Relative egg extraction efficiencies of manual and automated fecal egg count methods in equines.

The World Association for the Advancement of Veterinary Parasitology recently released new recommendations for the design of fecal egg count (FEC) reduction tests for livestock. These provide suggestions as to the number of animals to be sampled and the minimum number of eggs that must be counted to produce statistically meaningful results. One of the considerations for study design is the multiplication factor of the FEC method to be used; methods with lower multiplication factors require fewer animals to be sampled because they are presumed to count more eggs per test. However, multiplication factor is not the sole determinant of the number of eggs counted by any given method, since different techniques use very different sample extraction methodologies that could affect the number of eggs detected beyond just the amount of feces examined. In this light, we compared three commonly used manual FEC methods (mini-FLOTAC, McMaster and Wisconsin) and two automated methods (Imagyst and Parasight All-in-One) with respect to how many equine strongylid and ascarid eggs they counted in the same samples. McMaster and mini-FLOTAC (multiplication factors of 25x and 5x, respectively) produced the most accurate results of the methods tested but mini-FLOTAC counted approximately 5-times more eggs than McMaster. However, Wisconsin and Parasight (multiplication factor = 1x) counted 3-times more ova than mini-FLOTAC, which was less than the 5-fold difference in their multiplication factors. As a result, these tests perform with multiplication factors more akin to 1.6x relative to mini-FLOTAC. Imagyst, due to its unique sample preparation methodology, does not have a traditional multiplication factor but performed similarly to McMaster with respect to egg recovery.

Epidemiological study on factors influencing the occurrence of helminth eggs in horses in Germany based on sent-in diagnostic samples.

Gastrointestinal nematodes are ubiquitous parasites of grazing equines with Parascaris spp., and strongyles being the most relevant ones regarding the prevalence and potential disease severity. Despite their importance, epidemiological data regarding the presence and egg-shedding intensities of these parasites are scarce. Data from 1067 horse samples collected on German horse farms initially to compare diagnostic methods were used for epidemiological analyses. Due to its higher sensitivity, presence/absence data were based on a combined sedimentation/flotation technique while faecal egg counts were based on Mini-FLOTAC. For strongyles, 46.5% of the samples were positive and the median egg-shedding intensity was 40 (range 5-2590). In multivariate analyses, prevalence and egg-shedding intensity were significantly influenced by season, age group and sample type. The drug used for the last treatment and the number of foals on the yard only affected prevalence while the number of horses on the yard and sex were only significant for egg-shedding intensity. For Parascaris spp., a prevalence of 4.6% and a median egg-shedding intensity of 0 (range 5-905) were observed. In multivariate analyses, the age group, the time since the last anthelmintic treatment, presence and number of foals had significant effects on ascarid prevalence whereas egg-shedding intensity was significantly influenced by age group and season only. Parascaris occurred only on yards with foals, but with an increasing number of foals, Parascaris egg-shedding intensity decreased. Prevalence and egg-shedding intensity were influenced by different but partially overlapping variables for Parascaris and strongyles.

High prevalence of pathogenic Eimeria spp. and the main risk factors associated with infection in goats from a semiarid region of Northeastern Brazil.

This study aimed to analyze the epidemiology of infections caused by Eimeria spp. in goats bred in an extensive production system in Northeast Brazil. To this end, an epidemiological survey was performed, wherein 350 fecal samples were collected from goats ranging from 3 months to 8 years old across 30 farms. The number of oocysts per gram of feces (OPG) was counted using the Mini-FLOTAC method. Species were identified based on their morphological and morphometric characteristics. Frequencies between animal categories were assessed using the chi-square test and known risk factors after multivariate analysis. Eimeria oocysts were found in 99.4% (346/348) of samples and 100% (30/30) of the farms. The OPG was influenced by the age of the animals; therefore, young goats (up to 6 months old) eliminated more oocysts (2302.6 + 5454.8) than adult goats (881.7 + 1551.5). Three pathogenic species were identified, with two demonstrating a high prevalence: Eimeria arloingi (88.83%) and Eimeria christenseni (44.7%), in addition to Eimeria caprina (3.72%). In the study, the habit of ground feeding and the use of cisterns as water sources were identified as risk factors, with odds ratios of 4.04 and 9.82, respectively. However, cleaning facilities at shorter intervals was a protective factor (OR = 0.19). We concluded that there was a high prevalence of Eimeria infection in goats bred in extensive production systems, even those in semiarid regions. This high prevalence can be attributed to the congregation of animals in shaded areas and the substantial accumulation of fecal matter at night.

