Maria Elena Morgoglione

Monitoring drug efficacy against gastrointestinal nematodes when faecal egg counts are low: do the analytic sensitivity and the formula matter?

The faecal egg count reduction test (FECR) is the recommended technique to monitor anthelmintic drug efficacy in livestock. However, results are often inconclusive due to the low analytic sensitivity of the diagnostic technique or the conflict in results from FECR formulae. A novel experimental set-up was, therefore, used to compare the impact of analytic sensitivity and formulae on FECR results. Four McMaster techniques (analytic sensitivities 50, 33.3, 15 and 10) and a FLOTAC technique (analytic sensitivity ~ 1) were used on faecal samples of 30 calves with a FEC of less than 200 eggs per gram. True drug efficacies of 70%, 80% and 90% were experimentally mimicked by comparing FEC before and after dilution (3:10, 2:10 and 1:10, respectively). The FECR was summarized using group (FECR(1)) and individual (FECR(2)) based formulae. There was a significant increase in precision of FECR when the analytic sensitivity increased (p < 0.0001). The precision also depended on the formula used, FECR(1) (p < 0.05) resulting in more precise FECR compared to FECR(2). The accuracy of the FECR differed marginally between the two formulae (p = 0.06), FECR(1) being more accurate. In conclusion, the present study describes a novel methodology to compare techniques for the precision and the accuracy of their FECR results. The results underscored that techniques with high analytic sensitivity will improve the interpretation of FECR in animal populations where baseline FEC are low. They also point out that the precision of individual-based formulae is affected by the analytic sensitivity.

Comparison of faecal techniques including FLOTAC for copromicroscopic detection of first stage larvae of Angiostrongylus vasorum.

Angiostrongylus vasorum is a metastrongylid nematode that resides in the pulmonary arteries and the right heart chambers. In dogs, infection results in respiratory, bleeding and neurological disorders and further clinical signs. In the present study, FLOTAC was evaluated for the detection of first-stage larvae (L1) of A. vasorum in canine faecal samples. This technique is based on the counting of parasitic stages (eggs, larvae, oocysts and cysts) in chambers after spinning of faecal samples onto a surface. In a first step, nine flotation solutions were evaluated using faeces of two experimentally infected dogs. Zinc sulphate (specific gravity (s.g.) 1.2) and zinc sulphate plus potassium iodomercurate (s.g. 1.45) gave good results. However, with the latter technique, the larvae were slightly deformed. Subsequently, FLOTAC, using zinc sulphate, was compared through a randomisation technique with McMaster, flotation in tube and Baermann–Wetzel technique. The mean larvae per gramme (LPG) obtained by the FLOTAC for both dogs was significantly higher (P < 0.05) than those obtained by the other three techniques (the means of the other techniques all lie below the 95% CI of the mean LPG of the FLOTAC technique). In addition, the FLOTAC results were consistent across replicates with only Poisson (or random) variation between individual replicates. The other techniques appear to be less consistent with evidence of extra-Poisson variation in at least one of the two dogs across the replicates within each technique. The FLOTAC technique may contribute to an improvement of the ability to diagnose canine lungworm infections and represent a valuable alternative for larval counting of A. vasorum in faecal samples, especially following transport or storage where there may be limited larvae viability, and larval migration techniques cannot be used.

Ancylostoma caninum : calibration and comparison of diagnostic accuracy of flotation in tube, McMaster and FLOTAC in faecal samples of dogs

We performed a calibration of flotation in tube, McMaster and FLOTAC to determine the optimal flotation solution (FS) and the influence of faecal preservation for the diagnosis of Ancylostoma caninum in dogs, and compared the accuracy of the three copromicroscopic techniques. Among nine different FS, sodium chloride and sodium nitrate performed best for detection and quantification of A. caninum eggs. Faecal samples, either fresh or preserved in formalin 5%, resulted in higher A. caninum egg counts, compared to frozen samples or preserved in formalin 10% or sodium acetate-acetic acid-formalin. FLOTAC consistently resulted in higher A. caninum eggs per gram of faeces (EPG) and lower coefficient of variation (CV) than McMaster and flotation in tube. The best results in terms of mean faecal egg counts (highest value, i.e. 117.0EPG) and CV (lowest value, i.e. 4.8%) were obtained with FLOTAC using sodium chloride and faecal samples preserved in formalin 5%. Our findings suggest that the FLOTAC technique should be considered for the diagnosis of A. caninum in dogs.

