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1.
The present study was aimed at carrying out a calibration and a comparison of diagnostic accuracy of three faecal egg counts (FEC) techniques, simple flotation, McMaster and FLOTAC, in order to find the best flotation solution (FS) for Dicrocoelium dendriticum, Moniezia expansa and gastrointestinal (GI) strongyle eggs, and to evaluate the influence of faecal preservation methods combined with FS on egg counts. Simple flotation failed to give satisfactory results with any samples. Overall, FLOTAC resulted in similar or higher eggs per gram of faeces (EPG) and lower coefficient of variation (CV) than McMaster. The "gold standard" for D. dendriticum was obtained with FLOTAC when using FS7 (EPG=219, CV=3.9%) and FS8 (EPG=226, CV=5.2%) on fresh faeces. The "gold standard" for M. expansa was obtained with FLOTAC, using FS3 (EPG=122, CV=4.1%) on fresh faeces. The "gold standard" for GI strongyles was obtained with FLOTAC when using FS5 (EPG=320, CV=4%) and FS2 (EPG=298, CV=5%). As regard to faecal preservation methods, formalin 5% and 10% or freezing showed performance similar to fresh faeces for eggs of D. dendriticum and M. expansa. However, these methods of preservation were not as successful with GI strongyle eggs. Vacuum packing with storage at +4°C permitted storage of GI strongyle eggs for up to 21 days prior to counting. Where accurate egg counts are required in ovine samples the optimum method of counting is the use of FLOTAC. In addition, we suggest the use of two solutions that are easy and cheap to purchase and prepare, saturated sodium chloride (FS2) for nematoda and cestoda eggs and saturated zinc sulphate (FS7) for trematoda eggs and nematoda larvae.  相似文献   

2.
It has been over 100 years since the classical paper published by Gosset in 1907, under the pseudonym "Student", demonstrated that yeast cells suspended in a fluid and measured by a haemocytometer conformed to a Poisson process. Similarly parasite eggs in a faecal suspension also conform to a Poisson process. Despite this there are common misconceptions how to analyse or interpret observations from the McMaster or similar quantitative parasitic diagnostic techniques, widely used for evaluating parasite eggs in faeces. The McMaster technique can easily be shown from a theoretical perspective to give variable results that inevitably arise from the random distribution of parasite eggs in a well mixed faecal sample. The Poisson processes that lead to this variability are described and illustrative examples of the potentially large confidence intervals that can arise from observed faecal eggs counts that are calculated from the observations on a McMaster slide. Attempts to modify the McMaster technique, or indeed other quantitative techniques, to ensure uniform egg counts are doomed to failure and belie ignorance of Poisson processes. A simple method to immediately identify excess variation/poor sampling from replicate counts is provided.  相似文献   

3.
The present study was aimed to evaluate the influence of flotation solution, sample dilution, and the choice of McMaster slide area (volume) on the reliability of the McMaster technique in estimating the faecal egg counts of gastrointestinal (GI) strongyles and Dicrocoelium dendriticum in a composite sample of faeces from naturally infected sheep. Fourteen flotation solutions having densities between 1.200 and 1.450, and six sample dilutions, 1:10, 1:15, 1:20, 1:30, 1:40 and 1:50 were used. Each of the six dilutions was divided into 70 aliquots in order to have five replicates of each of the 14 flotation solutions at each of the six dilutions. For each McMaster slide, the GI strongyle and D. dendriticum egg counts were performed under one grid (McM 0.15 ml), two grids (McM 0.3 ml), one chamber (McM 0.5 ml), and both chambers (McM 1.0 ml). Mean eggs per gram (EPG) of faeces of GI strongyles and D. dendriticum were calculated and statistical analyses were performed on the resulting data. The type of flotation solution used significantly influenced the EPG in the GI strongyles and in the D. dendriticum egg counts. All the sucrose-based solutions at density between 1.200 and 1.350 floated more GI strongyle eggs than the others. With respect to D. dendriticum, only six solutions were capable of floating eggs and the potassium iodomercurate solution (density 1.440) floated more eggs than the others. The reliability of the McMaster technique regarding sample dilution was high for both GI strongyle and D. dendriticum EPG at 1:10 and 1:15, and then progressively decreased with increasing dilution. The reliability of the McMaster technique regarding the choice of the McMaster slide area (volume) was high for both GI strongyle and D. dendriticum EPG at the McMaster slide area (volume) of 1.0 ml, i.e. the total area of the McMaster slide. The EPG counts resulting from choosing any of the other three McMaster slide areas (volumes), i.e. McM 0.15 ml, McM 0.3 ml, or McM 0.5 ml, produced unreliable over-estimates. The findings of the present study show that the highest reliability of the McMaster technique for estimating GI strongyle and D. dendriticum egg counts in faeces from pastured sheep is obtained when using flotation solutions based on sucrose for GI strongyles, and potassium iodomercurate for D. dendriticum, dilutions which do not exceed 1:15, and the McMaster slide area (volume) of 1.0 ml.  相似文献   