Current status of intestinal parasitosis among patients attending teaching hospitals in Zagazig district, Northeastern Egypt.

Almost 80% of health problems in the developing world are due to malnutrition and infectious diseases, which are mainly parasitic. Updated records on the prevalence of parasitic infections and the potential risk factors are essential to enhancing control strategies. Therefore, this study was conducted to evaluate the current situation of intestinal parasitism among patients attending teaching hospitals in Zagazig district, Northeastern Egypt. The study involved five hundred cases. They were all subjected to faecal examination using direct smear measure and two commercial faecal concentrators: Mini-Parasep® solvent-free and Mini-FLOTAC procedures. Mini-FLOTAC was performed with two solutions (FS2: saturated sodium chloride and FS7: zinc sulphate). The overall prevalence of intestinal parasitic infections was 56%. Different species were identified, like Giardia lamblia (12.6%), Entamoeba histolytica/dispar (10%), Ascaris lumbricoides (8.8%) and Hymenolepis nana (8.6%). Data analyses revealed a significant association of intestinal parasitism with different socio-demographic features of the participants. Our results showed a better diagnostic performance of Mini-Parasep® in the overall recovery of intestinal parasites. It was more accurate than Mini-FLOTAC in diagnosing both helminths and protozoan infections. Mini-FLOTAC (FS2) exhibited a higher sensitivity than FS7 for helminth recovery (74.6% vs 53.4%), while FS7 was more sensitive for protozoan infections (50.6% vs 43.8%). Intestinal parasitosis remains a challenging health problem in Zagazig city, wherever reliable diagnostic approaches are limited. Thus, our study has proposed the value of the commercial concentrators (Mini-Parasep® and Mini-FLOTAC) as alternative techniques for diagnosing a large variety of parasite species in resource-constrained settings.

Ascaris lumbricoides eggs or artefacts? A diagnostic conundrum.

Due to the presence of artefacts in stool samples, the copromicroscopic diagnosis of Ascaris lumbricoides is not always straightforward, particularly in the case of fertilized decorticated eggs. A total of 286 stool samples from 115 schoolchildren in India and 171 adult immigrants in Italy were screened for the presence of A. lumbricoides eggs by both Kato-Katz thick smear and Mini-FLOTAC. If the outer layer of A. lumbricoides eggs was absent, two aliquots of each stool sample were preserved: one for coproculture to identify larvae after development and one to compose a pool of stool for molecular analysis. A total of 64 stool samples (22.4%) were positive for A. lumbricoides using the Kato-Katz thick smear; 36 (56.3%) of these showed mammillated A. lumbricoides eggs, 25 (39.1%) showed elements resembling fertilized decorticated eggs, while three samples (4.7%) showed both mammillated and decorticated eggs. By Mini-FLOTAC, 39 stool samples (13.6%) were positive, while decorticated A. lumbricoides-like eggs were identified as artefacts. These results were confirmed by negative coprocultures and quantitative polymerase chain reaction. Mini-FLOTAC can be used for a reliable diagnosis of A. lumbricoides, thanks to the flotation and translation features which allow a clearer view, resulting in the correct identification of A. lumbricoides eggs.

The Kubic FLOTAC microscope (KFM): a new compact digital microscope for helminth egg counts.

