Poultry roundworms have re-emerged in laying hens in many European countries due to the increase in noncaged housing. This is worrying because, at high parasite loads, Ascaridia galli can impact birds' welfare, health, and productivity. Worm control is therefore an important aspect of the successful management of the egg production industry. In 2009, the Swedish Egg Association initiated a voluntary control program to tackle the problem and reduce the appearance of worms in table eggs by encouraging producers to submit fecal samples for analysis. Since the start of the program, its data have never been thoroughly explored. Moreover, after more than a decade of challenges, our understanding of how egg producers perceive worm infection is still inadequate. This study was therefore designed to address these issues. The research data in the present study are drawn from 2 sources. First, through the control program and second, through an online survey. We have summarized the control program's achievements and discussed its findings and limitations. Although this work contributes to existing knowledge of roundworm control in laying hens in general, it also identifies gaps in knowledge. In conclusion, the control program can be improved by incorporating more strategic sampling and utilizing well-suited diagnostic tools for better assessment of infection status. It is equally important to educate producers on anthelmintics (AH) use and the development of resistance.

Since the EU ban on battery cages, many studies have listed Ascaridia galli and Heterakis gallinarum as the most common roundworms in the European laying hen population. A complicating factor is that the eggs of these parasites are almost identical. Thus, lack of molecular diagnostic approaches has driven epidemiological studies to take on necropsy for species discrimination, which is labor and cost intensive. Here, we describe a novel diagnostic tool based on droplet digital PCR for simultaneous identification and absolute quantification of the eggs of both of these ascarids in chickens’ droppings using two different genus-specific primer-probe sets targeting the second internal transcribed spacer region (ITS-2) in the nuclear ribosomal (rRNA) gene array. No cross-reaction was observed when different combinations of DNA and species-specific primers and probes were tested. The lowest obtained frequency threshold for the detection of H. gallinarum in the presence of a constant A. galli DNA concentration was determined to be 0.8 %. After validation, we used the assay to analyze field samples collected from several Swedish laying hen farms. Out of 134 samples, 86 (64 %) were positive for A. galli while 11 (8.3 %) samples were positive for H. gallinarum. These samples were initially analyzed with flotation technique for detection of ascarid eggs. The results of the Cohen’s kappa indicated substantial agreement (85.8 %) between the two tests. In conclusion, we have validated a novel molecular-based diagnostic tool for quantification and differentiation between intestinal parasites of major importance in chickens with high precision. Although this study focuses on identification of parasites of laying hens, the findings may well have a bearing on all types of chicken production systems. The present study lays the groundwork for future research into epidemiology of these two important chicken parasite species.

In the 3 years since the first report of canine alveolar echinococcosis (AE) in Ontario, three additional cases have been diagnosed in the province. Of the four cases reported to date, three have had no known history of travel outside the province. It is possible that this development is an indication of previously unrecognized environmental contamination with Echinococcus multilocularis eggs in some areas of the province. If so, there is the potential for an emerging threat to human health. This article describes a local public health department's investigation of the possible exposure to E. multilocularis of a number of individuals who had had contact with the latest of the four cases of canine AE, and summarizes a comprehensive decision process that can be used by public health departments to assist in the follow-up of such exposures.

A case-control study was performed to investigate the association between colic of all types in Swedish horses and infection with the equine tapeworm Anoplocephala perfoliata. Colic cases were defined by clinical signs consistent with the presence of abdominal pain, and the control horses had no signs of colic within the last year but attended a clinic for other reasons. Blood and fecal samples were collected by veterinarian from 67 horses with signs of colic and 67 control horses. The sera were analyzed using serodiagnostic assay anti-12/13 kDa IgG(T) ELISA. The fecal samples, 30 g from each horse, were analyzed with a modified sugar salt flotation method with a density of 1.280. A significant association was found between the presence of A. perfoliata eggs in feces and colic with a 16 times higher risk of colic if eggs had been observed in fecal samples. However, there was no significant association between colic and the median OD-values in the serological diagnosis, nor when recommended cut-offs were used. The study concludes that A. perfoliata is a risk factor for colic in Swedish horses and it suggests that the modified flotation method can be used as a diagnostic tool for identifying horses at risk.

