Multidrug resistance in Haemonchus contortus in sheep - can it be overcome?

Introduction: Gastrointestinal nematodes pose a threat to animal health and affect farmers by negatively impacting farm management. Material and Methods: The study was conducted on a sheep farm with suspected reductions in the efficacies of anthelmintics. Efficacy was determined using in vivo faecal egg count reduction, in vitro egg hatch (EHT) and larval development (LDT) tests. In the first phase, 60 sheep were equally split into six groups. Group 1 received the recommended dose of albendazole (ALB), group 2 received the same after fasting for 24 h, group 3 received the dose divided into two halves at 6 h intervals, group 4 received a double dose of ALB, and group 5 received the recommended dose of ivermectin (IVM). Group 6 served as a control. The second phase of the experiment had two groups: one treated with levamisole (LEV) and a control group. Faecal samples were collected from all sheep. Results: No reduction of egg output was observed in the groups treated with single, double, or divided doses of ALB, but one of 13.7-16.9% was noted in the fasting group. Efficacy in the IVM group ranged from 31.50 to 39.97%. The mean concentrations sufficient to prevent 50% of the eggs from hatching in the in vitro EHT and the mean concentrations in which the development of larvae to the L3 stage was inhibited by 50% in the LDT exceeded established thresholds for benzimidazoles and IVM. Haemonchus contortus was the only species identified after treatment. The LDT did not indicate the presence of resistance to LEV. All animals treated with LEV were negative for eggs 10 d after treatment. Conclusion: Resistance to ALB and IVM in Haemonchus contortus was confirmed. Alternative approaches to improve the efficacies of benzimidazole did not sufficiently increase the efficacy, but LEV was an efficient anthelmintic treatment.

Ashworthius sidemi Schulz, 1933 (Trichostrongylidae: Haemonchinae) in mountain ecosystems - a potential risk for the Tatra chamois Rupicapra rupicapra tatrica (Blahout, 1971/1972).

The Asian native Ashworthius sidemi is now in Europe, and several dozen years after its introduction, it is a widespread parasite of all wild cervids. For bovids, the nematode is a significant threat to the European bison (Bison bonasus) population and has also been found in mouflon (Ovis aries musimon). Our study aimed to assess the risk of infection for the endemic subspecies of northern (Alpine) chamois (Rupicapra rupicapra) - the Tatra chamois (R. r. tatrica), which has a critically endangered status. We conducted the investigation in the mountainous areas of Slovakia and Poland occupied by Tatra chamois (R. r. tatrica), Alpine chamois (R. r. rupicapra), red deer (Cervus elaphus) and roe deer (Capreolus capreolus). Animals (n = 93) shot during licensed hunting and killed in road accidents (roe deer, red deer), or which had died of natural causes (chamois) were post-mortem examined for the presence of Haemonchinae. The investigation confirmed the expansion of Ashworthius sidemi to high mountain regions via Cervidae. A. sidemi affected all of the examined roe deer and 90.0% of the red deer. As for the chamois, A. sidemi was found in one R. rupicapra originating from the Low Tatras, but not in any pure R. r. tatrica individuals living in the High and Western Tatras. Our work is the first confirmation of northern chamois infection with this alien and highly pathogenic blood-sucking nematode. Due to the important health hazard of A. sidemi infection for the Tatra chamois (R. r. tatrica), appropriate measures should be taken to reduce the possibility of parasite transmission between various cervid species living in the Tatra region, as well as the affected population of chamois and the pure Tatra chamois population inhabiting the higher parts of the mountains, constituting their natural habitat.

Morphometric and Molecular Analyses of Ostertagia leptospicularis Assadov, 1953 from Ruminants: Species Diversity or Host Influence?