4.
Faecal pellets from a sheep that was artificially infected with a monoculture of Haemonchus contortus were collected over a 2-h period in the morning. In the laboratory the faeces were thoroughly mixed by hand and 48 by 1 g aliquots of the pellets were sealed in plastic bags, from which the air had gently been expressed. The faecal worm egg count of the sheep was about 14,000 g(-1). Varying numbers of the bags were either processed for faecal worm egg counting (FEC) by the McMaster technique on day 0, or were stored at one of the following temperatures: about 4 degrees C, -10 degrees C or -170 degrees C before processing. The faecal aliquots that were frozen were thawed at room temperature after having been frozen for either 2 h or 7 days, and processing of aliquots maintained at 4 degrees C proceeded shortly after the samples had been removed from the refrigerator. A dramatic reduction in egg numbers was found in all the aliquots that were frozen at -170 degrees C before faecal worm egg counts were done, as well as in those frozen for 7 days at about -10 degrees C. Numerous empty, or partially empty, egg shells were observed when performing the counts in faeces that had been frozen. In contrast, there was no significant reduction in the numbers of eggs in aliquots maintained for 7 days in a refrigerator at +/- 4 degrees C before examination, when compared with others examined shortly after collection of the faeces. Since H. contortus eggs in faeces are damaged by freezing, some methods that can be used for short term preservation are outlined. It is concluded that all nematode egg counts from cryopreserved faeces (whether in a freezer at -10 degrees C or in liquid nitrogen) should possibly be regarded as being inaccurate, unless the contrary can be demonstrated for different worm genera. However, exceptions are expected for the more rugged ova, such as those of the ascarids and Trichuris spp.  相似文献   

5.
SUMMARY Modifications of the McMaster egg counting technique are presented and discussed including the breaking up of hard faecal pellets, removal of air bubbles prior to counting, avoidance of salt spillage on the counting microscope and the use of sodium azide for preventing development of nematode eggs in field samples.  相似文献   

6.
A coprological sedimentation method is evaluated for quantification of egg shedding in bovine faeces. Through the inclusion of different number of Fasciola hepatica eggs in negative faeces, egg recovery rate and the sensitivity of the method were determined. The mean egg recovery rate of the technique was 76.72+/-15.42%. The sensitivity of the method was 33.3% whenever eggs/g of faeces (EPG) are less than 1.5 and 100% for higher values. To improve the diagnostic accuracy with this technique, it is advisable to increase the sample from 10 to 30g of faeces when manipulating low egg shedding, which allowed for a sensitivity of 83.3%. Regression equations were calculated to quantify the relationship between the number of recovered and incorporated eggs.  相似文献   