The Kubic FLOTAC microscope (KFM) is a compact, low-cost, versatile and portable digital microscope designed to analyse fecal specimens prepared with Mini-FLOTAC or FLOTAC, in both field and laboratory settings. In this paper, we present the characteristics of the KFM along with its first validation for fecal egg count (FEC) of gastrointestinal nematodes (GINs) in cattle. For this latter purpose, a study was performed on 30 fecal samples from cattle experimentally infected by GINs to compare the performance of Mini-FLOTAC either using a traditional optical microscope (OM) or the KFM. The results of the comparison showed a substantial agreement (concordance correlation coefficient = 0.999), with a very low discrepancy (−0.425 ± 7.370) between the two microscopes. Moreover, the KFM captured images comparable with the view provided by the traditional OM. Therefore, the combination of sensitive, accurate, precise and standardized FEC techniques, as the Mini-FLOTAC, with a reliable automated system, will permit the real-time observation and quantification of parasitic structures, thanks also to artificial intelligence software, that is under development. For these reasons, the KFM is a promising tool for an accurate and efficient FEC to improve parasite diagnosis and to assist new generations of operators in veterinary and public health.

Comparative survey of gastrointestinal parasites in sympatric Iberian ibex (Capra pyrenaica) and domestic goats using molecular host specific identification.

An isolated population of several hundred Iberian ibex (Capra pyrenaica) inhabits the Gerês-Xurés Transboundary Biosphere Reserve (GXTBR) in north-western Iberian Peninsula, in partial sympatry with tens of thousands domestic goats (Capra hircus). This study aimed to assess the prevalence and shedding intensity of gastrointestinal parasites from sympatric herds of domestic goat and Iberian ibex in autumn 2018. A total of 93 pooled faecal samples were collected from GXTBR (39 from domestic goats and 54 from Iberian ibex) and the host species was identified using molecular techniques, whenever defecation was not visualised in the field. Parasitological analysis was achieved by joint Willis flotation/sedimentation, McMaster and Mini-FLOTAC techniques. Seventy-two samples (25 domestic goats and 47 Iberian ibex) were retained for analysis after discarding the least fresh and those with uncertain specific identification. Generalized linear mixed models compared prevalence and shedding intensity between caprine species. Domestic goats showed a non-significant tendency to be more parasitized than Iberian ibex, as assessed by overall prevalence (100.0%, CI95 86.7-100.0 vs. 74.5%, CI95 69.5-84.8), and shedding intensity. This study reveals a similar community of gastrointestinal parasites in an abundant livestock species and an isolated population of wild caprine, living in partial sympatry. It is the first study on the health status of this endangered Iberian ibex population, in close contact with livestock, highlighting the need for further sanitary surveillance.

Preliminary evaluation of a novel, fully automated, Telenostic device for rapid field-diagnosis of cattle parasites.

New ideas for diagnostics in clinical parasitology are needed to overcome some of the difficulties experienced in the widespread adoption of detection methods for gastrointestinal parasites in livestock. Here we provide an initial evaluation of the performance of a newly developed automated device (Telenostic) to identify and quantify parasitic elements in fecal samples. This study compared the Telenostic device with the McMaster and Mini-FLOTAC for counting of strongyle eggs in a fecal sample. Three bovine fecal samples were examined, in triplicate, on each of the three fecal egg-counting devices. In addition, both manual (laboratory technician) and automated analysis (image analysis algorithm) were performed on the Telenostic device to calculate fecal egg counts (FEC). Overall, there were consistent egg counts reported across the three devices and calculation methods. The Telenostic device compared very favourably to the Mini-FLOTAC and McMaster. Only in sample C, a significant difference (P < 0.05) was observed between the egg counts obtained by Mini-FLOTAC and by the other methods. From this limited dataset it can be concluded that the Telenostic-automated test is comparable to currently used benchmark FEC methods, while improving the workflow, test turn-around time and not requiring trained laboratory personnel to operate or interpret the results.