Ostertagia leptospicularis Assadov, 1953 was formally described in roe deer Capreolus capreolus and has been reported in a wide range of ruminants, including other Cervidae, as well as Bovidae. Nematode specimens derived from various host species exhibit morphological similarity; however, some differences can be observed. It is unclear if this is due to the differential reaction of one nematode species in different host species (i.e., host-induced changes) or because of distinct nematode species in these hosts (i.e., species complex). This paper focuses on specimens resembling O. leptospicularis f. leptospicularis and its closely related species (Ostertagia ostertagi f. ostertagi) collected from various hosts. Morphometric and molecular techniques were applied to assess host-induced changes in nematode morphology and to clarify its systematic classification. There was an overall effect of host species on measurements of nematodes resembling O. leptospicularis (both males and females), but the distinctiveness of the specimens from cattle Bos taurus were highlighted. The results obtained may suggest that the specimens of O. leptospicularis from cattle in Germany and cervids in central Europe belong to different strains. Furthermore, nematodes from the cervid strain appear to circulate within particular host species, which can be seen in the stated morphological variations.

Coccidiosis ­ a problem in backyard rabbitries

This paper focuses on parasites of the genus Eimeria in rabbits kept in two backyard farms ­ from Poland (herd A) and Ukraine (herd B). The concentration McMaster technique and sporulation was applied to determine the level of infection, as well as to identify the species composition. The Eimeria sp. oocysts were present in all collected samples (prevalence = 100%), and four to nine coccidian species were identified in the herd A and B, respectively. Eimeria media was the most common species, and the presence of the most pathogenic E. intestinalis and E. flavescens was observed in both studied rabbitries. The results obtained suggest that coccidiosis may cause a serious risk for rabbits kept in backyard farms, therefore prevention guidelines in relation to the backyard breeding are crucial to be developed.

Gastrointestinal nematodes of European wild boar from distinct agricultural and forest habitats in Poland.

The study aimed to compare the gastrointestinal helminthofauna of free-ranging wild boars from arable lands and forests, which are the natural habitats for wild boar in Poland and further to investigate if wild boars living in agricultural environments could acquire helminths commonly detected in domestic pigs. In 2011-2014, a total of 57 wild boars were examined post-mortem for the presence of gastrointestinal nematodes. Altogether, all but two of the animals were infected, and seven nematode species were found. The mean infection burden was 68.9 parasites, ranging from 1 to 381 worms. In forest areas, Ascarops strongylina, Physocephalus sexalatus, and Globocephalus urosubulatus were common, whereas on arable lands, the animals were more frequently infected (P < 0.05) by Ascaris suum and Trichuris suis, which are parasites that commonly occur in domestic pigs. Oesophagostomum dentatum was observed only in wild boars on arable lands, and Bourgelatia diducta, which is alien to European suids, appeared irrespective of habitat type. These results show significant differences in parasite spectra among wild boars living in forests or arable lands in Poland and indicates the risks of parasite transfer from domestic pigs to free-ranging wild boars. Furthermore, in farmed game, organic farming, or in the case of agritourism farms, one should be aware of the risk of related animals acquiring new and alien parasite infections by being kept outdoors.

Heterakis isolonche Linstow, 1906 ­ a new nematode species found in ornamental pheasants in Poland

Studies were carried out on 25 pheasants belonging to 13 Phasianinae species of the Phasianidae family. The research material was collected from private breeders ­ both Polish and from abroad. The results confirm that the most frequent nematode is Heterakis gallinarum, while for the first time in the country ­ in two of the examined birds of Polish flocks ­ the occurrence of Heterakis isolonche was found.

Tarantobelus arachnicida (Nematoda: Rhabditida) invasion of exotic spiders in Poland

Nematodes of the Brevibuccidae family were stated among spiders of the Theraphosidae family (the South American species), which were bred in Poland. The first sign of infection was anorexia which led to gradually increasing lethargy progressed to a huddled posture. Additionally, a white discharge with nematodes between mouth and chelicerae was noted. All of the derived nematodes were morphologically identified and determined to the species Tarantobelus arachnicida. A molecular analysis covered amplification and sequencing of small subunit ribosomal RNA (18S rRNA). A post mortem examination demonstrated the presence of nematodes not only near the chelicerae, but also inside the intestine, hence the source of infection might be insects used as a food. The research showed that such kind of infection is an important disease, which poses a serious risk to the breeding spiders. To date there is no effective treatment, however, we demonstrated that usage of the Lugol's solution seems to be promising.