7.
The use of fecal egg count techniques to indirectly assess intestinal parasite burdens and determine anthelmintic efficacy is common in parasitological research and veterinary practice. The McMaster method is one of the most widely used techniques in veterinary practice, but recently, the Mini-FLOTAC technique has been introduced as a possible alternative. Studies comparing the two methods in precision and accuracy are needed. This study aimed at evaluating the Mini-FLOTAC technique for determining equine strongyle egg counts through a two-part procedure. First, a set of fecal counts was executed using both methods. Next, blind counts were performed on spiked fecal samples with true egg counts at 0, 5, 50, 500, and 1,000 eggs per gram. All counts were performed in triplicates, and each sample was counted using both methods. Mini-FLOTAC and McMaster had 83.2% and 53.7% precision, respectively. The accuracy was found to be 42.6% and 23.5% for Mini-FLOTAC and McMaster, respectively. In conclusion, this study found that Mini-FLOTAC exhibited both higher precision and accuracy than the McMaster technique and appears to be a more reliable alternative. Using a more precise egg-counting method can help assure that changes in egg counts before and after treatment reflect a genuine reduction and are not due to chance variability.  相似文献   

8.
The comparative efficacies of seven published McMaster method modifications for faecal egg counting were evaluated on pig faecal samples containing Ascaris suum eggs. Comparisons were made as to the number of samples found to be positive by each of the methods, the total egg counts per gram (EPG) of faeces, the variations in EPG obtained in the samples examined, and the ease of use of each of the methods. Each method was evaluated after the examination of 30 samples of faeces. The positive samples were identified by counting A. suum eggs in one, two and three sections of newly designed McMaster chamber. In the present study compared methods were reported by: I-Henriksen and Aagaard [Henriksen, S.A., Aagaard, K.A., 1976. A simple flotation and McMaster method. Nord. Vet. Med. 28, 392-397]; II-Kassai [Kassai, T., 1999. Veterinary Helminthology. Butterworth-Heinemann, Oxford, 260 pp.]; III and IV-Urquhart et al. [Urquhart, G.M., Armour, J., Duncan, J.L., Dunn, A.M., Jennings, F.W., 1996. Veterinary Parasitology, 2nd ed. Blackwell Science Ltd., Oxford, UK, 307 pp.] (centrifugation and non-centrifugation methods); V and VI-Gr?nvold [Gr?nvold, J., 1991. Laboratory diagnoses of helminths common routine methods used in Denmark. In: Nansen, P., Gr?nvold, J., Bj?rn, H. (Eds.), Seminars on Parasitic Problems in Farm Animals Related to Fodder Production and Management. The Estonian Academy of Sciences, Tartu, Estonia, pp. 47-48] (salt solution, and salt and glucose solution); VII-Thienpont et al. [Thienpont, D., Rochette, F., Vanparijs, O.F.J., 1986. Diagnosing Helminthiasis by Coprological Examination. Coprological Examination, 2nd ed. Janssen Research Foundation, Beerse, Belgium, 205 pp.]. The number of positive samples by examining single section ranged from 98.9% (method I), to 51.1% (method VII). Only with methods I and II, there was a 100% positivity in two out of three of the chambers examined, and FEC obtained using these methods were significantly (p<0.01) higher comparing to remaining methods. Mean FEC varied between 243 EPG (method I) and 82 EPG (method IV). Examination of all three chambers resulted in four methods (I, II, V and VI) having 100% sensitivity, while method VII had the lowest 83.3% sensitivity. Mean FEC in this case varied between 239 EPG (method I) and 81 EPG (method IV). Based on the mean FEC for two chambers, an efficiency coefficient (EF) was calculated and equated to 1 for the highest egg count (method I) and 0.87, 0.57, 0.34, 0.53, 0.49 and 0.50 for remaining methods (II-VII), respectively. Efficiency coefficients make it possible not only to recalculate and unify results of faeces examination obtained by any method but also to interpret coproscopical examinations by other authors. Method VII was the easiest and quickest but least sensitive, and method I the most complex but most sensitive. Examining two or three sections of the McMaster chamber resulted in increased sensitivity for all methods.  相似文献   