Is gastrointestinal strongyle faecal egg count influenced by hour of sample collection and worm burden in goats

Gastrointestinal (GI) strongyle infections remain one of the main constraints to goat production worldwide. Although several indicators of GI strongyle infections have been proposed, faecal egg count (FEC) techniques are the most common approaches for the estimation of prevalence and intensity of these parasites. However, FEC may be subjected to a within-individual variation due to factors such as weather, season, random day-to-day variation and phase of the parasitic infection. A longitudinal study of GI strongyle FEC was conducted on 63 dairy goats to evaluate the effects of hour of sample collection on FEC and the relationship between FEC and worm burden. The goats were naturally infected with Teladorsagia circumcincta, Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum venulosum. Every 3 weeks for 14 months faeces were collected every 2 h for 24 h from three individually caged goats. The faeces were those passed by the goats during the 2 h preceding each collection. For each goat a sample from this material was analyzed using the Flotac double technique. To explore relationships between FECs and adult parasite counts, on the day following sampling, the three goats were euthanized and the nematodes present in the abomasa and intestines were recovered, identified and counted. The results of the general linear model did not show any significant effect of the hour of faecal sample collection on GI strongyle FEC, whereas a significant effect of the month of faecal sample collection was found. The results of the Pearson correlation showed a significant (P=0.000) positive relationship between FEC and worm burden (r=0.6), in particular regarding H. contortus (r=0.9). In conclusion, the present study showed that the hour of sample collection does not influence the GI strongyle FEC and that there is a good relationship between FEC and total GI strongyle worm burden in goats.

Gastrointestinal strongyle Faecal Egg Count in goats: circadian rhythm and relationship with worm burden

ABSTRACT Gastrointestinal strongyle Faecal Egg Count in goats:circadian rhythm and relationship with worm burden G. Cringoli & L. Rinaldi & V. Veneziano & S. Pennacchio & M. E. Morgoglione & M. Santaniello & M. Schioppi & V. Fedele # Springer Science + Business Media B.V. 2008 Keywords Faecaleggcount.Gastrointestinalstrongyles.GoatsIntroductionThe population of goats in the world has increased considerably over the past few years,also in Europe-in particular in the Mediterranean Basin-where 18 % of the world’s goatsmilk is produced, nearly exclusively for cheeses (Boyazoglu et al. 2005).The quality and quantity of dairy goat production, however, is negatively affected byseveral factors, including parasitic diseases. Among these, gastrointestinal (GI) strongyleinfection remains one of the main constraints to goat production throughout the world.The contemporaneous presence of different genera/species of these nematodes oftencauses inflammatory/traumatic problems, as well as a diversion of nutrients from productionsites towards the repair of tissue-damage provoked by the parasites, with negativerepercussions on the growth, fecundity and productivity of the animals (Hoste et al. 2005).For the above reasons, the diagnosis of these parasitic infections is of importance, withthe final goal of a targeted treatment.Although some physiopathological indicators of GI strongyle infections (e.g. anaemiaindex, diarrhoea index and body condition score) have been proposed in recent years,Faecal Egg Count (FEC) techniques remain the most common approach for the diagnosis of

Helminths and arthropoda in buffalo farms from the Lazio region (Italy)

ABSTRACT Helminths and arthropoda in buffalo farmsfrom the Lazio region (Italy) G. Cringoli & V. Musella & M. P. Maurelli & M. E. Morgoglione & A. Santaniello & R. Condoleo & I. Guariglia & L. Rinaldi Published online: 30 June 2009 # Springer Science + Business Media B.V. 2009 Keywords Arthropoda.Buffaloes.Helminths.LazioregionIntroductionInformation on the presence and distribution of helminths and arthropoda in water buffaloesfrom the Lazio region of Central Italy (where 18% of Italian water buffaloes are bred) isvery fragmentary and scant. In addition, they derive from studies conducted in past periodswhen buffaloes were mainly bred on pasture and when anthelmintic treatments were notwidely used (e.g. Rivellini et al., 1972). For these reasons, the present survey was aimed atobtaining up-to-date information on the distribution of helminths and arthropoda in waterbuffaloes from the Lazio region, where the farm management system is now characterizedby intensive breeding techniques.Materials and methodsThe survey was carried out on a sample of 127 buffalo farms in 20 contiguousmunicipalities located in the provinces of Latina and Frosinone. This sample size wascalculated using the formula proposed by Thrusfield (1995) inserting the following values:study population (531 buffalo farms), expected prevalence (12%), desired absoluteprecision (5%) and confidence level (95%). First of all, all the buffalo farms located inthe study area were geo-referenced within a Geographical Information System (GIS). The127 farms that comprised the sample were selected using a grid approach (Rinaldi et al.,