Comparing the Mini-FLOTAC and centrifugal faecal flotation for the detection of coccidia (Eimeria spp.) in kiwi (Apteryx mantelli).

Coccidia (Eimeria spp.) in brown kiwi (Apteryx mantelli) cause significant morbidity and mortality in captive rearing facilities. Monitoring the abundance of this parasite in individual birds is crucial for successful management of kiwi. This research compares the abilities of centrifugal faecal flotations (CFF) and a modified Mini-FLOTAC protocol to detect oocysts. We hypothesised that the Mini-FLOTAC would detect higher oocyst counts. Kiwi dropping samples (n = 10) were homogenized in MgSO4 (SG 1.28) and oocyst counts made with CFFs and Mini-FLOTAC counting chambers, with three replicates for each method. For CFF, 0.5 g of droppings were examined using standard methods. Mini-FLOTAC counts were made using a modified sample preparation compared with the manufacturer's protocol but still used a 1:20 dilution of droppings. Oocysts were quantified using light microscopy at ×100-300 magnification. A linear mixed-effects model by REML showed that oocyst per gram estimates via the Mini-FLOTAC method were 3.2 times higher (95% CI 2.4-4.5, p < 0.01) than the CFF results. This increased detection likely represents a more accurate estimation of parasite shedding and should be considered for use in research or applications requiring more accuracy, cost-effectiveness, or accessibility than the CFF provides.

The recovery of added nematode eggs from horse and sheep faeces by three methods.

BACKGROUND: Nematode infections in horses are widespread across the world. Increasing levels of anthelmintic resistance, reported worldwide in equine parasites, have led to the creation of programs for the control of nematodes based on faecal egg counts (FEC). To improve nematode egg counting in equine faecal samples and establish whether the matrix of equine faeces or the eggs affect the counts, the analytical sensitivity, accuracy and precision of Mini-FLOTAC (combined with Fill-FLOTAC), McMaster and Cornell-Wisconsin techniques were compared. Known numbers of eggs extracted from equine or ovine faeces were added to egg free ovine and equine faeces to give counts of 10, 50, 200 and 500 eggs per gram (EPG) of faeces. RESULTS: The Cornell-Wisconsin significantly underestimated egg counts and McMaster showed a low analytical sensitivity, revealing 100% of sensitivity only for concentrations greater than 200 EPG. EPG values detected by Mini-FLOTAC did not differ significantly from expected counts at any level of egg density. CONCLUSIONS: Mini-FLOTAC combined to Fill-FLOTAC which provides an accurate method of weighing without need for a balance and filtering out debris, could be used for FEC on the farm as well as in the laboratory.

Mini-FLOTAC for counting Toxoplasma gondii oocysts from cat feces--comparison with cell counting plates.

Oocysts of Toxoplasma gondii represent one of the most common environmental contaminants causing the zoonotic infection toxoplasmosis. The aim of the present study was to compare the Mini-FLOTAC device with traditional cell counting plates (Kova Slide) for the detection of T. gondii oocysts from feline feces. Two types of experiments were performed: (i) purified oocysts were counted in different dilutions and (ii) specific pathogen free T. gondii-negative cat feces was inoculated with numbers of purified oocysts and counting was performed directly from feces. Our analysis showed a thousand times higher sensitivity of Mini-FLOTAC (5 × 10(2) oocysts) compared to Kova Slide (5 × 10(5) oocysts). Also, when compared by McNemar's test, counting of the purified oocysts showed a higher sensitivity of Mini-FLOTAC compared to Kova Slide, for a dilution of 10(3) oocysts/ml (chi(2) = 6.1; P < 0.05). A better sensitivity was also found with Mini-FLOTAC in dilutions of 10(5) and 10(4) oocysts/ml, when counted from feces (chi(2) = 4.2 and 8.1, respectively, P < 0.05). Our results show that Mini-FLOTAC is more sensitive than traditional methods of T. gondii oocysts detection and quantification is more accurate. Furthermore, Mini-FLOTAC simplicity and cost effectiveness allow it to be used with light microscopes in any laboratory or field conditions. We therefore recommend its use for regular screening. Further studies are needed to validate Mini-FLOTAC for the detection of oocysts in soil and water samples in field conditions.