9.
A survey was carried out to determine the prevalence and seasonal abundance of the egg and adult stages of nematode parasites of sheep and goats in the semi-arid zone of north-eastern Nigeria between January and December 2002. Faecal samples collected from 102 sheep and 147 goats and examined by the modified McMaster technique using saturated solution of sodium chloride as the floating medium revealed that 44 (43.1%) and 82 (55.8%) of the samples, respectively, contained at least one nematode egg type. Three nematode egg types were recovered with strongyle egg type (22.5% in sheep and 35.4% in goats) being the most prevalent followed, respectively, by Trichuris (5.9% in sheep and 4.1% in goats) and Strongyloides (4.9% in sheep and 4.1% in goats) egg types. Mean faecal egg counts were generally moderate in both sheep (1052+/-922 strongyle, 1000+/-590 Strongyloides and 380+/-110 Trichuris eggs, respectively, per g of faeces) and goats (2092+/-3475 strongyle, 958+/-854 Strongyloides and 683+/-512 Trichuris eggs, respectively, per g of faeces) and showed the same trend irrespective of the age or sex of the animals. The prevalence and counts of strongyle nematode eggs showed a definite seasonal sequence that corresponded with the rainfall pattern in the study area during the period. In both sheep and goats, counts of strongyle egg type increased with the rains and reached peak levels at about the peak of the rainy season in September. The other egg types encountered during the study did not show much variation with the season of the year. Out of the 45 sheep and 75 goats examined at necropsy, 27 (60%) and 39 (52%), respectively, contained adult nematode species. Seven genera of adult nematodes including Strongyloides, Trichostrongylus, Haemonchus, Trichuris, Cooperia, Oesophagostomum and Bunostomum species were encountered during the study. Bunostomum species were recorded only in sheep. Adult worm burdens were generally low and showed seasonal variation that corresponded with the rainfall pattern in the study area during the period. Haemonchus and Trichostrongylus species attained peak counts together in both goats (June) and sheep (August). Strongyloides species were encountered throughout the year in both sheep and goats irrespective of the season. Other genera of nematodes encountered occurred in very low numbers and did not allow any meaningful comparison of seasonal sequence. The results suggest that Haemonchus, Trichostrongylus and Strongyloides species may be the major contributors to small ruminant helminthiasis in the study area.  相似文献   

10.
For the assessment of coccidial oocyst production by chickens, some modified form of the McMaster counting method is commonly used. The objective of this study was to evaluate a standard method and to compare it to a new, faster method, in which all the preparative stages before counting are carried out in the same container into which the original faecal sample was collected. A stock suspension containing purified oocysts of all seven valid Eimeria species that parasitize chickens was prepared, from which seven concentrations of oocyst suspensions were made. Since the faecal material in a sample influences the ability of oocysts to float up in a McMaster chamber, the new method was tested to establish the optimal amount of faeces in the original sample. Control oocyst suspensions containing no faeces were also tested, and three series of counts using the new method were compared with the standard McMaster method. The results were statistically analysed by agreement analysis. Repeatability and between-operator variation of both methods were also tested by agreement analysis. Counting by the standard McMaster method underestimated the true number of oocysts. The new method gave counts in agreement with the true number of oocysts if using 1 g of faeces per sample. With 2 g of faeces, counts were obtained that agreed with counts by the standard McMaster method. Both methods showed agreement between repeated measurements. The new method used on a sample containing 2 g of faeces provides a convenient alternative to the standard modified McMaster method. A 1-g faecal sample increases agreement with the true numbers of oocysts. Processing of a sample with the new method is about nine times faster than with the standard method.  相似文献   

