Mini-FLOTAC and McMaster egg counting method for detection of gastrointestinal parasites in small ruminants: a comparison study

The proper diagnosis of gastrointestinal parasites of small ruminants requires the development of multivalent techniques characterized by high sensitivity, specificity, precision, reproducibility, and the ability to quickly detect and monitor infections that pose risks to animal health. The aim of this study was to evaluate the performance of Mini-FLOTAC technique and McMaster egg counting (gold standard) in the detection of gastrointestinal parasites of goats and sheep. A total of 789 fecal samples were analyzed (401 of sheep and 388 of goats). From those, nematode eggs were detected in 80.86% (638/789), being 72.57% (291/401) from sheep and 89.43% (347/388) from goats. The Mini-FLOTAC technique presented a better performance to detect helminth eggs in both goats (88.40%; 343/388) and sheep (71.57%; 287/401) (?2 = 10.358; p < 0.0001). Kappa analysis revealed a weak concordance between techniques for goats (k = 0.342; p < 0.001) and sheep (k = 0.281; p < 0.001). Data herein reported suggests that the Mini-FLOTAC is a technique more sensitive than the McMaster egg counting, therefore its use might be adopted for the diagnosis of gastrointestinal infection in small ruminants.

Improving the sensitivity of gastrointestinal helminths detection using the Mini-FLOTAC technique in wild birds

High-performance validated tests are essential for successful epidemiological monitoring, surveillance of parasitic infections, and comparative studies in wildlife populations. The Mini-FLOTAC is a novel flotation-based technique for the sensitive detection and quantification of gastrointestinal parasites that is recently being explored for use in wildlife. A limitation of any flotation-based copromicroscopic method is the selection of the flotation solution (FS), which might influence the performance of the test. However, no study has compared the influence of using different FS in the Mini-FLOTAC technique for parasite detection in wild birds. Here, we evaluated the diagnostic performance of the Mini-FLOTAC in three waterbird host species using two widely used FS: saturated salt (NaCl; specific gravity 1.20) and saturated zinc sulfate (ZnSO4; specific gravity 1.35). One hundred fresh fecal samples were analyzed for parasite fecal egg counts (FEC). Regardless of the host species, fecal samples evaluated with the Mini-FLOTAC method using ZnSO4 resulted in a significantly higher detection rate and higher FEC of strongylid, capillarid, cestode, and trematode parasites, than samples analyzed with the NaCl solution. Our concise study demonstrated the importance of using an appropriate FS for the identification of parasite eggs in wildlife species, especially in hosts with an expected aggregated distribution and low parasite load such as waterbird hosts. The higher analytical sensitivity of the Mini-FLOTAC technique achieved with ZnSO4, and its applicability to fieldwork, highlights this method as a promising tool for the quantitative surveillance of parasite infections in wild bird populations.

Multispecific resistance of sheep trichostrongylids in Austria

Anthelmintic overuse and failure to implement methods preventing the development and spread of anthelmintic resistance (AR) have led to an alarming increase of resistant ovine trichostrongylids worldwide. The aim of the present study was to determine whether the routine anthelmintic treatment strategy was effective, to obtain insights into the frequency of AR in trichostrongylids of sheep in Austria, and to determine the presence of different trichostrongylid genera. On 30 sheep farms, the faecal egg count reduction test (FECRT) was performed with the Mini-FLOTAC technique in two consecutive studies. In study 1, only fenbendazole and moxidectin were tested, while different compounds and products were used in study 2. Overall, 33 treatment groups were formed: 11 groups were treated with benzimidazoles (fenbendazole and albendazole), 2 groups with avermectins (ivermectin, doramectin), 18 groups with moxidectin, and two groups with monepantel. Reduced efficacy was detected in 64%, 100%, 28% and 50% of these groups, respectively. The most frequently detected genus in larval cultures was Haemonchus, which had been barely detected in Austria previously, followed by Trichostrongylus. Multispecific resistance of trichostrongylids in Austria seems to be on the rise and H. contortus was detected unexpectedly frequently in comparison to previous studies. There is an urgent need to develop efficient communication strategies aimed at improving the engagement of farmers and veterinarians in sustainable parasite control.