Evaluation of Parasight All-in-One system for the automated enumeration of helminth ova in canine and feline feces.

BACKGROUND:  Digital imaging combined with deep-learning-based computational image analysis is a growing area in medical diagnostics, including parasitology, where a number of automated analytical devices have been developed and are available for use in clinical practice. METHODS: The performance of Parasight All-in-One (AIO), a second-generation device, was evaluated by comparing it to a well-accepted research method (mini-FLOTAC) and to another commercially available test (Imagyst). Fifty-nine canine and feline infected fecal specimens were quantitatively analyzed by all three methods. Since some samples were positive for more than one parasite, the dataset consisted of 48 specimens positive for Ancylostoma spp., 13 for Toxocara spp. and 23 for Trichuris spp. RESULTS: The magnitude of Parasight AIO counts correlated well with those of mini-FLOTAC but not with those of Imagyst. Parasight AIO counted approximately 3.5-fold more ova of Ancylostoma spp. and Trichuris spp. and 4.6-fold more ova of Toxocara spp. than the mini-FLOTAC, and counted 27.9-, 17.1- and 10.2-fold more of these same ova than Imagyst, respectively. These differences translated into differences between the test sensitivities at low egg count levels (< 50 eggs/g), with Parasight AIO > mini-FLOTAC > Imagyst. At higher egg counts Parasight AIO and mini-FLOTAC performed with comparable precision (which was significantly higher that than Imagyst), whereas at lower counts (> 30 eggs/g) Parasight was more precise than both mini-FLOTAC and Imagyst, while the latter two methods did not significantly differ from each other. CONCLUSIONS: In general, Parasight AIO analyses were both more precise and sensitive than mini-FLOTAC and Imagyst and quantitatively correlated well with mini-FLOTAC. While Parasight AIO produced lower raw counts in eggs-per-gram than mini-FLOTAC, these could be corrected using the data generated from these correlations.

Assessing the efficacy of the ovicidal fungus Mucor circinelloides in reducing coccidia parasitism in peacocks.

The biological control of gastrointestinal (GI) parasites using predatory fungi has been recently proposed as an accurate and sustainable approach in birds. The current study aimed to assess for the first time the efficacy of using the native ovicidal fungus Mucor circinelloides (FMV-FR1) in reducing coccidia parasitism in peacocks. For this purpose, an in vivo trial was designed in the resident peacock collection (n = 58 birds) of the São Jorge Castle, at Lisbon, Portugal. These animals presented an initial severe infection by coccidia of the genus Eimeria (20106 ± 8034 oocysts per gram of feces, OPG), and thus received commercial feed enriched with a M. circinelloides suspension (1.01 × 108 spores/kg feed), thrice-weekly. Fresh feces were collected every 15 days to calculate the coccidia shedding, using the Mini-FLOTAC technique. The same bird flock served simultaneously as control (t0 days) and test groups (t15-t90 days). The average Eimeria sp. shedding in peacocks decreased up to 92% following fungal administrations, with significant reduction efficacies of 78% (p = 0.004) and 92% (p = 0.012) after 45 and 60 days, respectively. Results from this study suggest that the administration of M. circinelloides spores to birds is an accurate solution to reduce their coccidia parasitism.

Smells Like Anthelmintic Resistance-Gastrointestinal Prevalence, Burden and Diversity in Dogs from Portugal.

