Paola Beraldo

Anthelmintic resistance in cyathostomin populations from horse yards in Italy, United Kingdom and Germany.

BackgroundA large survey was carried out in 2008 in Europe to evaluate the efficacy of fenbendazole (FBZ), pyrantel (PYR), ivermectin (IVM) and moxidectin (MOX), i.e. the major anthelmintic molecules used in current practice against cyathostomins affecting horses. A total of 102 yards and 1704 horses was studied in three countries: 60 yards and 988 horses from Italy, 22 and 396 from the UK, 20 and 320 from Germany. The survey consisted of Faecal Egg Count Reduction Tests (FECRTs) with a faecal egg count reduction (FECR) categorization of (I) resistance present if FECR <90% and the lower 95% confidence limit (LCL) <90%, (II) resistance suspected if FECR ≥ 90% and/or LCL <90% and (III) no resistance if FECR ≥ 90% and LCL >90%. The calculation of FECR data was performed employing bootstrap analysis of group arithmetic means.ResultsThe testing of FBZ on a total of 80 yards resulted in resistance present on more than 80% of the UK and German yards and on significantly fewer in Italy, i.e. in 38% (p < 0.01). PYR, IVM and MOX were tested on a total of 102 yards. For PYR resistance present was found in 25% of the yards with no significant differences between countries. For IVM resistance present was encountered in one Italian and two UK yards (3%), resistance present to MOX was not found in any yard in any country.ConclusionThe results indicate that single and/or multiple drug resistance in equine cyathostomins is present in the three countries, is widespread particularly for FBZ and/or PYR and in one UK yard multiple resistance present was detected to FBZ, PYR and IVM. Macrocylic lactones proved to be the most effective drugs, with some evidence of resistance to IVM and highest activity of MOX, despite a single case of reduced efficacy in Germany. These data call for the development and implementation, among practitioners, owners and managers, of further plans to reduce the expansion of the anthelmintic resistant populations and to use those anthelmintics that remain effective in a manner that preserves their efficacy as long as possible.

Growth response of juvenile dentex (Dentex dentex L.) to varying protein level and protein to lipid ratio in practical diets

Abstract The dentex ( Dentex dentex L.) is a fast-growing sparid which represents a possible candidate for Mediterranean mariculture. As the basic nutrient requirements of this species are unknown, two feeding trials were carried out to identify suitable protein and lipid (energy) levels to be used in formulating practical diets for dentex. The experiments used groups of 30 specimens (each fish weighing 17 or 20.7 g) which were kept in 12 flow-through 160 1 tanks supplied with sea water at 2 1 min −1 (temperature 20 ± 1 °C; salinity 33 ppt). An initial 4-week trial was conducted to obtain a preliminary estimate of the optimal protein level. Triplicate groups were fed four practical diets with increasing levels of protein (44.3, 49.3, 55.7 and 58.9% dry matter (DM), over 90% of which was supplied by white fish meal) and a constant lipid content (17.3% DM; mostly supplied by fish lipids) to satiation. An analysis of the nutrient-weight gain relationship indicated that the optimal dietary protein level was 49.3% DM. In a second 60 day experiment, duplicate groups of fish were fed six diets with three protein levels (44.4, 49.5 and 55.8% DM) and two lipid contents (12.0 and 17.3% DM) within each protein level, nearly to satiation. Growth performance was lowest in fish given diets containing 44.4 or 49.5% protein and 12% lipid and was improved ( P −1 calculated available energy) could be assumed as suitable levels for formulating practical diets for this species, providing high quality fish meal and oil are used.