11.
Haemonchus contortus eggs were extracted from sheep feces and known numbers were added to helminthologically sterile bovine feces to provide samples with seven, 30 and 60 eggs per gram (epg). At 60 epg, dilution techniques (modified Cornell-McMaster and modified McMaster) tended to overestimate the number of eggs and more eggs were recovered (mean of 121 and 88% respectively) with these techniques than with centrifugal concentration procedures (modified Cornell—63% and Wisconsin— 69%). At 30 epg, all techniques were comparable (modified Cornell-McMaster 67%, modified McMaster 63%, modified Cornell and Wisconsin 64%). At 7 epg, the Wisconsin (61%), modified Cornell (60%) and Cornell-McMaster (94%) techniques were comparable and better than the modified McMaster technique (16%). At all levels of epg, the modified Cornell and Wisconsin techniques recovered eggs from 100% of the samples. The Cornell-McMaster and modified McMaster techniques recovered eggs from 90 and 100% of samples at 60 epg; 40 and 100% at 30 epg; and 21 and 11% at 7 epg. With a gravitational concentration procedure, the Standard Vial, no more than 16% of the eggs at any level of epg were recovered and at 7 epg eggs were recovered from only one-half of the samples. Five gravitational concentration techniques were assessed over 66 to 490 epg. The Ovassay, Fecalyzer and modified Standard Vial techniques were comparable in efficiency (28%, 25% and 24% respectively), but the Standard Vial technique was less efficient (11%).

Introduced into diagnostic parasitology was the concept of predictive values which is the proportion of samples that a technique correctly identifies as being negative for parasite eggs. At 7 epg this was calculated to be zero for the modified Cornell-McMaster, modified McMaster and Standard Vial techniques and 100 for the Wisconsin and modified Cornell techniques.

  相似文献   

12.
Intestinal infections with Toxocara cati and Toxocara canis in their definitive host (felids and canids, respectively) are diagnosed by egg identification in faeces using coproscopical techniques. The Toxocara species is assumed to comply with the species from which the examined faeces were obtained, i.e. T. cati in cats and T. canis in dogs. We isolated and measured Toxocara eggs from faecal samples of 36 cats and 35 dogs from Switzerland and identified the Toxocara species by PCR. Amongst the isolates originating from dogs, 24 (68.5%) were determined as T. canis and 11 (31.5%) as T. cati. In all samples originating from cats, only T. cati was identified. Based on PCR identification, eggs of T. canis (n=241) and T. cati (n=442) were measured, revealing statistically significant different (p<0.001) mean sizes of 62.3 by 72.7 μm for T. cati and 74.8 by 86.0 μm for T. canis eggs. Considering that coprophagy is not unusual for dogs, a considerable percentage of Toxocara infections coproscopically diagnosed in dogs, as well as assumptions on anthelminthic resistance in regularly treated dogs, might in fact relate to intestinal passages of eggs following the uptake of other animals' faeces.  相似文献   

13.
Counts of Fasciola spp. eggs in faeces and measurements of antibody concentration to the excretory/secretory antigens of Fasciola spp. by ELISA were related to the numbers of flukes in the livers of 92 cattle killed in the abattoirs of Hanoi City, Vietnam. In this population, about 22% of the cattle had no flukes, another 22% had between 1 and 10 flukes, 44% between 11 and 100 flukes and 12% had more than 100 flukes in their livers. Of the 14 animals less than 2 years of age, only three were infected. At 2 years of age the mean number of flukes per liver was 10 whereas at 3 years and older, the mean varied between 60 and 80 flukes. Prevalence of infection was 78.3%. No eggs of Fasciola spp. were detected in the faeces of one third of infected cattle and 60% of the counts were less than 100 eggs per gram. The sensitivity of the egg counting method was 66.7% and specificity 100%, overall accuracy was 73.9%. Corresponding values for the ELISA method were 86.1, 70 and 82.6%, respectively. The positive and negative predictive values for the egg counting method were 100 and 45.5% and for the ELISA method were 91.2 and 58.3%, respectively.  相似文献   

14.
The conventional method for estimating the average strongyle egg count for a group of sheep was compared with a single count from a group composite faecal sample. Sixty-one groups of field samples were used. Composite samples were prepared in the laboratory by pooling equal amounts of faeces from individual samples. Data were logtransformed for analysis to meet the assumption of normality. There were no significant differences in the variances and overall mean counts obtained by the 2 methods. The regression line of log (composite count) on log (group arithmetic mean) did not differ significantly from the line of identity. When untransformed egg count data were categorised as low, moderate and high, the 2 methods were in agreement for 53 of the 61 groups. The mixing and counting process used for both methods (modified McMaster technique) gave highly repeatable results (repeatability = 0.94). The composite method was a quicker and valid alternative to the conventional method for monitoring helminthosis in sheep flocks.  相似文献   