Anthelmintic resistance has been documented worldwide, but few cases have been reported in dogs. Shelter dogs are a great opportunity to study intestinal helminths and assess their potential resistance to anthelmintic compounds. With these two goals in mind, 361 faecal samples were collected from dogs in 18 municipalities in Portugal, from 19 shelters and 11 private tutors. These samples were analysed using the Mini-FLOTAC before and 13 days after deworming. The percentage of faecal egg count reduction was calculated. Parasitic forms were identified in 22.4% of the samples collected: Trichuris vulpis (10.5%), Toxocara canis (8.0%), members of the family Ancylostomatidae (6.4%), Toxascaris leonina (0.8%), Dipylidium caninum (0.6%) and members of the family Taeniidae (0.3%). The first three nematode parasites showed the highest intensity of infection (2900, 1325, and 1265 eggs per gram, respectively). In the second faecal collection, parasites were present in 20.8% of the samples. The anthelmintics febendazole + pyrantel embonate + praziquantel and praziquantel + fenbendazole were ineffective for T. vulpis and members of the family Ancylostomatidae in 16.2% and 6.9% of the samples, respectively. The parasites identified are all potentially zoonotic. This study is the first to suggest a potential resistance of T. vulpis to anthelmintics.

Comparison of the Automated OvaCyte Telenostic Faecal Analyser versus the McMaster and Mini-FLOTAC Techniques in the Estimation of Helminth Faecal Egg Counts in Equine.

Gastrointestinal helminth parasites continue to be a significant threat to the health of equine. OvaCyte Telenostic (OCT) (Telenostic Ltd., Kilkenny, Ireland) has developed an automated digital microscope utilising Artificial Intelligence to identify and count the clinically important helminth species in equine, bovine, and ovine host species. In this paper, the performance of the OCT analyser was evaluated for the detection and counting of equine helminth species parasites and its performance compared to the currently accepted benchmark methods of faecal egg counts being the McMaster and the Mini-FLOTAC techniques. A pairwise comparison of tests was assessed based on the correlation of egg counts and Cohen's kappa agreement statistics for dichotomized outcomes. Bayesian latent class analysis was used to estimate the sensitivity and specificity of all three techniques in the absence of a gold standard for four helminth parasites (strongyles, Anoplocephala spp., Parascaris spp. and Strongyloides westeri). Based on the analysis of 783 equine faecal samples, we found a high level (ρ ≥ 0.94) of correlation between each pairwise comparison of techniques for strongyle egg counts. Cohen's kappa agreement between techniques was high for strongyles and S. westeri, moderate for Parascaris spp., and low for Anoplocephala spp. All three techniques had a high sensitivity and specificity (>0.90) for strongyles. Across helminth parasites, the sensitivity of the OCT was the highest of the three techniques evaluated for strongyles (0.98 v 0.96 and 0.94), Anoplocephala spp. (0.86 v 0.44 and 0.46) and Parascaris spp. (0.96 v 0.83 and 0.96); but lowest for S. westeri (0.74 v 0.88 and 0.88), compared to McMaster and Mini-FLOTAC, respectively. In terms of specificity, OCT was the lowest in two species (Parascaris spp. 0.96, Anoplocephala spp. 0.95). In conclusion, OCT has a sensitivity and specificity statistically similar to both McMaster and Mini-FLOTAC, and had a higher correlation with Mini-FLOTAC. The OCT point of care faecal analyser offers improved workflow, test turn-around time and does not require trained laboratory personnel to operate or interpret the results.

Comparison of Mini-FLOTAC, Flukefinder and sedimentation techniques for detection and quantification of Fasciola hepatica and Calicophoron daubneyi eggs using spiked and naturally infected bovine faecal samples.

