Corrigendum: A revision of the trichostrongylid nematode Cooperia Ransom, 1907, from deer game: recent integrative research confirms the existence of the ancient host-specific species Cooperia ventricosa (Rudolphi, 1809).

[This corrects the article DOI: 10.3389/fvets.2024.1346417.].

A revision of the trichostrongylid nematode Cooperia Ransom, 1907, from deer game: recent integrative research confirms the existence of the ancient host-specific species Cooperia ventricosa (Rudolphi, 1809).

The trichostrongylid roundworms of the genus Cooperia, which are important in veterinary medicine, currently comprise 19 valid species that parasitize the small intestine of both free-living and domestic ruminants. Only four Cooperia spp. have been reported in Europe, namely C. oncophora, C. punctata, C. curticei and C. pectinata. In 2018-2022, 25 red deer (Cervus elaphus) and 30 sika deer (Cervus nippon) of both sexes and various ages from several remote locations in the Czech Republic were parasitologically examined. Intestinal nematodes of the genus Cooperia were found only in two northern regions. Using the globally recognized key book on trichostrongylid nematodes, they were preliminarily identified as C. pectinata. However, a molecular analysis of cox2 and ITS rDNA gene sequences revealed that Cooperia sp. parasitizing Czech deer is a separate taxon that is more closely related to C. oncophora than to C. pectinata. A subsequent morphological analysis and literature survey confirmed the independence of deer Cooperia sp., which is similar but not identical to bovid C. pectinata. Previous long-term correct identifications of bovid C. pectinata and misidentifications of deer Cooperia species were caused by a fundamental error in the key book mentioned above. Interestingly, the ancient trichostrongylid nematode Strongylus ventricosus from the type host red deer (Cervus elaphus) shot near Greifswald (Germany) was described by Rudolphi in 1809. Rudolphi's type material (one male and four females) was deposited in the Museum für Naturkunde (Berlin). Later, the ancient species S. ventricosus was taken as a synonym for various Cooperia spp. Our current re-examination of the type male indicated that there is a relatively good agreement with our new material from Czech deer regarding the most important characteristics of S. ventricosus (i.e., the shape and size of the male spicules); however, Rudolphi's type material is in rather poor condition. The suggested resurrection of the deer Cooperia sp. in this study as Cooperia ventricosa (Rudolphi, 1809) requires verification by collecting and analyzing new nematode material from the type locality near Greifswald.

Significance of Intestinal Helminth Infection and Animal Sex for Mercury Concentrations in Two Rodent Species.

We compared the effects of animal gender, species, and intestinal helminth burden on mercury concentrations in rodents. Total mercury concentrations were determined in the liver and kidney tissues of 80 small rodents (44 yellow-necked mice, Apodemus flavicollis, and 36 bank voles, Myodes glareolus) captured in the Ore Mountains (northwest Bohemia, Czech Republic). Overall, 25/80 (32%) of animals were infected by intestinal helminths. The differences in mercury concentration between rodents infected and not infected with intestinal helminths were not statistically significant. Statistically significant differences in mercury concentrations were found only between voles and mice (that were not infected with intestinal helminths). This suggests the differences may be associated with host genetics. Apodemus flavicollis body tissues had significantly lower (P=0.01) mean Hg concentrations (0.032 mg/kg) than Myodes glareolus (0.279 mg/kg), provided that animals were not infected by intestinal helminths; if the animals were infected by intestinal helminths, the difference between both groups was insignificant. The effect of gender in this study was significant only for voles (without helminth infection); for mice (either with or without helminth infection) the differences between genders were not significant. Myodes glareolus males had significantly lower (P=0.03) Hg concentrations in liver and kidney tissues (0.050 mg/kg) than Myodes glareolus females (0.122 mg/kg). These results reveal the importance of considering species and gender when evaluating mercury concentrations.

Effects of excessive dietary zinc or zinc/cadmium and tapeworm infection on the biochemical parameters in rats.

The main objective of this study was to determine which biochemical blood parameters can serve as indicators of Zn or Zn/Cd burden and tapeworm infection. This study was performed on 44 Wistar male rats during a 6-week period, when rats were or were not fed a zinc/cadmium rich diet and were or were not infected with tapeworms (Hymenolepis diminuta). Total protein, albumin, urea, glucose, triacylglycerols, non-esterified fatty acids, cholesterol, alkaline phosphatase, aspartate aminotransferase, uric acid, Mg, Ca, P and Zn levels were analysed. Control rats with tapeworm infection had significantly higher (p ≤ 0.05) total protein, urea and phosphorus concentrations than did rats unaffected by any experimental factor. Rats given overdoses of zinc lactate exhibited significantly lower glucose levels than did the other rats, especially those infected with tapeworms. Low glucose level in uninfected rats indicate a Zn overdose; high doses of zinc lactate likely decrease levels of glucose via cortisol, which is released during stress. Rats fed the Zn/Cd hyperaccumulating plant Arabidopsis halleri and infected with tapeworms had significantly higher (p ≤ 0.01) cholesterol and urea levels but lower zinc, triacylglycerol, and alkaline phosphatase levels than did rats fed the same diet but free of tapeworms. The increase of alkaline phosphatase level in uninfected rats may indicate both Zn/Cd burden and rat liver damage. Overall, this study not only supports the theory that H. diminuta can serve as a promising model for helminth therapy of the host mammal but also confirmed that this tapeworm is capable to protect somehow the host organism from the harmful effects of heavy metals.