15.
Previous observations showed that Duddingtonia flagrans chlamydospores were visualized in McMaster chambers containing faeces of treated sheep. This trial explored the McMaster technique as a tool to quantify chlamydospores in sheep faeces. A range of individual chlamydospore doses (from 19.5 x 10(6) to 177.5 x 10(6)) were offered orally to nine lambs for 7 consecutive days. A faecal sample (5 g) was daily obtained from the rectum of each animal (from days 1 to 13) to perform the McMaster technique using a sugar flotation fluid with 1.27 g/mL density. Each chlamydospore counted in the McMaster chamber was considered as 50 chlamydospores per g of faeces (CPG). The results confirmed that the estimated CPG was associated with the daily dose offered to the animals (r(2)=0.90; P<0.001). Furthermore, the total chlamydospore dose received by each animal was strongly associated to the total quantity of CPG obtained from the bulk faeces (TCtot) (r(2)=0.96; P<0.0001). Quantification of CPG can be used as a helpful tool to determine the number of chlamydospores reaching the faeces in orally dosed animals. This could be used to evaluate the efficacy of D. flagrans for the control of gastrointestinal nematode larvae in sheep faeces.  相似文献   

16.
The importance of preparation technique, culture media and incubation time in the embryonation of the infective egg stages of the intestinal nematode parasite Heterakis gallinarum was studied. Mature H. gallinarum worms were isolated from the caeca of infected chickens and separated by sex. In a first experiment intact female worms were kept for the development of their eggs in four different media (0.5% formalin, 2% formalin, 0.1 N sulphuric acid, 0.1% potassium dichromate) and incubated under constant temperature (20-22 degrees C) for 2, 4, 6 or 8 weeks. Afterwards the body of the worms were ruptured and the numbers of unembryonated and embryonated eggs were determined using a McMaster egg counting chamber, and the percentage of embryonated eggs was calculated. After 8 weeks of incubation in 0.5% formalin, 0.1 N sulphuric acid or 0.1% potassium dichromate 27.6%, 26.7% and 29.4% of the eggs, respectively, embryonated into third stage larvae (p > 0.05). In contrast, incubation in 2% formalin resulted in an embryonation of 18.6% only (p < 0.05). In a second experiment H. gallinarum eggs were directly harvested from worm uteri and cultivated afterwards in different media (2% formalin, 0.1 N sulphuric acid, 0.1% potassium dichromate) at 20 to 22 degrees C for 6 weeks. An incubation of isolated eggs in 2.0% formalin or 0.1% potassium dichromate during 6 weeks resulted in a significantly higher percentage of embryonation in comparison to the incubation of intact worms (first experiment). The results suggest that preparation technique, media and time of incubation has an essential influence on the development rate of H. gallinarum eggs.  相似文献   

17.
The faecal egg count (FEC) is the most widely used means of quantifying the nematode burden of horses, and is frequently used in clinical practice to inform treatment and prevention. The statistical process underlying the FEC is complex, comprising a Poisson counting error process for each sample, compounded with an underlying continuous distribution of means between samples. Being able to quantify the sources of variability contributing to this distribution of means is a necessary step towards providing estimates of statistical power for future FEC and FECRT studies, and may help to improve the usefulness of the FEC technique by identifying and minimising unwanted sources of variability. Obtaining such estimates require a hierarchical statistical model coupled with repeated FEC observations from a single animal over a short period of time. Here, we use this approach to provide the first comparative estimate of multiple sources of within-horse FEC variability. The results demonstrate that a substantial proportion of the observed variation in FEC between horses occurs as a result of variation in FEC within an animal, with the major sources being aggregation of eggs within faeces and variation in egg concentration between faecal piles. The McMaster procedure itself is associated with a comparatively small coefficient of variation, and is therefore highly repeatable when a sufficiently large number of eggs are observed to reduce the error associated with the counting process. We conclude that the variation between samples taken from the same animal is substantial, but can be reduced through the use of larger homogenised faecal samples. Estimates are provided for the coefficient of variation (cv) associated with each within animal source of variability in observed FEC, allowing the usefulness of individual FEC to be quantified, and providing a basis for future FEC and FECRT studies.  相似文献   