Effect of ultrasound treatments of tomato pulp on microstructure and lycopene in vitro bioaccessibility

The influence of ultrasound treatments of tomato pulp on microstructure and lycopene in vitro bioaccessibility was investigated. To this purpose, samples were subjected to ultrasound at a frequency and amplitude of 24 kHz and 100 μm, respectively, for increasing lengths of time. Results showed that ultrasound was responsible for loss of tomato cell integrity, as well as a decrease in the degree of pectin esterification. In contrast, rheological measurements showed that ultrasonically treated tomato pulp had greater gel-like properties than an untreated sample. It was inferred that ultrasound promoted the formation of a new network due to hydrogen bonding and hydrophobic interactions among the de-esterified pectin molecules. Such a reinforcement of the tomato pulp structure resulted in a decrease in lycopene in vitro bioaccessibility of the ultrasonically treated tomato pulp, probably due to the fact that the presence of a stronger network may make lycopene less available to the digestion process.

Effects of worm control practices examined by a combined faecal egg count and questionnaire survey on horse farms in Germany, Italy and the UK.

BackgroundFor the control of worm infections, the strategic use of anthelmintics, often accompanied by additional farm and/or pasture management procedures, is currently applied on most horse farms in industrialized countries. However, the particular effects of the specific worm control procedures are often unclear and have only been investigated to a limited extent. We examined faecal egg count (FEC), faecal egg count reduction (FECR) and questionnaire data on farm and pasture management procedures. The aim of this study was to determine whether specific worm control practices reported to be applied in European horse farms affect worm prevalence.ResultsThis study involved 20 German, 26 Italian and 16 UK horse farms for each of which FEC were performed on a minimum of 16 horses. In total, 2029 horse faecal samples were quantitatively analysed for helminth eggs, resulting in 56.3% of the faecal samples being positive for strongylid eggs. The prevalence in the 742 German horse samples (48.1%) was significantly lower than that in the 737 Italian (61.1%) and the 550 UK (60.9%) samples. As expected, a significant effect of horse age on the infection prevalence was observed, with adult horses showing lower prevalences and lower mean FEC than foals and yearlings. The majority of the participating farms were stud farms (n = 29), followed by riding stables (n = 27) and racehorse stables (n = 6). The prevalence of strongyle infection by farm type differed between countries. While in Germany, horses on riding farms were significantly less often strongyle positive, in the UK horses on stud farms showed the lowest strongyle prevalences, whereas in Italy, no significant difference between farm types were seen. On all farms, horses received routine/preventive anthelmintic treatment. An effect of treatment frequency on strongyle prevalence was only encountered with adult horses. On farms performing more than one annual treatment, faecal samples were significantly less often positive. Furthermore, by comparing the FECR results of individual horses with their pre-treatment FEC, it was found that high pre-treatment FEC were associated with a significantly higher probability for a FECR below 90%.ConclusionOverall, age-dependent strongyle infection patterns and general worm control approaches were found to be similar on horse farms in the three countries. Also, a negative association of pre-treatment FEC and treatment efficacy was consistently found in all countries. However, mean strongyle prevalences and frequencies of anthelmintic treatments were considerably different. In addition to the age-dependent prevalence patterns, the finding of a possible negative association between high FEC and reduced FECR might argue for a focus on horses showing high pre-treatment FEC when monitoring anthelmintic treatment efficacy in the field.

Abnormalities of the operculum in gilthead sea bream (Sparus aurata): morphological description

The authors studied a population of 582 gilthead sea breams (Sparus aurata), aged 25–600 days, in order to describe the abnormalities of the opercular complex. Particular attention was paid to: (1) the different degrees of abnormality of the opercular complex as a whole and of the individual dermal bones (preopercle, interopercle, subopercle and opercle) that compose it; (2) the ways in which the operculum folds into the gill cavity and the relevant histological characteristics. This study discusses the origin and nature of the abnormalities. Though these abnormalities have a major impact on market value and product image, and are often linked to other disorders in fish, they are still difficult to eliminate. Gaining a basic understanding of these abnormalities will be a first step towards reducing their occurrence. D 2003 Elsevier Science B.V. All rights reserved.