Setaria cervi (Filarioidea, Onchocercidae) undressing in ungulates: altered morphology of developmental stages, their molecular detection and complete sequence cox1 gene.

This work introduces new morphological and molecular information on the filaroid nematode Setaria cervi (Rudolphi, 1819) obtained from 13 infected game ungulates out of 96 dissected. The hosts comprised the following: a single moose (Alces alces), ten red deer (Cervus elaphus) and two sika deer (Cervus nippon) originating from the western and northern regions of the Czech Republic. Based on the complete sequences of the gene encoding mitochondrial cytochrome c oxidase subunit 1 (cox1), all 20 females and four males belonged to the species S. cervi. We detected three developmental female stages (adult fertile females, juvenile L5 females and L4 female larvae) differing in size and some morphological traits as the subtle structure of peribuccal crown and shape and features of tail knob. Such differences were described in detail for the first time. The phylogenetic relationships within the family Onchocercidae have been evaluated using new information on the cox1 sequence of S. cervi (maximum likelihood method, GTR + I + G model). In accordance with the latest phylogenetic studies, the present analysis confirmed the ancient separation of the subclass Setariinae from the remaining two onchocercid lineages Dirofilariinae and Onchocerinae.

Effects of tapeworm infection on absorption and excretion of zinc and cadmium by experimental rats.

The main objective of this study was to determine how rat tapeworms affect the excretion of zinc and cadmium through rat feces. Male rats (Rattus norvegicus var. alba) were divided into four groups, and the experiment was conducted over a 6-week period. The control groups (00; 0T) were provided with a standard ST-1 rodent mixture and received 10.5 mg of Zn/week. Groups P0 and PT were fed a mixture supplemented with the hyperaccumulating plant Arabidopsis halleri at a dosage of 123 mg Zn/week and 2.46 mg Cd/week. Groups 0T and PT were infected with the rat tapeworm (Hymenolepis diminuta). Fecal samples were collected 24 h post exposure. Zinc and cadmium concentrations in rat feces were analyzed using inductively coupled plasma optical emission spectrometry. Tapeworm presence decreased the amount of metals excreted through the feces of the host throughout the entire experiment, with the exception of 1 week (control group). No statistically significant differences between zinc excretion rates in the control groups (00 and 0T) were detected at any time throughout the experiment. A statistically significant difference between zinc excretion rates (p < 0.05) in the exposed groups (P0 and PT) was detected in 2 of the 6 monitored weeks. Group PT excreted significantly less cadmium (p < 0.01) than group P0 did in three of the 6 weeks. Overall, our results indicate that tapeworms are able to influence the excretion of metals by their host. Tapeworms accumulate metals from intestinal contents. It is not clear whether tapeworms carry out this process before the host tissues absorb the metals from the intestines or the tapeworms accumulate metals excreted from the body of the host back to the intestines. Most likely, it is a combination of both phenomena.

Long-term occurrence of Trichuris species in wild ruminants in the Czech Republic.

The aim of this study was to identify Trichuris species in wild ruminants from 32 localities in the Czech Republic using morphological and molecular methods (ITS1-5.8S RNA-ITS2 region polymorphisms). Trichurids were obtained from 176 wild ruminants (roe deer, sika deer, red deer, fallow deer and mouflons) that were culled between 2009 and 2017. Trichuris discolor is the predominant trichurid of all of the above-mentioned wild ruminants, whereas Trichuris ovis was identified less frequently in roe deer, fallow deer, sika deer and mouflons. Red deer were parasitised exclusively by T. discolor. Young hosts under 1 year of age were more intensively infected by trichurids than were adults (χ2 = 32.02, p = 0.00). Trichurid prevalence results obtained through coprological methods and those based on parasitological dissections differed significantly (χ2 = 16.26, p = 0.00). The regression analysis indicated that the eggs per gram (EPG) threshold (20 EPG) was exceeded only if the host was parasitised by more than 7 trichurid females. Full concordance between the positive results obtained by the coprological methods and those obtained via direct dissections was achieved when the number of trichurid females per host exceeded 51.