The occurrence and diversity of flies (Diptera) related to ruminant farming in southern Poland

The aim of the study was to determine the biodiversity of flies (Diptera) occurring in livestock buildings and in their direct proximity, in various systems of ruminants rearing. Flies were collected by net sweeping during the summer, in two cowsheds (one large-scale, and one traditional farm), and one specialist sheepfold, located in the Lesser Poland region of Poland. The insects were captured ­ both inside and outside the premises ­ three times (at 10:00, 12:00 and 14:00) on selected warm and sunny days of June, July and September, when animals were assumed to be the most harassed. A total of 18365 flies belonging to 13 families were obtained. The largest relative abundance of Diptera were from the Drosophilidae family (61.4%), and dominating Muscidae species ­ Musca domestica L., 1758 (19.2%) and Stomoxys calcitrans (L., 1758) (5.7%). More than half (55.5%) of all flies were captured in the morning, and their vast majority not inside a livestock building (28.2%), but in its close proximity (71.8%). This was particularly true of the large-scale cattle farm. The smallest number of insects, although with the highest biodiversity, were collected at the sheepfold. Regardless of their particular animal preferences, the insect species found were commonly occurring in different livestock sectors.

Host specificity and species diversity of the Ostertagiinae Lopez-Neyra, 1947 in ruminants: a European perspective.

BACKGROUND: Nematodes of the subfamily Ostertagiinae appear to be rather specific to a species or family of hosts, but some are observed in a wide variety of hosts. The nematode Ostertagia leptospicularis draws special attention due to its presence or absence among the same host species in different European countries. Therefore, this paper focuses mainly on the host specificity among nematodes of the subfamily Ostertagiinae. The second aim of this study is to assess the possibility of treating O. leptospicularis as an Ostertagia species complex. METHODS: Data were gathered from post-mortem examinations of domestic and wild ruminants (n = 157), as well as bibliographical references (n = 96), which were pooled and discussed. The research area was limited to European countries, hence the studied ostertagiine species are limited to native ones; likewise, the host species. Special emphasis was placed on the mean abundance values that allowed a typical host or hosts for each nematode species to be specified. Correspondence analysis was performed to confirm the stated host specificity. RESULTS: The analysis revealed that nematodes of this subfamily tend to use ruminants from a particular subfamily as their principal host. The results indicate that Ostertagia leptospicularis, similar to Teladorsagia circumcincta, may represent a potential species complex. This nematode, as the sole member of the subfamily Ostertagiinae, occurs in almost all representatives of the Bovidae subfamily, as well as in the Cervidae. CONCLUSIONS: Despite the stated narrow host specificity, the results obtained may suggest that O. leptospicularis is not strongly connected to any host or is comparably associated with a very wide and diverse group of hosts (Cervidae, Bovidae). The Ostertagia complex may have particular cryptic species or strains typical for any individual host or group of hosts. Such a conclusion requires further investigations on a wider scale.

Cystic echinococcosis in Poland: genetic variability and the first record of Echinococcus granulosus sensu stricto (G1 genotype) in the country.