18.
Measurements of parasite load are often very variable. This implies that little confidence can be attached to single measurements of parasite numbers and egg concentrations, and that many measurements are required for the detection of differences between groups of hosts or parasites. For studies that aim to detect these differences, it is important to increase the precision (closeness of repeated measures to each other) of parasite numbers, because it determines the number of samples that is needed to find significant differences among groups. In this study, sample sizes required to detect group differences were estimated using nematode egg counts of faecal samples of dairy cattle. They were found to be much lower for a centrifugation technique than for the widely used McMaster technique in replicate samples, in spite of a generally similar mean FEC. For example, the sample size required to detect FEC differences between groups of 10, 50, and 250 eggs per gram (EPG) were 46, 25, and 27 for the McMaster technique and 8, 5, and 12 for the SSF method, respectively. Interestingly, sample sizes required for faeces with a relatively high egg concentration (approximately 1000 EPG) were also considerably lower than for the McMaster technique in spite of a higher mean EPG of the latter method. This implies that technical variation can be reduced considerably by simple methods of egg isolation. Given that the range of egg concentration is similar for a number of nematodes of livestock and human helminths, a reduction of technical error will aid studies with many group comparisons such as vaccination strategies against parasites with typically low FECs and studies of the genetics of host resistance. It may also lead to improved guidelines for measures related to public health.  相似文献   

19.
A multiplex PCR assay for differentiating strongyle eggs from cattle has recently been described; however, the egg disruption and DNA extraction procedures, though effective, are inadequate for large studies or clinical application. The purpose of this research was to evaluate methods for disrupting trichostrongyle eggs, then assess commercial kits for extracting egg DNA using Ostertagia ostertagi as a model species. Egg disruption procedures tested included probe sonication, bath sonication, bead beating, boiling, microwaving, proteinase K/SDS digestion, freezing, and various combinations of the above with the incorporation of sodium dodecyl sulfate. These procedures were evaluated in conjunction with four commercial DNA extraction kits: DNA Stool mini kit and DNeasy Plant kit (Qiagen), Fast DNA kit (QBiogene), and the MAP extraction kit (Tetracore). Results showed that egg disruption was best accomplished with the bead beater and ceramic beads, resulting in 100% disruption within 1min. When DNA extraction was preceded by the isolation of eggs from feces, all procedures except the Fast DNA kit produced PCR-ready DNA from at least two eggs. The DNeasy Plant kit allowed consistent detection of DNA released from one egg. Due to the morphological similarities among trichostrongyle eggs in ruminants, strongyle eggs in equids, and hookworm eggs, the methods described herein may have broad application to other nematodes.  相似文献   

20.
家蚕一代杂交种的良卵数、良卵率是蚕种品质检验的2项重要指标。为了提高检验效率,利用ImageJ图像处理软件建立检测家蚕一代杂交种良卵数、良卵率的新方法。分别利用这种图像处理方法和人工计数方法检测12个家蚕品种的蚕卵样本每克蚕卵的良卵数、总卵数,采用新方法检测获得的结果数据的相对误差均<1%。对2种方法获取的每克蚕卵良卵数、总卵数数据进行t测验,其t值均小于t0.05(22)=2.074,说明2种检测方法获取的检测结果差异不显著。相关性分析表明,利用2种检测方法对每克蚕卵良卵数、总卵数的检测结果的相关系数分别为0.9979、0.9982,2种检测方法获得的结果数据间呈极显著的线性相关。与人工计数检测方法相比,采用ImageJ图像处理技术的检测方法具有方便快捷的特点,可用于生产上家蚕一代杂交种的良卵数、良卵率检测。  相似文献   

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