BACKGROUND: Fasciolosis (Fasciola hepatica) and paramphistomosis (Calicophoron daubneyi) are two important infections of livestock. Calicophoron daubneyi is the predominant Paramphistomidae species in Europe, and its prevalence has increased in the last 10-15 years. In Italy, evidence suggests that the prevalence of F. hepatica in ruminants is low in the southern part, but C. daubneyi has been recently reported at high prevalence in the same area. Given the importance of reliable tools for liver and rumen fluke diagnosis in ruminants, this study evaluated the diagnostic performance of the Mini-FLOTAC (MF), Flukefinder(R) (FF) and sedimentation (SED) techniques to detect and quantify F. hepatica and C. daubneyi eggs using spiked and naturally infected cattle faecal samples. METHODS: Briefly, negative bovine faecal samples were artificially spiked with either F. hepatica or C. daubneyi eggs to achieve different egg count levels: 10, 50 and 100 eggs per gram (EPG) of faeces. Moreover, ten naturally infected cattle farms from southern Italy with either F. hepatica and/or C. daubneyi were selected. For each farm, the samples were analysed individually only with MF technique and as pools using MF, FF and SED techniques. Bayesian latent class analysis (LCA) was used to estimate sensitivity and accuracy of the predicted intensity of infection as well as the infection rate in the naturally infected farms. RESULTS: The outcome of this study showed that the highest number of eggs (F. hepatica and C. daubneyi) recovered was obtained with MF, followed by FF and SED in spiked infected samples at 50 and 100 EPG, while at lower infection levels of 10 EPG, FF gave the best results. Moreover, the sensitivity for all the techniques included in the study was estimated at > 90% at infection levels > 20 EPG for both F. hepatica and C. daubneyi eggs. However, MF was the most accurate of the three techniques evaluated to estimate fluke infection intensity. Nevertheless, all three techniques can potentially estimate infection rate at farm level accurately. CONCLUSIONS: Optimization and standardization of techniques are needed to improve the FEC of fluke eggs.

Evaluation of Fecal Egg Count Tests for Effective Control of Equine Intestinal Strongyles.

The American Association of Equine Practitioners strongly advocates evidence-based intestinal strongyle control in horses. It recommends targeted treatment of all heavy egg shedders (>500 eggs per gram (EPG) of feces), while the low shedders (0-200 EPG) are left untreated. As 50-75% of adult horses in a herd are low shedders, preventing them from unnecessary anthelmintic exposure is critical for tackling resistance. There are various fecal egg count (FEC) techniques with many modifications and variations in use, but none is identified as a gold standard. The hypothesis of the study was that the diagnostic performance of 12 commonly used quantitation methodologies (three techniques with four variants) differs. In this regard, method comparison studies were performed using polystyrene beads as proxy for intestinal strongyle eggs. Mini-FLOTAC-based variants had the lowest coefficient of variation (CV%) in bead recovery, whereas McMaster variants had the highest. All four variants of Mini-FLOTAC and the NaNO3 1.33 specific gravity variant of modified Wisconsin followed a linear fit with R2 > 0.95. In contrast, the bead standard replicates for modified McMaster variants dispersed from the regression curve, causing a lower R2. The Mini-FLOTAC method seems less influenced by the choice of floatation solution and has better repeatability parameters and linearity for bead standard recovery. For FEC tests with high R2 (>0.95) but that underestimated the true bead count, a correction factor (CF) was determined to estimate the true count. Finally, the validity of CF was analyzed for 5 tests with R2 > 0.95 to accurately quantify intestinal strongyle eggs from 40 different horses. Overall, this study identified FEC methodologies with the highest diagnostic performance. The limitations in standardizing routine FEC tests are highlighted, and the importance of equalization of FEC results is emphasized for promoting uniformity in the implementation of parasite control guidelines.

Sensitivity comparison between Mini-FLOTAC and conventional techniques for the detection of Echinococcus multilocularis eggs.