How tapeworm infection and consumption of a Cd and Zn hyperaccumulating plant may affect Cu, Fe, and Mn concentrations in an animal-a plant consumer and tapeworm host.

This study evaluated the effects of a hyperaccumulator plant (Arabidopsis halleri), containing surplus of cadmium (Cd) and zinc (Zn) and being an admixture to the rat feed, on concentrations of copper (Cu), iron (Fe), and manganese (Mn) in the tissues of experimental rats infected/uninfected with the tapeworm (Hymenolepis diminuta). Male Wistar rats were divided into three groups (00, P0, and PT); the P0 and PT animals were fed a standard mixture for rats (ST-1) supplemented with the plant A. halleri at a weekly Zn and Cd dosage of 123 and 1 mg, respectively. Moreover, rats from the group PT were infected with the tapeworm. The group 00 served as control animals fed only ST-1 having no tapeworm infection. Rats were euthanized after 6 weeks, and Cu, Fe, and Mn levels were determined in rat and tapeworm tissues. The results indicated that both the consumption of hyperaccumulator plant and/or presence of tapeworms did have significant effect on Cu, Fe, and Mn concentrations in the host tissues. Concentrations of all the elements were higher in the rat liver and partially kidneys than in the tapeworms, and the concentrations of Cu, Fe, and Mn were affected by the consumption of Cd/Zn hyperaccumulator plants. Particularly, Fe concentrations in all rat tissues were significantly increased by consumption of A. halleri while decreased by the presence of tapeworms. Overall, the consumption of a Cd/Zn hyperaccumulator plant and tapeworm infection cause an imbalance in Cu, Fe, and Mn concentrations in the tissues of a consumer (experimental rats).

Reliable molecular differentiation of Trichuris ovis and Trichuris discolor from sheep (Ovis orientalis aries) and roe deer (Capreolus capreolus) and morphological characterisation of their females: morphology does not work sufficiently.

The main aim of the study was to evaluate associations between morphological variability of Trichuris females from sheep and roe deer and their rDNA polymorphism in whipworm populations from the Czech Republic. The results introduced the use of new molecular markers based on the internal transcribed spacer (ITS)1-5.8S RNA-ITS2 region polymorphisms, as useful tools for the unambiguous differentiation of congeners Trichuris ovis and Trichuris discolor. These markers revealed both parasites in roe deer and in sheep; however, T. ovis females predominated in sheep while T. discolor females occurred mostly in roe deer. Additional analysis of ITS1-5.8 rRNA-ITS2 discovered the genetic uniformity of the analysed T. discolor but high haplotype variation of T. ovis. Simultaneously, molecularly designated female individuals of both species were categorised into four morphotypes (MT) on the basis of morphology of genital pore area. MT1 and MT4 (vulvar opening on everted vaginal appendage/on visible cuticular bulge) occurred only in T. ovis, MT2 (uneverted vagina-vulvar opening without any elevation) was identified only in T. discolor and MT3 (transient type of vulvar opening on a small swelling) was observed in both species. Statistical analysis of biometric data confirmed that morphology of vulva is not a reliable marker for the species determination. On the basis of the ITS1-5.8S RNA-ITS2 region variability, we carried out a phylogenetic analysis (maximum likelihood method, Hasegawa-Kishino-Yano model) which showed that T. ovis haplotypes from the Czech Republic and Ireland and T. discolor haplotypes from the Czech Republic, Spain, Iran and Japan are sister OTUs.

Can the Hyperaccumulating Plant Arabidopsis halleri in Feed Influence a Given Consumer Organism (Rattus norvegicus var. alba)?

Zinc and cadmium concentrations in rat (Rattus norvegicus var. alba) tissues were analyzed by inductively coupled plasma optical emission spectrometry. Rats were fed the zinc and cadmium hyperaccumulating plant, Arabidopsis halleri. When compared to the control group, a Cd increase in all tissues (liver, kidneys, small intestine, spleen, testes, muscle), with the exception of bone tissue was observed. In comparison to the control group, the kidneys, liver and small intestine contained 375, 162, and 80 times more Cd, respectively. Differences between zinc concentrations in rats fed with A. halleri and those of the control group were significant only in the small intestine and kidney tissues. Results suggest using the hyperaccumulating plant A. halleri as a feed stresses the consumer organism not through its Zn content, but through its Cd content.

Heavy metal concentrations in the small intestine of red fox (Vulpes vulpes) with and without Echinococcus multilocularis infection.