Echinococcus multilocularis is a parasite that can cause alveolar echinococcosis disease. After the first positive finding of E. multilocularis in Sweden in 2011, a consulting group with representatives from relevant authorities was summoned. In this group, all relevant information was shared, strategies for information dissemination and any actions to be taken due to the finding of E. multilocularis were discussed and decided. The present paper describes the actions taken during 2011 and the results thereof, including surveillance in animals, risk assessment for humans to become infected and recommendations given to the public. Further discussion about whether the parasite was introduced, and if so, how, as well as possible future development of the infection in animals and humans in Sweden and future actions are included.

parasite and usage of targeted selective therapy regimens basing treatment decisions on individual fecal egg counts from all horses on the farm. The established diagnostic method for S. vulgaris involves larval culture and subsequent morphological identification of third stage larvae under the microscope. Recently, a real-time PCR assay was developed and validated for the detection and semi-quantification of S. vulgaris eggs in equine fecal samples. The purposes of the present study were a) to determine the presence of S. vulgaris by real-time PCR in Danish and American horses on farms using vastly different anthelmintic treatment regimens and b) to evaluate the association between larval culture results and the PCR. A total of 991 horses representing 53 different horse farms in Denmark and Central Kentucky were studied. Fresh fecal samples were collected from all horses, and strongyle eggs retrieved for DNA extraction and subsequent real-time PCR analysis. Individual larval cultures were performed on the Danish part of the data set (662 horses on 42 farms). On the Danish farms, the S. vulgaris PCR prevalencewas found to be 9.2% on farms not basing parasite control on fecal egg counts, and 14.1% on farms using selective therapy. None of the 328 horses evaluated on 11 farms in Central Kentucky were PCR positive. Kappa-values indicated a moderate agreement between PCR and larval culture results, while a McNemar’s test revealed no statistical difference between the paired proportions. Statistically significant associations were found between PCR cycle of threshold (Ct) value groups and larval culture counts. Results indicate that both diagnostic methods can be useful for determining the occurrence of S. vulgaris in horses on farms, but that they both are affected by potential sources of error. Agreement between tests was moderate and no direct linear association could be found. The PCR results confirmed previous findings suggesting that S. vulgaris is highly dependent on the anthelmintic treatment intensity, and that selective therapy regimens can be associated with higher occurrence of this parasite.

Surveillance for the fox tapeworm, Echinococcus multilocularis, has been carried out in Sweden since 2000, with about 300 red foxes analysed annually. We report the first finding of E. multilocularis in Sweden, in a fox shot in December 2010 in the south-west of the country. A second infected fox shot in the same location was detected in March 2011. This paper describes the national monitoring programme and the ongoing work to estimate the prevalence and spread of the infection.

The objective of the study was to investigate different aspects on the efficacy of three anthelmintics on cyathostomin nematodes of Swedish horses. A faecal egg count reduction (FECR) test was performed on 26 farms. Horses were treated orally with recommended doses of ivermectin, pyrantel pamoate or fenbendazole. Faecal samples were collected on the day of deworming and 7, 14 and 21 days later. No resistance was shown against ivermectin; the FECR was constantly >99%. The effect of pyrantel was assessed as equivocal in 6 farms 14 days after treatment; the mean FECR was 99%. As many as 72% of the fenbendazole-treated groups met the criteria for resistance; the mean FECR was 86%, ranging from 56% to 100%. A re-investigation of two farms where pyrantel resistance had been suspected clearly revealed unsatisfactory efficacy of pyrantel on one of these farms; the FECR varied from 72% to 89%. Twenty-six of the horses previously dosed with pyrantel or fenbendazole, and which still excreted ≥150 eggs per gram of faeces 14 days after treatment, were dewormed with ivermectin and fenbendazole or pyrantel in order to eliminate the remaining cyathostomins. A total of 13 cyathostomin species were identified from horses that initially received fenbendazole and seven species were identified from pyrantel-treated individuals. The egg reappearance period (ERP) following treatment with ivermectin and pyrantel was investigated on two farms. The shortest ERP after ivermectin treatment was 8 weeks and after pyrantel was 5 weeks. We conclude that no substantial reversion to benzimidazole susceptibility had taken place, although these drugs have scarcely been used (<5%) in horses for the last 10 years. Pyrantel-resistant populations of cyathostomins are present on Swedish horse farms, but the overall efficacy of pyrantel is still acceptable.

The aim of this study was to investigate the suitability of a larval development assay (LDA) for the determination of anthelmintic resistance in cyathostomin nematode populations of the horse. In addition, comparison of results between geographic regions, types of horse establishment, and the use of anthelmintics in Sweden, was established. Seventy horse herds from different parts of Sweden were sampled, and strongyle eggs from the faeces of 54 of those were investigated by an LDA (DrenchRite). The following anthelmintics were tested: thiabendazole (TBZ), levamisole (LEV), ivermectin monosaccharide (IVM-MS), ivermectin aglycone (IVM-AG) and pyrantel (PYR). The LC50 values for TBZ and LEV were generally lower than those previously reported in other LDA studies on horse nematodes. This could be related to the infrequent use of these compounds for the past 20 years in Sweden. In this study, there was a great variation within and between assay plates that could not be explained. Still the LC50 values differed significantly between the regions for all anthelmintics, except for pyrantel. The highest LC50s were observed in parasite populations from the south of Sweden. There were no significant differences between riding schools and studs. Limitations of this technique exist, namely the lack of established cut-off values for susceptible and resistant populations and interpretation problems related to multi-species infections. Although there are advantages with LDA such as the possibility of testing several compounds simultaneously without interference with the deworming programmes on the farms, we conclude that LDA currently is not a reliable alternative to the faecal egg count reduction test (FECRT).

