of the lectin to CHO cells. In summary, these findings indicate that the purified E. histolytica adherence lectin demonstrates saturable Gal-specific binding to 1–6 branched-N-linked and not O-linked galactose terminal cell surface carbohydrates; however, apparently only a small percentage of purified amebic lectin molecules actually possess galactose binding activity.  相似文献   

20.
Virioplankton and bacterioplankton abundance has been determined in the pelagic and littoral zones of the Rybinsk Reservoir during the ice-covered period. The role of viruses in heterotrophic bacterioplankton infection and mortality is assessed. At water temperatures between 0.3 and 0.9°C, the number of planktonic virus particles and planktonic bacteria varies from 37.1 × 106 to 84.1 × 106 particles/mL, (57.3 ± 2.1) × 106 particles/mL on average and from 2.50 × 106 to 6.11 × 106 cells/mL, (3.66 ± 0.16) × 106 cells/mL on average, respectively. The ratio of the virus number to the bacteria number varies from 8.8 to 27.9, being 16.5 ± 0.7 on average. Visually infected cells comprise 0.3–0.5% (1.5 ± 0.2% on average) of the total number of bacterioplankton. Infected bacterial cells contain from 5 to 107 (17 ± 4 on average) mature virus particles. The average virus-induced mortality of bacteria accounts for 13.0 ± 1.9% (variations range from 2 to 55%) of the daily bacterial production, indicating that viruses play an important role in the regulation of bacterioplankton production and abundance in the Rybinsk Reservoir during the ice-covered period.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号