Cystic echinococcosis is one of the most important zoonotic diseases affecting humans and livestock worldwide, and is endemic in Poland. A set of six isolates on larval stages of Echinococcus granulosus sensu lato tapeworms collected from three humans, two pigs and one sheep from Polish foci of CE was examined by DNA sequencing of two mitochondrial genes (cox1, rrnS). The results demonstrated the presence of E. canadensis and E. granulosus sensu stricto in the investigated hydatid cysts. The former species was found in all five isolates from pigs and humans derived from central Poland. In a sheep hydatid cyst originating from Lesser Poland Voivodeship in southern Poland, E. granulosus s. s. (G1 genotype) was identified. This is the first report of an unambiguously autochthonous infection with E. granulosus s. s. in Poland. The global distribution and host affiliations of the commonly occurring G1 microvariant with nucleotide change 56C/T in cox1, detected here in Polish sheep, are discussed. The finding that sheep harboured E. granulosus s. s. may have important consequences for developing effective hydatid control programmes in Poland due to its longer maturation rate in dogs compared with E. canadensis G7. This may lead to greater expenditures for purchasing anthelmintics to provide an appropriate dosing regime in sheep-raising areas of the country.

Cephenemyia stimulator (Diptera) infection in roe deer (Capreolus capreolus) from Kraków area, southern Poland.

UNLABELLED: The aim of the study was to determine the prevalence of botfly (Diptera: Oestridae) larvae infection in roe deer populations (Capreolus capreolus) in the Kraków area on the basis of necropsy and questionnaire surveys. Hunters were surveyed about the age and sex of hunted animals, and the origin of their habitat. All parasite specimens were identified to species in the laboratory. The parasites were found in the nasal cavities, esophagus, and larynx of male roe deer aged 3-8 years, living in forest habitats. The level of infection was relatively low (13%), with the intensity ranging from 1 to 10 larvae per host. Although no fly larvae were found during the dissection of roe deer carcasses, the parasites received from the hunters were identified as Cephenemyia stimulator botflies. These findings are supported by the evidence drawn from the questionnaires completed by the hunters. KEY WORDS: parasites, Cephenemyia stimulator, Capreolus capreolus, necropsy, questionnaire study, Poland.

Benzimidazole resistance in the ovine Haemonchus contortus from southern Poland - coproscopical and molecular findings.

UNLABELLED: Anthelmintic resistance within nematodes has become a very common issue, however, the data about its occurrence in the gastrointestinal nematodes of sheep in Poland are very limited. This study was carried out in order to evaluate the presence of benzimidazole resistance in highly pathogenic Haemonchus contortus by means of parasitological and molecular techniques. The research represents the first Polish attempt to confirm the presence of a mutation at codon 200 of the ß-tubulin isotype 1 gene in H. contortus by PCR-RFLP. The occurrence of this mutation indicates the risk of ineffective benzimidazole treatment, nowadays commonly used for parasite control in Poland. The resistant parasites were detected by means of FECRT in a sheep flock (30 individuals) from southern Poland. To confirm the resistance on the molecular level, primers designed according to the sequences available in Genebank were used to detect the mutation. Unfortunately, as the PCR product was shorter than required (403 bp), further analyses are needed. The obtained results may indicate the high variability within the parasite population. Hence, it is essential to adapt the reaction conditions to our geographic strain of the nematode - and further analyses are required. KEY WORDS: Haemonchus contortus, benzimidazole resistance, PCR, Poland.

Endoparasites of exotic ungulates from the Giraffidae and Camelidae families kept ex situ.

Giraffes and camels are popular attractions at zoological gardens. In order to present the diversity of parasites infecting exotic ungulates from zoos, faecal samples from three giraffes and six camels from both the Silesian Zoological Garden in Chorzów, and Kraków Zoological Garden, were examined. The research was carried out over a ten-month period in 2013 and 2014. In total, 100 faecal samples from 18 animals were analysed with the use of the McMaster method. Moreover, coccidian oocysts were incubated to investigate their development and larvoscopic examination was conducted to detect the presence of nematode species. Giraffes were infected with coccidia from the genus Eimeria, and gastrointestinal nematodes from the Strongylida order, and Trichuris and Aonhotheca genera. One male giraffe was uninfected. The level of infection in giraffes was low when compared to camels kept in both of the zoos. Limited contact with other animal species contributed greatly to the lower level of infection in camels from Kraków Zoo than those from Chorzów, which were kept in the same enclosure as alpacas and Shetland ponies.