Distribution and species-specific occurrence of cyathostomins (Nematoda, Strongylida) in naturally infected horses from Italy, United Kingdom and Germany

A broad scale study was carried out in 2008 to evaluate the distribution and species-specific occurrence of cyathostomin populations in horse yards from Europe. In total 102 properties and 3123 horses were included in Italy (60 yards and 1646 animals), United Kingdom (22 yards and 737 animals) and Germany (20 yards and 740 animals). Individual faecal samples were examined with a McMaster technique while pooled samples were subjected to the microscopic examination of in vitro cultured larvae and to a Reverse Line Blot (RLB) assay able to molecularly identify the most diffused 13 species of cyathostomins. All yards were positive for the presence of cyathostomins both at the McMaster technique and at the microscopic examination of cultured larvae. One thousand and nine hundred thirty-one horses (61.8%) showed a positive faecal egg count, i.e. 1110 (67.4%), 463 (62.8%) and 358 (48.3%) from Italy, UK and Germany respectively. Out of the 1931 positive animals 1133 (36.3%) showed a faecal egg count per gram >150, specifically 694 (42.2%) from Italy, 237 (32.2%) from UK and 202 (27.3%) from Germany. The molecular results showed that all 13 species that can be detected by the RLB were found in each of the three countries, with a range of 3-13 species present in individual yards. The five most prevalent were Cylicocyclus nassatus, Cylicostephanus longibursatus, Cyathostomum catinatum, Cylicocyclus goldi and Cyathostomum pateratum. The relevance of these results and related biological and epidemiological features are discussed, together with their significance for both future studies of cyathostomins and further intervention programs aiming to control the spread of anthelmintic-resistant populations.

Decreased strongyle egg re-appearance period after treatment with ivermectin and moxidectin in horses in Belgium, Italy and The Netherlands

The objective of the present study was to evaluate the efficacy of an oral treatment with ivermectin (IVM) or moxidectin (MOX) against gastro-intestinal strongyles in naturally infected horses by performing a faecal egg count reduction test (FECRT) and by monitoring the egg reappearance period (ERP) after treatment. Therefore, a field efficacy study with a randomised complete block design for each study site was conducted, with the individual animal as the experimental unit. At least 10 study sites in Italy, Belgium and The Netherlands were selected and animals were allocated to one of the two treatment groups based on the pre-treatment faecal egg counts (FEC). Animals were treated on Day 0 with an oral paste containing either IVM (at 0.2mg/kg bodyweight) or MOX (at 0.4 mg/kg bodyweight). After treatment, faecal samples were collected at least every fortnight during 56 days after treatment with IVM and during 84 days after MOX treatment. In total, 320 horses on 32 farms were examined. The FECRT on Day 14 indicated a 100% efficacy in 59 of the 64 treatment groups and >92% efficacy in the remaining 5 groups. The ERP was decreased for at least one of the anthelmintics on 17 out of 32 study sites (15 sites or 47% for MOX and 17 sites or 53% for IVM) and on 9 sites (28%) the ERP was decreased for both anthelmintics. On some of these study sites the efficacy declined at the end of the expected ERP, often with good efficacy 2 weeks earlier. Nevertheless, on 1, 3 and 5 study sites in Italy, Belgium and The Netherlands respectively, an efficacy below 90% for IVM and MOX was identified as soon as Day 42 or Day 56. In The Netherlands, the efficacy of IVM was below 90% from Day 28 or Day 35 after treatment on 1 site each. The present study reports a high efficacy of MOX and IVM in a FECRT 14 days after treatment, yet does indicate a shortened ERP for these treatments in more than half of the selected study sites.

Faecal Cyathostomin Egg Count distribution and efficacy of anthelmintics against cyathostomins in Italy: a matter of geography?

