The bias, accuracy and precision of faecal egg count reduction test results in cattle using McMaster, Cornell-Wisconsin and FLOTAC egg counting methods

The faecal egg count reduction test (FECRT) is the recommended method to monitor anthelmintic drug efficacy in cattle. There is a large variation in faecal egg count (FEC) methods applied to determine FECRT. However, it remains unclear whether FEC methods with an equal analytic sensitivity, but with different methodologies, result in equal FECRT results. We therefore, compared the bias, accuracy and precision of FECRT results for Cornell-Wisconsin (analytic sensitivity = 1 egg per gram faeces (EPG)), FLOTAC (analytic sensitivity = 1 EPG) and McMaster method (analytic sensitivity = 10 EPG) across four levels of egg excretion (1-49 EPG; 50-149 EPG; 150-299 EPG; 300-600 EPG). Finally, we assessed the sensitivity of the FEC methods to detect a truly reduced efficacy. To this end, two different criteria were used to define reduced efficacy based on FECR, including those described in the WAAVP guidelines (FECRT <95% and lower limit of 95%CI <90%) (Coles et al., 1992) and those proposed by El-Abdellati et al. (2010) (upper limit of 95%CI <95%). There was no significant difference in bias and accuracy of FECRT results across the three methods. FLOTAC provided the most precise FECRT results. Cornell-Wisconsin and McMaster gave similar imprecise results. FECRT were significantly underestimated when baseline FEC were low and drugs were more efficacious. For all FEC methods, precision and accuracy of the FECRT improved as egg excretion increased, this effect was greatest for McMaster and least for Cornell-Wisconsin. The sensitivity of the three methods to detect a truly reduced efficacy was high (>90%). Yet, the sensitivity of McMaster and Cornell-Wisconsin may drop when drugs only show sub-optimal efficacy. Overall, the study indicates that the precision of FECRT is affected by the methodology of FEC, and that the level of egg excretion should be considered in the final interpretation of the FECRT. However, more comprehensive studies are required to provide more insights into the complex interplay of factors inherent to study design (sample size and FEC method) and host-parasite interactions (level of egg excretion and aggregation across the host population).     

Effects of aggregation and sample size on composite faecal egg counts in sheep

Composite faecal egg counts (FEC) are increasingly used to support strategic anthelmintic treatment decisions in grazing livestock. However, their accuracy as estimators of group mean FEC is affected by the number of individual samples included, how thoroughly they are mixed, and the underlying degree of parasite aggregation between individual hosts. This paper uses a Negative Binomial model for parasite aggregation, and a Poisson model for egg distribution within faecal suspensions, in order to optimise composite FEC protocol for commercial sheep flocks. Our results suggest that faecal egg density in a well-mixed composite sample from 10 sheep (3g of faeces from each), estimated by examination of four independently filled McMaster chambers, is likely to provide an adequate estimate of group mean FEC in the majority of situations. However, extra care is needed in groups of sheep for which high levels of FEC aggregation might be expected. The implications of statistical error in FEC estimates depend on how they are used. The simulation-based approach presented here is a powerful tool for investigating the risks of error in FEC-driven treatment decisions in different situations, as well as for the statistical analysis of parasitological data in general.     

Calibration and diagnostic accuracy of simple flotation, McMaster and FLOTAC for parasite egg counts in sheep

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.     

Novel insights in the faecal egg count reduction test for monitoring drug efficacy against gastrointestinal nematodes of veterinary importance