Lungworm (Nematoda: Protostrongylidae) infection in wild and domestic ruminants from Malopolska region of Poland.

The conducted study has focused on domestic, as well as wild ruminant species. The post mortem examination was carried out on 68 animals, including three wild species: roe deer (Capreolus capreolus) (25 indyviduals), red deer (Cervus elaphus) (6), fallow deer (Dama dama) (5) and two domestic: sheep (Ovis aries) (14) and cattle (Bos taurus) (18). Some of the species have also been investigated in the field by the coproscopical analyses. The faecal samples from roe deer (27), fallow deer (20), red deer (36) and moose (Alces alces) (10) were collected from the environment, while from sheep (10) and goat (Capra hircus) (10)--per rectum. Based on the obtained results the following values were calculated: prevalence, mean intensity and intensity range. The post mortem examination did not reveal pulmonary nematodes neither in domestic nor in wild ungulates, however, the larvae of aforementioned parasites were often stated in the stool samples taken from the environment. All wild species, except fallow deer were infected. Consequently, six species of lungworms have been identified. The first stage larvae of Varestrongylus capreoli occurred in 11 samples of roe deer and Varestrongylus alces in one moose. The larvae of Elaphostrongylus cervi were found in 19 red deer and Varestrongylus sagittatus in 3. Furthermore, Elaphostrongylus alces larvae were noted in 6 moose. Within domestic ruminants only one sheep and two goats were infected by Muellerius capillaris.

Parasite infections in red deer Cervus elaphus from Krakow area, southern Poland.

The study describes the parasitofauna of red deer Cervus elaphus from Kraków area (southern Poland). The study was done in years 2011-2012 and included altogether 6 animals. Five deer originated from the forest areas (hunted) and one came from the urban site (it died in a road accident). In addition to post-mortem examination, coproscopic analyses with sporulation were performed to define Eimeria species diversity. The deer were infected with four species of nematodes: Ashworthius sidemi, Spiculopteragia spiculoptera, Oesophagostomum venulosum and O. sikae, and two species of coccidia: E. elaphi and E. austriaca. The level of parasitie infections was low, but the results indicate the important role of deer as possible transmitter of A. sidemi to other ungulates in examined area.

Zatorska goose - a subject of parasitological research.

The aim of the study was to determine the level of gastrointestinal parasites in a native breed of geese - Zatorska goose - based on coproscopic testing. Faecal samples were collected from 90 young geese in three age groups (5, 7 and 9 weeks old) in 2014. The geese were kept indoors on deep litter and pastured from spring to autumn. The area of the pastures around the buildings where the geese grazed was about 1 hectare, divided into quarters for different age groups. Before grazing, the birds were dewormed with fenbendazole (Fenbenat powder 4%, Naturan). As additional treatment for coccidiosis, coccidiostats were added to the feed. The study was conducted using the McMaster quantitative method with centrifugation (flotation liquid: NaCl and glucose). The birds were shown to be infected with coccidia and nematodes. The prevalence of Eimeria sp. infection (mean 40%) and the number of oocysts per gram of faeces (reaching 5,300 OPG) were highest in the youngest age group of geese. The level of Amidostomum anseris infection was similar in the three age groups, with prevalence from 40% to 50% (nematode egg output ranged from 50 to 350 eggs per gram of faeces, EPG). Capillaria anatis was observed only in 5- and 7-week-old geese.

The Alice - "Follow the White Rabbit" - parasites of farm rabbits based on coproscopy.