BackgroundIn the framework of a trial carried out in 2008 in Europe to evaluate the efficacy of major parasiticides against horse cyathostomins, pre- and/or post-treatment Faecal Egg Counts (FEC) were evaluated in a total of 84 yards and 2105 horses from nine different regions from the South, the Center, the North-Center and North-East of Italy. Specifically, on the basis of FECs of the horses present in each property, 60 out of the 84 yards were enrolled for a Faecal Egg Count Reduction Test (FECRT) using fenbendazole, pyrantel, ivermectin and moxidectin.ResultsOf the 1646 horses bred in the 60 recruited yards, 416 animals had a FEC between 50 and 150 Eggs Per Gram (EPG) of faeces and 694 a FEC >150 EPG (i.e. with total of 1110 positive animals). Of the 1110 positive animals, those with the highest FECs (i.e. 988) were included in the FECRT. The FECRT for four anthelmintic compounds showed remarkable differences in terms of prevalence of reduced and equivocal efficacy against cyathostomins in the different areas of Italy. Administration of fenbendazole and pyrantel resulted in resistance present or suspected in about half of the yards examined while resistance to ivermectin was found in one yard from central Italy and suspected resistance was detected in three more yards, one in each the North, the Center and the South. Treatment with moxidectin was 100% effective in all yards examined.ConclusionCyathostomin populations in the South and the Center of Italy were more susceptible to fenbendazole and pyrantel than the populations present in the Center-North and North-Eastern areas of Italy. Fenbendazole and/or pyrantel were ineffective in almost all properties from the North of Italy. The reasons for such a difference among the Italian regions in terms of FECs and efficacy of antiparasitic drugs are discussed, together with the role that veterinarians, and horse owners and managers should have for effective worm control programs in this country.

First detection of Cytauxzoon spp. infection in European wildcats (Felis silvestris silvestris) of Italy

Cytauxzoonosis is an emerging, tick-transmitted, protozoan disease affecting domestic and wild felids and caused by Cytauxzoon felis, Cytauxzoon manul and Cytauxzoon spp. This study aimed to determine the presence of infection with Cytauxzoon spp. in Felis silvestris silvestris in Italy, in order to enhance the comprehension of its pattern distribution among domestic cat populations. In addition, wildcats were tested for other endemic vector-borne pathogens in Italy. The carcasses of 21 F. s. silvestris were collected from central and northern regions of Italy. All the animals were submitted to necropsy and samples of the spleens were collected. Cytauxzoon infection was surveyed by a conventional PCR amplifying a portion of the SSU-rDNA of species of Piroplasmida. The samples were also screened for Anaplasma spp., Ehrlichia spp., Rickettsia spp., Babesia spp., Theileria spp., and Leishmania spp. using SYBR Green Real-Time PCR (rPCR) assays. Four animals (19%) were positive for Piroplasmida-PCR assay and three sequenced amplicons were obtained (14.3%), clustering with the Italian, Spanish, French and Romanian Cytauxzoon spp. isolates and with C. manul found in Mongolia. The samples were negative for the other pathogens screened. The present results showed that Cytauxzoon spp. may infect both F. s. silvestris and F. s. catus.

Evaluation of the physicochemical properties of gluten‐free pasta enriched with resistant starch

BACKGROUND The objective was to examine the potential use of resistant starch (RS) as a fibre-enriching ingredient in gluten-free pasta. Pasta was enriched with commercial RS type II (Hi-Maize™ 260) at 100-200 g kg-1 substitution of rice flour. The effects on the rheological properties of dough and pasta quality as a result of RS addition and the loss in RS due to the process were evaluated. RESULTS Dough water absorption was not influenced by the addition of RS. The cooking loss (CL) of RS-enriched samples was 30% lower than reference without fibre. The addition of RS significantly increased firmness of cooked pasta, and above 100 g kg-1 RS level of substitution samples showed a lower stickiness value. Dynamic rheological tests on pasta dough showed a higher storage modulus for fibre samples, indicating a higher number of elastically physical interactions. Loss in RS in uncooked pasta was about 31% compared with the initial amount added to the product. CONCLUSIONS The addition of RS improved the quality of gluten-free pasta owing to its ability to increase the firmness and decrease the CL and stickiness of cooked pasta. The product enriched with 200 g kg-1 RS can be considered a source of DF.