The faecal egg count reduction test (FECRT) is the method of choice to monitor anthelmintic efficacy against gastro-intestinal nematodes in livestock. Guidelines on how to conduct a FECRT are made available by the World Association for the Advancement of Veterinary Parasitology (WAAVP). Since the publication of these guidelines in the early 1990 s, some limitations have been noted, including (i) the ignorance of host-parasite interactions that depend on animal and parasite species, (ii) their feasibility under field conditions, (iii) appropriateness of study design, and (iv) the high detection limit of the recommended faecal egg count (FEC) method. Therefore, the objective of the present study was to empirically assess the impact of the level of excretion and aggregation of FEC, sample size and detection limit of the FEC method on the sensitivity and specificity of the FECRT to detect reduced efficacy (<90% or <95%) and to develop recommendations for surveys on anthelmintic resistance. A simulation study was performed in which the FECRT (based on the arithmetic mean of grouped FEC of the same animals before and after drug administration) was conducted under varying conditions of mean FEC, aggregation of FEC (inversely correlated with k), sample size, detection limit and 'true' drug efficacies. Classification trees were built to explore the impact of the above factors on the sensitivity and specificity of detecting a truly reduced efficacy. For a reduced-efficacy threshold of 90%, most combinations resulted in a reliable detection of reduced and normal efficacy. For the reduced-efficacy threshold of 95% however, unreliable FECRT results were found when sample sizes <15 were combined with highly aggregated FEC (k=0.25) and detection limits ≥ 5 EPG or when combined with detection limits ≥ 15 EPG. Overall, an increase in sample size and mean preDA FEC, and a decrease in detection limit improved the diagnostic accuracy. FECRT remained inconclusive under any evaluated condition for drug efficacies ranging from 87.5% to 92.5% for a reduced-efficacy-threshold of 90% and from 92.5% to 97.5% for a threshold of 95%. The results highlight that (i) the interpretation of this FECRT is affected by a complex interplay of factors, including the level of excretion and aggregation of FEC and (ii) the diagnostic value of FECRT to detect small reductions in efficacy is limited. This study, therefore, provides a framework allowing researchers to adapt their study design according to a wide range of field conditions, while ensuring a good diagnostic performance of the FECRT.     

A comparison of modifications of the McMaster method for the enumeration of Ascaris suum eggs in pig faecal samples

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.     

Comparison between methods for measuring fecal egg count and estimating genetic parameters for gastrointestinal parasite resistance traits in sheep

Fecal egg count (FEC) is an indicative measurement for parasite infection in sheep. Different FEC methods may show inconsistent results. Not accounting for inconsistencies can be problematic when integrating measurements from different FEC methods for genetic evaluation. The objectives of this study were to evaluate the difference in means and variances between two fecal egg counting methods used in sheep—the Modified McMaster (LMMR) and the Triple Chamber McMaster (LTCM); to estimate variance components for the two FEC methods, treating them as two different traits; and to integrate FEC data from the two different methods and estimate genetic parameters for FEC and other gastrointestinal parasite resistance traits. Fecal samples were collected from a commercial Rideau-Arcott sheep farm in Ontario. Fecal egg counting was performed using both LMMR and the LTCM methods. Other parasite resistance trait records were collected from the same farm including eye score (FAMACHA), body condition score (BCS), and body weight (WT). The two FEC methods were highly genetically (0.94) and phenotypically (0.88) correlated. However, the mean and variance between the two FEC methods were significantly different (P < 0.0001). Therefore, re-scaling is required prior to integrating data from the different methods. For the multiple trait analysis, data from the two fecal egg counting methods were integrated (LFEC) by using records for the LMMR when available and replacing missing records with re-standardized LTCM records converted to the same mean and variance of LMMR. Heritability estimates were 0.12 ± 0.04, 0.07 ± 0.05, 0.17 ± 0.06, and 0.24 ± 0.07 for LFEC egg count, FAMACHA, BCS, and WT, respectively. The estimated genetic correlations between FEC and the other parasite resistance traits were low and not significant (P > 0.05) for FAMACHA (r = 0.24 ± 0.32) and WT (r = 0.22 ± 0.19), and essentially zero for BCS (r = −0.03 ± 0.25), suggesting little to no benefit of using such traits as indicators for LFEC.     

Gastrointestinal nematode burden in working equids from humid tropical areas of central Veracruz, Mexico, and its relationship with body condition and haematological values