The aim of the study, conducted in the years 2011-2013, was to determine the level of gastrointestinal parasites infection in New Zealand White rabbits, kept at the Experimental Station of the University of Agriculture in Krakow. The study showed rabbits protozoan infection with the genus Eimeria, belonging - based on the sporulation method - to the following species: E. magna, E. media, E. perforans, E. stiedae and E. irresidua. The highest prevalence of infection, as well as the intensity of oocysts output (OPG - oocysts per gram of faeces), was noted for E. magna and E. media - respectively 31.4 % (19477.3 OPG), and 40.0 % (14256.07 OPG). The infection of rabbits with Eimeria spp. differed significantly between years. With regard to oocysts output, the level of infection was strongly connected with the age of rabbits, being higher in young animals. However, the range of infection was highest among adults. Among nematodes, Passalurus ambiguus pinworm was regularly found (prevalence reached 21.9%), other species - Trichuris leporis, and Graphidium strigosum were rarely noted. The overall infection with nematodes did not differ between years. Similarly, as in the case of Eimeria older individuals were more often infected by nematodes. We observed some trends in parasite oocysts/eggs output; the protozoan oocysts were recorded more often in faecal samples collected in the evenings, whereas the nematodes eggs occurred frequently in the mornings. This situation may be related to the phenomenon of coprophagy occurring in the mammals of Lagomorpha order. The results of the study indicate that especially coccidiosis constitute permanently throughout the years an important problem in the rabbitry examined.

An identification key for the five most common species of Metastrongylus.

Species of the Metastrongylus genus, the lung nematodes of pigs that require an intermediate host (earthworm) to complete their cycle, pose a potential risk to both livestock and humans. This parasite which can result in lung pathology and mixed infections with other pathogens (e.g. viruses) can be fatal to pigs. Although this genus is distributed worldwide, there are no classification keys for identifying this common parasite species. In this work, we take advantage of parasitological surveys of wild boar (Sus scrofa) in northern and central Spain and southern Poland to develop a morphological identification key for the five most common Metastrongylus species (Metastrongylus apri, Metastrongylus pudendotectus, Metastrongylus salmi, Metastrongylus confusus and Metastrongylus asymetricus). In addition, we provide the first record of M. confusus in Spain, probably unidentified until now due to the lack of appropriate identification keys. We hope that this user-friendly identification key will enable parasitologists and veterinary practitioners to avoid further misclassifications of Metastrongylus species.

Presence of blood-sucking mesostigmatic mites in rodents and birds kept in pet stores in the Cracow area, Poland.

The aim of the study was to investigate the occurrence of zoonotic arthropod parasites in small animals sold in selected pet stores in the Cracow area. The research was conducted in seven pet stores, keeping a total of six species of rodents and three species of birds. In two shops, two species of mites of the order Mesostigmata were detected on the animals and in their surrounding: Dermanyssus gallinae, the poultry red mite, and Ornithonyssus bacoti, the rat mite. Both observed species of mites may be harmful to animals, as well as to people working in the shops or potential pet owners. This study discusses the possible origin of the parasites and their importance to human health.

Herbal formulations as feed additives in the course of rabbit subclinical coccidiosis.

Two simultaneous experiments were carried out in a breeding farm of New Zealand White rabbits (Oryctolagus cuniculus f. domesticus) to determine the feasibility of replacing coccidiostats with garlic and oregano preparation. The research took place during June and July, the period of the greatest threat of coccidiosis caused by Eimeria spp. (Apicomplexa: Eimeriidae). In one investigation, 40 rabbits aged 1-3 months were divided into four groups of ten animals: Group A being a control which received no coccidiostats in feed, Group B receiving the coccidiostat Baycox in water once at weaning, Group C receiving the coccidiostat robenidine in feed, and group D receiving herbal extracts in feed. In the second trial, six mated females were allocated equally to three groups analogous to A, C, and D above during pregnancy and lactation. Bulk stool samples were collected from each group of rabbits at weekly intervals for coproscopic analysis, and the production results of the animals were recorded. In the young rabbits, both the faecal coccidia oocyst counts and body weight gains were more favourable in group D than the remaining groups. Also, the female rabbits of group D were the least infected. The results demonstrate that garlic and oregano feed additives exert a positive influence on the level and course of coccidia infection, with regard to maintaining a good level of animal productivity, and these herbal extracts appear to have potential value in coccidiosis prophylaxy.