Helminths in Sheep on Farms of the Basilicata Region of Southern Italy

The aim of this study was to obtain up-to-date information on the presence and distribution of helminths in grazing sheep from the Basilicata region of southern Italy. A cross-sectional coprological survey was conducted on 98 sheep farms and the FLOTAC dual technique was used to detect and count parasitic elements. The most frequent nematodes found were gastrointestinal (GI) strongyles (91.8 %), in particular Haemonchus (76.9 %), Trichostrongylus (91.8 %), Teladorsagia (88.8 %), Cooperia (77.6 %), and Oesophagostomum (72.5 %). They were followed by lungworms (50.0 %), including Muellerius (37.8 %), Neostrongylus (13.3 %), Protostrongylus (4.4 %), Cystocaulus (2 %), and Dictyocaulus (2 %). We also found Trichuris (39.8 %), Nematodirus (24.5 %), Strongyloides (4.1 %), and Skrjabinema (2.0 %). Trematoda were also found with the following prevalence: Calicophoron daubneyi (10.2 %), Dicrocoelium dendriticum (61.2 %), and Fasciola hepatica (1.0 %). Moniezia was found on 35.7 % of farms. Interestingly, the spatial analysis showed clusters for C. daubneyi and D. dendriticum.

Ixodidae ticks in sheep and cattle in the Basilicata region (southern Italy)

Ixodidae are an important health problem for domestic and wild animals for direct damages caused by these ticks, but mostly because they are involved in the transmission of many diseases (tick borne diseases - TBDs). Therefore, the surveys on the presence and geographical distribution of ticks in domestic ruminants need to be constantly updated. The aim of the present study was to update the data on the presence and distribution of Ixodidae ticks in cattle and sheep bred in the Basilicata region (southern Italy), an area with a Mediterranean climate favourable to the biology and ecology of ticks. In this region, the breeding of pasturing cattle and sheep is a widespread reality very important from an economical point of view, especially for the natural vocation of animals in using marginal hilly and mountainous pastures. From May to September 2013, sheep and cattle farms (n = 82 and 31, respectively) were visited and ticks were collected from 20 animals per farm, preserved in ethanol 70% and then identified at species level using the morphometric keys present in the literature. A total of 2,179 ticks were collected. The following species were identified in the sheep farms (total = 1594 adult ticks collected): Rhipicephalus bursa (67.1%), R. sanguineus (34.1%), R. turanicus (25.6%), Dermacentor marginatus (6.1%), Hyalomma detritum (3.7%), Ixodes ricinus (3.7%) and Hyalomma marginatum (1.2%). In the cattle farms, a total of 585 adult ticks were collected and the following species identified: R. bursa (61.3%), H.marginatum (29.0%), R. turanicus (25.8%) sanguineus (25.8%), D. marginatus (9.7%), I. ricinus (9.7%), Ixodes gibbosus (6.5%) and Hyalomma detritum (3.2%). The results of the present cross-sectional survey confirm that the environmental and climatic conditions of southern Italy are suitable for different tick species infecting domestic ruminants that are therefore exposed to different TBDs.

A national survey of Ixodidae ticks on privately owned dogs in Italy

BackgroundThe geographical distribution of ticks on companion animals needs to be monitored to develop and plan effective control measures, as suggested by the European Scientific Counsel on Companion Animal Parasites. The aim of this study was to conduct the first Italian national survey of tick distribution on privately owned dogs.MethodsThe study was performed over 20 months (February 2016 - September 2017) and involved 153 veterinary practices in 64 different provinces covering 17/20 (85%) Italian regions. Participating practitioners were asked to examine five different dogs per month at random and complete a questionnaire for each dog. Differences in tick infestation associated with: sex, age and hair length (long and short); the dog’s habitat (indoor or outdoor/kennel); and the dog’s environment (urban or rural/sylvatic) were evaluated. The attachment site of ticks on the dog was also recorded. Acaricide efficacy was evaluated for the subset of dogs for which complete information on product used, date of sampling and date of last ectoparasiticide treatment was available.ResultsOf the 3026 dogs examined, 1383 (45.7%) were carrying at least one tick. Overall, 2439 tick samples were collected and a total of 14 tick species identified. Rhipicephalus sanguineus group were the most predominant ticks (63.6%), followed by Ixodes ricinus (30.6%) and I. hexagonus (5.6%). Twenty-four dogs had mixed tick infestations. Long-haired dogs had a higher tick infestation risk as did dogs with outdoor and rural/sylvatic lifestyles. Ticks were located on the head (37.4%), the neck (28.8%), the muzzle (15.5%) and the back (15.3%). A higher prevalence of Rhipicephalus was found in the interdigital spaces (10.8%) compared to Ixodes (0.2%). Finally, ectoparasiticide treatments were found significantly protective against tick infestation, especially orally administered formulations.ConclusionsPrivately owned dogs in Italy have a high prevalence (45.7%) of infestation with ixodid ticks and this risk varies by dog phenotype and lifestyle.

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

BackgroundNematode 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.ResultsThe 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.ConclusionsMini-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.