The east coast of Veracruz, Mexico, has an important equine population used for working in rural production systems. The objectives of this study were (1) to calculate the prevalence of tropical working equids (donkeys, mules and horses) infected with gastrointestinal nematodes (GINs) and the GINs involved, and (2) to measure the body condition score (BCS) and haematological values for each working equid and its relationship with faecal worm egg count (EPG). One hundred and forty working equids were randomly selected from five different villages along the central coast of the state of Veracruz and faecal and blood samples were obtained from each animal. Gastrointestinal parasite burdens were determined using the McMaster technique. Packed cell volume, total plasma proteins, red blood cell count and white blood cell count were measured from each blood sample. Prevalence of infected equids was higher than 90 %. Mules had the highest median faecal worm egg counts (875 EPG), followed by horses and donkeys with 400 EPG. There was no correlation between EPG and BCS or haematological values (p?>?0.05). Results suggest that despite the high prevalence and parasite burdens, equids involved in this trial are not being seriously affected. This study provides information which might help in designing future strategies to control nematode infections in working equids in the Mexican tropics; more emphasis should be placed on other inputs (nutrition perhaps), with individual anthelminthic treatment to those animals with the highest EPG or when signs present themselves.     

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 strongyles and Dicrocoelium dendriticum in sheep

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.     

The efficacy and acceptability of ivermectin liquid compared to that of oral paste in horses

One hundred-twenty horses and ponies ranging in age from 142 days to 23 years were used to assess the efficacy and acceptability of ivermectin liquid for horses when given as an oral drench or by nasogastric intubation. Prior to treatment, animals in this study were found to have eggs in the feces of one or more of the following: strongyle type, Parascaris equorum, and Strongyloides westeri. While egg parasite per gram (EPG) numbers from 30 untreated controls remained consistently positive over a 14 day period, parasite EPG numbers from animals treated on Day 0 were reduced to 0 by day 14 as determined by a modified McMaster technique.     

Prevalence of common gastrointestinal nematode parasites in scavenging pigs of different ages and sexes in eastern centre province, Burkina Faso

The range and infestation intensities of gastrointestinal parasitic nematode species depend on the type of swine production system. The present study focused mainly on nematodes of veterinary importance in scavenging pigs in Burkina Faso, and aimed at determining the prevalence of gastro-intestinal nematode parasites by means of faecal egg per gram (EPG) counts. Between November 2001 and October 2002, faecal samples from 383 pigs of different sexes and ages (< 5 months, 5-12 months and > 12 months) were collected from the rectum and examined for gastrointestinal nematodes parasites using the Mc Master method. Of the 383 pigs examined, 91% were infected by one or more parasites. Ascaris suum (40%; 100-1 400 EPG) was the most prevalent parasite followed by Strongyloides ransomi (21%; 100-4200 EPG), Oesophagostomum spp. (18%; 100-1000 EPG), Hyostrongylus rubidus (11%; 100-1 800 EPG), Globocephalus spp. (10%; 100-400 EPG) and Trichuris suis (1 %; 100-200 EPG). The prevalence was significantly higher in female pigs (n = 239) than in males. In addition, females excreted significantly (P < 0.05) more eggs in their faeces than males, except in the case of Globocephalus spp. The age of the animal had no effect on the prevalence of A. suum whereas there were significant differences in age categories concerning S. ransomi, H. rubidus, Oesophagostumum spp. and Globocephalus spp. Unexpectedly, the high prevalence of these common parasites was not accompanied by elevated EPG values, which suggests the existence of moderate infestations. The present work indicates that the common nematode infestations in pigs do not necessarily need a systematic herd anthelmintic treatment, as only a small number of worms is required to induce immunity. A further study is needed to formulate appropriate and cost-effective strategies for the control of gastro-intestinal nematode parasites in pigs in Burkina Faso.     

Gastrointestinal parasites presence during the peripartum decreases total milk production in grazing dairy Holstein cows