Canines serve as the definitive host of Echinococcus multilocularis. This study evaluated the sensitivity of the Mini-FLOTAC technique (MF) for the detection of E. multilocularis eggs in definitive hosts. First, we investigated the effects of heat inactivation and preservative conditions on the detection rate of eggs obtained from experimentally infected dogs. The sensitivity of MF was compared with that of eight other techniques: the centrifugal flotation with sucrose or zinc sulfate, MGL, AMS III, and a combination of MF and flotation/sedimentation techniques. Finally, we compared the sensitivity of MF and the centrifugal flotation with sucrose for the feces of E. multilocularis-infected foxes. The detection rate reached a plateau level with a specific gravity (s.g.) 1.22 for fresh eggs, but the highest rates were obtained with s.g. greater than 1.32 for heat-inactivated eggs. There was no significant difference in the detection rate among the preservative conditions. MF showed significantly higher EPG than the other techniques. Moreover, it showed higher diagnostic sensitivity for the fox feces than the centrifugal flotation technique. These results suggest that heat inactivation may alter s.g. of E. multilocularis eggs and that MF with zinc sulfate (s.g. = 1.32) would be effective for detecting heat-inactivated E. multilocularis eggs.

Anthelmintic resistance of horse strongyle nematodes to fenbendazole in Lithuania.

BACKGROUND: Control of strongyle infections presents a global challenge for horse practitioners due to the development of anthelmintic resistance (AR), however comprehensive information on AR in Lithuania is still lacking. The aim of this study was to assess the current situation of fenbendazole (FBZ) AR in horses at stable level in Lithuania. RESULTS: Faecal samples from 121 horses from six stables were examined using the Mini-FLOTAC method. Of these, 89 horses met the inclusion criteria that included strongyle faecal egg counts (FEC) exceeding 200 eggs per gram (EPG). Faecal egg count reduction tests (FECRTs) were performed in these. AR was evaluated at horse stable level based on faecal egg count reduction (FECR) and the lower limit of the 95% credible interval (LLCI) using the Bayesian hierarchical model. This study confirmed that strongylids (Cyathostominae (CYA)) resistant to FBZ are pervasive in Lithuania. FBZ was ineffective in three of the six stables (FECR 77.1-79.0%; 49.8-99.8 LLCI), was suspected to be ineffective in one stable (FECR 93.6%; 85.4-100 LLCI) and was effective (FECR 99.8-100%; 99.8-100 LLCI) in two stables. FEC showed a significant (P < 0.01) difference between the treatment and control groups. Only CYA larvae were detected in larval cultures derived from strongyle-positive faecal samples collected 14 days after treatment of a test group with FBZ. CONCLUSION: This in vivo study showed that resistance to FBZ in the treatment of strongyle nematodes is prevalent in horse stables in Lithuania. These findings should guide the implementation of more sustainable management of strongyle infections in horses in Lithuania.

A comparison of Mini-FLOTAC and McMaster techniques, overdispersion and prevalence of parasites in naturally infected North American bison (Bison bison) in the USA.

Several quantitative diagnostic techniques are available to estimate gastrointestinal parasite counts in the feces of ruminants. Comparing egg and oocyst magnitudes in naturally infected samples has been a recommended approach to rank fecal techniques. In this study, we compared the Mini-FLOTAC (sensitivity of 5 eggs per gram (EPG)/oocysts per gram (OPG)) and different averaged replicates of the modified McMaster techniques (sensitivity of 33.33 EPG/OPG) in 387 fecal samples from 10 herds of naturally infected North American bison in the Central Great Plains region of the USA. Both techniques were performed with fecal slurries homogenized in a fill-FLOTAC device. In the study population, prevalence of strongyle eggs, Eimeria spp. oocysts, Moniezia spp. eggs and Trichuris spp. eggs was 81.4%, 73.9%, 7.5%, and 3.1%, respectively. Counts of strongyle eggs and Eimeria spp. oocysts obtained from 1 to 3 averaged technical replicates of the modified McMaster technique were compared to a single replicate of the Mini-FLOTAC. Correlation between the two techniques increased with an increase in the number of averaged technical replicates of the modified McMaster technique used to calculate EGP/OPG. The correlation for Moniezia spp. EPG when averaged triplicates of the modified McMaster technique were compared to a single replicate of the Mini-FLOTAC count was high; however, the correlation for Trichuris spp. eggs was low. Additionally, we used averaged counts from both techniques to show the overdispersion of parasites in bison herds.