Comparison of McMaster and Mini-FLOTAC effi ciency methods in diagnostics of Ascaris suum infection in young pigs

The coproovoscopic methods involving count chambers are widely used in the veterinary practice for diagnostics. These methods are important in assessments of the prevalence rates of nematode infections in pigs and for FECR tests. However, more basic data is needed on the efficiency of the commercial methods of diagnostics, tested at the different specific gravity of the flotation solutions. In a thorough analysis of publications of Ukrainian scientists, we have found that the data is sparse on the efficiency comparison of the modified McMaster’s and MiniFLOTAC methods. The aim of our study was to compare the efficiency of the commercial coproovoscopic techniques: the modified McMaster’s method (with sensitivity of 25 EPG), and MiniFLOTAC combined with Fill-FLOTAC (with sensitivity of 5 EPG) in testing for A. suum infection in piglets. Results of approbation of the mentioned methods are presented for the following saturated solutions: NaCl (SG=1.2), MgCl2 •6H2 O (SG =1.27) and NH4 NO3 (SG=1.28). Fecal samples were taken at own private farm, in Mlynok village of Onufriivka district, Kirovohrad region from 20 4 month old pigs with spontaneous A. suum infection. The laboratory assessments were performed in the Scientific Laboratory of Parasitology and Veterinary and Sanitary Expertise of the Poltava State Agrarian Academy. It is established that at high rates of A. suum infection in young pigs, McMaster’s method with the ammonium nitrate solution is more efficient compared to the Mini-FLOTAC method. In average, 5331.2 eggs of A. suum were determined per 1 g of feces using the former method, compared to 4567.5 EGP according to the Mini-FLOTAC technique (P < 0.001). The highest diagnostic efficiency of the modified McMasters method and the combination of Mini-FLOTAC and Fill-FLOTAC is seen when ammonium nitrate (NH4 NO3 ) solution of 1.28 SG is used (P < 0.001). Our results add new data on the efficiency of the commercial methods of diagnostics with varying specific gravity of the flotation solutions, for pigs infected with nematodes. Key words: EPG, pigs, nematode, ascariasis, diagnostics, efficiency.

PSVIII-11 Eimeria management for recently transported Holstein heifers using essential oils, anti IL-10 or monensin

Eimeria prevention in dairy heifers is challenging. Possible tools beyond the common practice of feeding an ionophore include polyphenolic compounds in essential oils (cinnamaldehyde, thymol, oregano oil) and binding interleukin-10 (IL-10) to reduce Eimeria’s host immunotolerance strategy. Holstein heifers (160 total; 12–14 wk old) were randomly assigned to one of 5 treatments: 160 mg/head/d of sodium monensin (MON), 1100 mg/head/d of antibody to IL-10 during wk 3–4 (aIL-10), 10 g/head/d of a blend of essential oils (EO), egg control without antibodies to IL-10 (EC), or negative control (NC). Calves were grouped 8 per pen with 4 blocks of 5 pens and FEC averaged by pen before analysis. We evaluated treatment effects on the fecal egg counts (FEC) of E. bovis, E. zurnii and E. auburnesis. Fecal counts were conducted biweekly using FLOTAC technique for 10 wk. Effects were analyzed with a mixed model ANOVA in SAS with repeated measures and ranked for normality. Total FEC prevalence was absent at wk 0, with greater prevalence at wk 4 and 6 for all treatments except MON (P &lt; 0.05), and there were no differences among treatments in positive prevalence by wk 8 and 10 (P &gt; 0.10). There was no difference between NC and MON treatments for wk 6, 8 and 10 in E. bovis, wk 2 and 4 for E. zurnii, and wk 2 and 4 for E. auburnesis (P &gt; 0.05), which is evidence of reduced monensin efficacy or excessive environmental Eimeria load. EO and aIL-10 FEC were also not different than MON counts at multiple timepoints (P &gt; 0.05). Overall, this shows potential monensin resistance after extended exposure (6–8 wk) to high environmental loads of Eimeria and similar FEC between monensin treated calves and those treated with EO or aIL-10.