Parasitism in cattle is known to impair growth and development. Recent findings suggest that productivity of adult animals is also affected, but little is known about the physiological mechanisms involved. Furthermore, development of nematode resistance to drugs makes imperative the search of management practices that avoid whole herd treatment. We undertook an epidemiological and endocrine study in a grass based dairy farm in Argentina to study the effect of parasites on milk production and the underlying mechanisms involved, and identify individual animals that would benefit from antiparasitic treatment. All the cows in the dairy were followed monthly for egg parasite output in feces. Samples were cultured for genera determination. Milk production and reproductive results were recorded and periodical bleedings for hormone determination were performed. Nematode egg output (EPG) was maximal in late Summer and Autumn and minimal in Spring in coincidence with the Ostertagia inhibition-disinhibition cycle as this genus had the highest prevalence in all the study. The highest proportion of positive samples was found in the high producing herd and maximal counts were found in the peripartal period. Milk production did not correlate with EPG mean values but, when cows were grouped by EPG positivity around parturition, a significant difference in total milk production between EPG null and positive cows was observed. Positive cows produced 7%, 12% or 15% less milk than null EPG cows, depending on the sampling month/s chosen for classification. The highest difference was seen when both prepartum and postpartum samples were taken into account. No difference in lactation length and a marginal effect on partum to first service interval were encountered. Endocrine studies revealed a decrease in serum growth hormone (GH), type I insulin-like growth factor (IGF-I) and prolactin during lactation in cows with positive EPG in the first postpartum sample with respect to null EPG cows at that time. GH levels decreased and prolactin and IGF-I levels increased in both groups of cows from month 0 to 6 in milk. Serum insulin levels remained stable throughout lactation and were similar in both groups of cows. In conclusion, EPG around parturition may be a useful tool for identifying cows that will have a decrease in productivity due to parasite effects and would possibly benefit from an antiparasitic treatment. Besides, our results suggest that detrimental effect of parasites on milk production may be mediated by GH, IGF-I and prolactin serum levels.     

Impact on productivity of peri-parturient rise in fecal egg counts in Creole goats in the humid tropics

The control of gastrointestinal nematodes requires an understanding of their epidemiology so that particular parasite stages can be targeted. Dam infection during early lactation is one example of this in ruminant nematode infections. The existence of the peri-parturient relaxation in immunity and its impact on productivity were examined in a Creole goat flock from Guadeloupe, exposed to mixed natural infection (predominantly Haemonchus contortus and Trichostrongylus colubriformis). A total of 1,511 l were obtained from 909 does resulting from 463 dams and 150 sires. Fecal and blood samples were collected at kidding before anthelmintic drenching, 4 and 6 weeks after kidding. The traits analyzed were logarithm transformed fecal egg counts (FEC), packed cell volume (PCV), and logarithm transformed blood eosinophilia counts (EOS) for does at each sampling point and changes in these during the postpartum period. With the exception of the PCV values measured at kidding, lactating does had significantly higher FEC and lower PCV than control dry does at every sampling point. Geometric means of FEC in lactating does were 819 +/- 174, 677 +/- 146 and, 699 +/- 160 eggs per gram (EPG) at kidding, 4 and 6 weeks after kidding respectively. Geometric means of FEC in dry does were 187 +/- 57, 89 +/- 28, 133 +/- 43 at these time points, respectively. EOS differences were not consistent between groups and probably not specific enough for variations in Creole goats' peri-parturient rise to be discussed. As does aged, their egg output decreased and primiparous does always had greater egg output than multiparous ones. Overall, does' FEC at 4 weeks after kidding decreased by 1.3% each year. The higher the litter size, the higher the FEC at kidding and inverse applied for PCV measurements. Does that stopped lactating had significantly lower FEC and higher PCV values than lactating does with low milk yields. Higher infection rates during early lactation in Creole goats were recorded in does with lower maternal ability assessed by the average daily weight gain of kids between 10 and 30 days of age. Kids from dams with higher FEC (i.e. >600 EPG higher than corresponding does) had 17% lower average daily weight gain between 30 and 70 days postpartum and were approximately 1 kg lighter at weaning than kids from dams with lower FEC. Thus, it is clear that a peri-parturient rise in FEC exists in Creole goats. By controlling the intensity of this peri-parturient rise in FEC, herd health and productivity could be substantially improved.     

Quantifying the sources of variability in equine faecal egg counts: implications for improving the utility of the method

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.     