This study was conducted on a stud farm in Sweden to investigate the species composition of cyathostomins expelled in the faeces of horses after deworming using three different anthelmintic preparations. Twenty-seven horses excreting greater than or equal to200 strongyle eggs per gram faeces (EPG) were divided into three comparable groups and dewormed on day 0 with either of following compounds: 0.2 mg ivermectin per kg body weight (bw), 19 mg pyrantel pamoate per kg bw or 7.5 mg fenbendazole per kg bw. For each of the 3 days following anthelmintic treatment faeces was collected from individual horses and subsamples were fixed in formalin. Four days after the anthelmintic treatment all horses were re-treated with ivermectin and faeces was collected on day 5. Individual subsamples from each of the four sampling occasions were examined for cyathostomin nematodes. Sixty-three to 270 worms per horse were identified to the species level.The majority of the worms recovered were expelled during the first day from horses treated with ivermectin or pyrantel pamoate, and during the second day from horses treated with fenbendazole. Fifteen cyathostomin species were identified and the six most prevalent were Cylicocyclus nassatus, Cyathostomum catinatum, Cylicostephanus longibursatus, Cylicocyclus leptostomus, Cylicostephanus minutus and Cylicostephanus calicatus. These species composed 91% of the total burden of cyathostomins. The number of species found per horse ranged from 6 to 13, with an average of 9. No significant differences in species composition or distribution were found between the treatment groups. On day 5, i.e. 1 day after the last ivermectin treatment, 93% of the adult worms were recovered from horses in the fenbendazole group.This study showed that it was possible to identify cyathostomins expelled in faeces of dewormed horses, and that the most prevalent species corresponded to those found in autopsy surveys performed in other countries. (C) 2003 Elsevier B.V. All rights reserved.

Summary Faecal egg counts were performed during the first quarter of 1995 on samples from 1183 horses of varying breeds, ages and gender on 110 farms in 3 regions of Sweden. The majority of the horses had been treated with ivermectin or pyrantel when stabled the previous autumn. The risk of reinfection with strongyles between treatment and sampling was therefore considered to be minimal. Consequently, the results reflect primarily the occurrence of worms originating from inhibited strongyle larvae refractory to anthelmintic treatment. A total of 922 (78%) individuals were found to shed strongyle eggs, and 15 (14%) of the farms studied had at least one horse infected with Strongylus vulgaris. There was a significant, gradual decrease in faecal egg output in horses originating from the southern to the northern part of Sweden. Horses from studfarms showed significantly higher numbers of strongyle eggs/g faeces (EPG) as compared to those on other types of farms, such as riding‐schools and boarding stables. However, there was no correlation between herd size and mean herd EPG. The output of strongyle eggs was highest in horses aged 2 and 3 years. EPG values then declined with increasing age of the horse. Horses treated with ivermectin in the previous autumn showed significantly lower EPG values than untreated horses and those treated with pyrantel pamoate.

The egg output and humoral antibody response to scolex antigens of the equine tapeworm Anoplocephala perfoliata were monitored in naturally infected foals by an egg flotation/centrifugation method and an indirect enzyme-linked immunosorbent assay (ELISA). The study was performed on a stud farm in south-western Sweden between May 1994 and April 1995. Sequential blood and faecal samples were taken from 21 foals during their first summer on pasture and until tapeworm eggs were detected. Results were expressed separately for 10 and 11 foals born before and after the end of April 1994, respectively. Increased levels of antibodies were noticed from October and onwards in both groups whereas tapeworm eggs were detected in the faeces of all foals about 4 months later. The antibody response was similar in both groups but it was more pronounced in foals born before April 1994. All foals were treated in March 1995 with an oral paste formulation of pyrantel pamoate at a dose rate of 38 mg/kg bodyweight. Most animals responded to the anthelmintic treatment and one month later, tapeworm eggs were only detected in one out of the 18 foals examined one month after treatment. Thus, the treatment reduced the number of A. perfoliata egg positive horses by 94%. A concomitant decline in antibody levels was also observed. Western immunoblot analysis of sequential individual serum samples showed that at least 10 different scolex antigens in the molecular weight range 10–200 kDa were recognised. Banding intensities, especially of the 10 kDa, 35 kDa, 45 kDa and 66 kDa proteins were different in the sequential sera taken during the course of infection.