Cattle gastrointestinal nematode egg-spiked faecal samples: high recovery rates using the Mini-FLOTAC technique

Background : Faecal egg count (FEC) techniques are commonly used to detect gastrointestinal nematode (GIN) in cattle and to determine anthelmintic efficacy/resistance through the faecal egg count reduction test (FECRT). Mini-FLOTAC is one of the techniques recommended for a standardised FEC/FECRT of helminth eggs in cattle. However, only one paper evaluated the recovery rate of GIN eggs by Mini-FLOTAC (compared to McMaster and modified-Wisconsin) in cattle, using only the level of contamination of 200 eggs per gram (EPG) of faeces and using GIN eggs collected from goat faeces to spike faecal samples from cattle. To further study the recovery rate of added GIN eggs from cattle, this study was conducted in two laboratories, one in Belgium and one in Italy to evaluate sensitivity, accuracy, precision and reproducibility of Mini-FLOTAC and McMaster (at two reading levels: grids and chambers) for the detection of GIN eggs in spiked bovine faecal samples. Methods: In both countries, spiked cattle faecal samples with five different levels of egg contamination (10, 50, 100, 200 and 500 EPG) of GINs were used. The study was performed in both laboratories by the same expert operator and using the same standard operating procedures (SOPs) for Mini-FLOTAC and McMaster. Sensitivity, accuracy and precision were calculated for each technique and for each level of contamination. Moreover, statistical analyses were performed to evaluate differences between performances of techniques used. Results : Mini-FLOTAC had a higher sensitivity (100% at all EPG levels for Mini-FLOTAC vs 0-66.6% for McMaster chambers and grids at levels <100 EPG), a higher accuracy (98.1% mean value for Mini-FLOTAC vs 83.2% for McMaster grids and 63.8% for McMaster chambers) and a lower coefficient of variation (10.0% for Mini-FLOTAC vs 47.5% for McMaster grids and 69.4% for McMaster chambers) than McMaster. There was no significant difference between the recovery of GIN eggs from the two studies performed in Belgium and in Italy. Conclusions : The high GIN eggs recovery rate detected by Mini-FLOTAC and the similar results obtained in Belgium and in Italy indicated that the diagnostic performance of a FEC technique was not regard to the laboratory environment.

Cattle gastrointestinal nematode egg-spiked faecal samples: high recovery rates using the Mini-FLOTAC technique

Background: Faecal egg count (FEC) techniques are commonly used to detect gastrointestinal nematode (GIN) in cattle and to determine anthelmintic efficacy/resistance through the faecal egg count reduction test (FECRT). Mini-FLOTAC is one of the techniques recommended for a standardised FEC/FECRT of helminth eggs in cattle. However, only one paper evaluated the recovery rate of GIN eggs by Mini-FLOTAC (compared to McMaster and modified-Wisconsin) in cattle, using only the level of contamination of 200 eggs per gram (EPG) of faeces and using GIN eggs collected from goat faeces to spike faecal samples from cattle. To further study the recovery rate of added GIN eggs from cattle, this study was conducted in two laboratories, one in Belgium and one in Italy to evaluate sensitivity, accuracy, precision and reproducibility of Mini-FLOTAC and McMaster (at two reading levels: grids and chambers) for the detection of GIN eggs in spiked bovine faecal samples. Methods: In both countries, spiked cattle faecal samples with five different levels of egg contamination (10, 50, 100, 200 and 500 EPG) of GINs were used. The study was performed in both laboratories by the same expert operator and using the same standard operating procedures (SOPs) for Mini-FLOTAC and McMaster. Sensitivity, accuracy and precision were calculated for each technique and for each level of contamination. Moreover, statistical analyses were performed to evaluate differences between performances of techniques used. Results: Mini-FLOTAC had a higher sensitivity (100% at all EPG levels for Mini-FLOTAC vs 0-66.6% for McMaster chambers and grids at levels <100 EPG), a higher accuracy (98.1% mean value for Mini-FLOTAC vs 83.2% for McMaster grids and 63.8% for McMaster chambers) and a lower coefficient of variation (10.0% for Mini-FLOTAC vs 47.5% for McMaster grids and 69.4% for McMaster chambers) than McMaster. There was no significant difference between the recovery of GIN eggs from the two studies performed in Belgium and in Italy. Conclusions: The high GIN eggs recovery rate detected by Mini-FLOTAC and the similar results obtained in Belgium and in Italy indicated that the diagnostic performance of a FEC technique was not regard to the laboratory environment.