Precision, repeatability and representative ability of faecal egg counts in Heterakis gallinarum infected chickens

This study investigated whether a precise and repeatable quantification of Heterakis gallinarum egg excretion, which considerably reflects the actual worm burdens, can be achieved based on collection of the daily total amount of faeces from chickens. Three-week-old birds (N=64) were infected with 200 embryonated eggs of H. gallinarum, and placed into individual cages 3 wk after infection for 5 wk to collect daily faeces (N=2240). The total daily faeces was mixed and a randomly taken sample per bird was analyzed to estimate the numbers of eggs per gram of faeces (EPG) and total number of eggs excreted within 24h (EPD). A total of 235 daily faecal collections were randomly selected and further examined to determine between and within sample variations of EPG counts as a measure of precision. For this, two random faecal samples were taken from the daily produced faeces by a bird, and the EPG was determined for each of the samples (EPG1 and EPG2). The second faecal sample was analyzed once more to determine a parallel EPG2 count (EPG2a) of the suspended sample. Precision of an EPG count was defined as its relative closeness to the average of two EPG counts using a relative asymmetry index (Index(EPG)). At an age of 11 wk, i.e. 8 wk p.i. the birds were slaughtered and their worm burdens were determined. There were no significant differences between EPG1 and EPG2 (P=0.764) nor between EPG2 and EPG2a (P=0.700), suggesting that the differences between or within the samples were not different from zero. Correlations between EPG counts, as between and within sample coherences, were r=0.85 and r=0.86, respectively. Precision of EPG counts, as measured by Index(EPG), was not influenced by consistency (P=0.870) and total amount of faeces (P=0.088). However, concentration of eggs in faeces (mean EPG) had a significant effect on the precision of the EPG counts (P<0.001). Similar results were also observed for the within sample precision (Index(EPG2)). A segmented regression analysis indicated an abrupt change in the precision of EPG counts as the response to changing egg concentration in the examined faecal samples. The precision of analyses remarkably heightened up to a breakpoint with an EPG count of ≤ 617. A similar breakpoint was also determined for within sample precision (EPG2 ≤ 621). Moderate repeatabilities (R=0.49) for EPG and EPD were estimated in the first week of egg excretion, whereas the estimates were higher (R=0.67-0.84) in the following weeks. Correlations between number of female worms with daily measured EPG and EPD increased to an almost constant level (r ≥ 0.70; P<0.05) in a few days after the nematode excreted eggs and predominantly remained so for the rest of the sampling period. It is concluded that mixing daily total faeces provides samples with random homogenous distribution of H. gallinarum eggs. Precision of the EPG counts increases as the egg concentration in faecal sample increases. Egg excretion of H. gallinarum, quantified either as EPG or EPD, is highly repeatable and closely correlated with the actual worm burden of birds starting as early as in 5 th wk of infection.     

Comparison of methods to detect gastrointestinal parasites in llamas and alpacas

OBJECTIVE: To compare relative sensitivity and overall yields of various methods of fecal examination for gastrointestinal parasites in llamas and alpacas. DESIGN: Prospective study. SAMPLE POPULATION: Fecal samples from 42 alpacas and 62 llamas. PROCEDURES: Fecal samples were analyzed via direct smear, a modified McMaster technique with sucrose solution or saturated saline (approx 36% NaCl) solution, and a centrifugation-flotation procedure. McMaster flotation chambers were examined 15 and 60 minutes after loading. Centrifugation-flotation samples were examined after 10 and 60 minutes of flotation. The proportions of samples with positive results and concentrations of parasites were compared among methods. RESULTS: The centrifugation-flotation technique yielded more positive results than other methods for all parasites except small coccidia. Longer flotation time increased the proportion of positive results and parasite concentrations for all parasites except Nematodirus spp. Longer time in the McMaster chamber made little difference. By use of the modified McMaster technique, sucrose solution yielded more positive results for Trichuris spp, Eimeria macusaniensis, and strongyles, whereas saline solution yielded more positive results for Nematodirus spp and small coccidia. The saline solution McMaster test yielded more positive results for small coccidia than did most other methods, and the sucrose McMaster technique yielded more positive results for Trichuris spp. CONCLUSIONS AND CLINICAL RELEVANCE: The centrifugation-flotation technique appeared to offer clear advantages in detecting infection with E macusaniensis, Trichuris spp, Nematodirus spp, and capillarids. The saline McMaster technique appeared to offer an advantage in detecting small coccidia.     

Accuracy and Precision of Mini-FLOTAC and McMaster Techniques for Determining Equine Strongyle Egg Counts

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.     

福州动物园圈养鸟类肠道寄生虫调查

为调查福州动物园圈养鸟类消化道寄生虫感染情况,采集16种禽类31份粪便样品,应用饱和盐水漂浮法和自然水洗沉淀法收集虫卵,麦克斯特计数法对采集的粪便样本进行虫卵测定。检查结果显示,寄生虫感染阳性样品数有23份,占样品总数的74%。总共检出6种寄生虫感染,按感染例数从大到小排序为:球虫(9例)、吸虫(7例)、绦虫(6例)、线虫(5例)、鞭虫(4例)、纤毛虫(3例)。感染寄生虫的禽类中,孔雀的感染强度最高,EPG为9 150个/g;其次是巨嘴鸟,EPG为4 500个/g;第三是东方白鹳,EPG为3 300个/g。其他鸟类的EPG分布在0~2 100个/g。该调查结果为福州动物园圈养鸟类寄生虫病的防治以及制定科学的驱虫程序提供可靠的依据。     

The contribution of simple random sampling to observed variations in faecal egg counts

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.     

In vivo anthelmintic activity of Dorycnium rectum and grape seed extract against Ostertagia (Teladorsagia) circumcincta and Trichostrongylus colubriformis in sheep

AIM: To assess the in vivo anthelmintic activity of condensed tannins (CT) in the forage species Dorycnium rectum and Medicago sativa, and in an extract from grape (Vitus vinifera) seeds (GSE), against two species of parasite, Teladorsagia (Ostertagia) circumcincta and Trichostrongylus colubriformis, at different stages of their life cycle, in sheep that were parasite-na?ve or previously exposed to nematodes. METHODS: In Trial 1, a factorial treatment structure was used to compare faecal nematode egg counts (FEC) and worm burdens in 40 weaned Romney lambs fed either the CT-containing forage D. rectum (12% dry matter; DM) or M. sativa (lucerne; 0.2% DM). Twenty na?ve and 20 previously-exposed lambs were drenched free of parasites then reinfected with known species and numbers of parasites, and housed in pens indoors on a diet of lucerne pellets and chaffed hay. Groups of lambs (n=5 lambs per group) were fed one of the forages over one of two time periods within the parasite's life cycle. Six to nine days after the last feeding of fresh forages, faecal samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. In Trial 2, 12 Suffolk x Romney lambs were surgically implanted with an abomasal cannula and then housed indoors in metabolism crates. After infection with parasites, six lambs were infused continuously over a 14-day period with a commercially available CT GSE (96% DM, made up to 34 g/L in water); the remaining lambs were infused with water. During infusion, samples were collected for egg hatch and larval development assays. After infusion, samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. RESULTS: In Trial 1, there was a significant (p<0.001) difference in burdens of O. circumcincta between na?ve lambs and those previously exposed to parasites, but no other differences were recorded. In Trial 2, lambs infused with GSE had significantly (p<0.05) fewer T. colubriformis at slaughter and significantly (p<0.001) fewer eggs hatched in the egg hatch assay (EHA) than for lambs infused with water. Overall, the differences attributable to GSE were small in magnitude, being an 11% drop in egg hatch, and an 18% drop in numbers of adult T. colubriformis after 14 days of continuous infusion. No other differences were recorded. CONCLUSION: The results indicate that the in vivo anthelmintic activity of these CT sources is, at best, modest and is unlikely to be of any practical value. Further, these data emphasise that in vitro activity is an unreliable indicator of in vivo efficacy for CT-containing forages and extracts.     

人工养殖鹦鹉肠道寄生虫感染情况调查

本文采用粪便直接涂片、浮聚和麦克马斯特法,对采自广州市番禺区欢乐鸟场的17种共183个鹦鹉粪便样品进行了肠道寄生虫感染率反感染强度调查。先后进行了两次采样,检查结果显示：该鸟场养殖的鹦鹉总肠道寄生虫感染率为38．3％。其中吸虫感染比较普遍,感染率达19．1％,其次是绦虫,感染率为10．4％o,线虫和原生动物的感染率分别为9．3％和5．5％。感染强度EPG值随寄生虫和鹦鹉种的不同,在300到2400之间不等。调查中没有发现球虫感染,鉴于调查结果建议对鸟场阳性感染鹦鹉进行针对性驱虫。