Heavy metal (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) levels in red fox small intestine samples with or without Echinococcus multilocularis infection were studied. The red foxes were taken from the open countryside of northwest Bohemia (CR). Red foxes with E. multilocularis infection had lower levels of toxic metals (Cd, Pb); cadmium levels in infected foxes (0.0052 mg/kg) were twice as low as in uninfected foxes (0.0106 mg/kg). This was the same case for lead: 0.0288 mg/kg infected red foxes (inf.) and 0.0413 mg/kg uninfected (uninf.). Conversely, red foxes with E. multilocularis infection yielded higher concentrations in comparison to their uninfected counterparts: Cr (0.0087 mg/kg uninf. and 0.0116 mg/kg inf.), Cu (0.2677 mg/kg uninf. and 0.3205 mg/kg inf.), Fe (6.46 mg/kg uninf. and 10.89 mg/kg inf.), Mn (0.1966 mg/kg uninf. and 0.2029 mg/kg inf.), Ni (0.0415 mg/kg uninf. and 0.064 mg/kg inf.) and Zn (16.71 mg/kg uninf. and 20.25 mg/kg inf). This could support the hypothesis that tapeworms are able to absorb toxic heavy metals from the host body into their tissues, as well as to modify other element concentrations in the host body.

Arrested development of experimental Cyathostominae infections in ponies in Czech republic.

Nine ponies, aged 12 months at the time of infection, were randomly allocated to one of three groups. Each pony was infected with 30,000 infective cyathostome larvae stored 4 weeks under the natural environmental conditions of the Czech Republic. Horses of Group A were infected with larvae conditioned from August 16 to September 12 (the first phase of the experiment). Horses of Group B were inoculated with larvae conditioned from October 10 to November 4 (the second phase of the experiment). In the third phase, horses from Group C received larvae stored under natural conditions (from 8.11 to 18.11.2011). The larvae were then kept under laboratory conditions (5 °C, the light reduction according to the nature condition). In the first and second phase of the experiment, 98.57% and 95.48% of adult nematodes were found, respectively. However, only 29.07% of adults were found in the third phase of experiment, and the proportion of arrested larvae (EL3) was 67.73%. The share of these larvae in the cecum during last phase of the experiment comprised 92.59% of all cyathostomes.

Is the tapeworm able to affect tissue Pb-concentrations in white rat?

The effect of gastrointestinal helminths on Pb accumulation in the host body is ambiguous. A laboratory experiment with Rattus norvegicus/Hymenolepis diminuta model was conducted to determine Pb toxicokinetics in a terrestrial host-parasite system. The ET-AAS or ICP-OES techniques were used to determine Pb concentrations (CPb) in both tapeworms and host tissues (kidney, liver, bone, testes, muscle and intestinal wall). Concerning the entire host-parasite system, the highest CPb were detected in H. diminuta. Rat kidneys and bone were the only two tissues whose mean Pb levels were lower in parasitized animals than they were in non-infected subjects after both levels of exposure. At low Pb exposure, parasitization slightly changed the Pb toxicokinetics in the host body. However, with respect to tissue at the same exposure level, no significant differences were detected between the parasitized and non-parasitized animals and no significant correlations were found between CPb in tapeworms and those of host tissues. The results of this study indicate that H. diminuta does not protect rat from elevated Pb exposure even if tapeworm accumulates a higher portion of ingested Pb dose compared with that of the most Pb-loaded host soft tissue. The portion of Pb dose accumulated in H. diminuta correlates positively with parasite biomass.

A 4-years monitoring of Hypoderma diana in horses from the Czech Republic.

The occurrence of Hypoderma diana (Diptera: Hypodermatidae) in a herd of 15 horses was observed during a 4-year period (2010-2013) in northwest Bohemia. During this period, infestation by the warble fly H. diana increased from 12.5% in 2010 to 53.3% in 2013. Nodules were observed on the neck, back, stomach, legs, and head. The incidence of H. diana was probably influenced by the presence of a specific host mouflon (Ovis orientalis musimon).

Importance of fish gender as a factor in environmental monitoring of mercury.

Total mercury concentrations were determined in the gonadal tissues of 15 female and 10 male European perch (Perca fluviatilis) from one location of the stream "Jevanský potok" located about 30 km from Prague (Czech Republic). Tissue samples were frozen at -26 °C in polypropylene containers until further processing, which was carried out using an Advance Mercury Analyser (single purpose atomic absorption spectrometer). Mercury concentrations were present in all analysed gonad samples, and ranged from 2.3 to 12.7 µg/kg wet weight. However, we determined a mean Hg concentration (9.45 µg/kg) in male gonads that was 2.4 times greater than that of female gonads (3.9 µg/kg). This is an important finding when taking into account fish sex in environmental pollution monitoring (especially for mercury contamination).

Faecal excretion dynamic during subacute oral exposure to different Pb species in Rattus norvegicus.

Faecal excretion is a basic means of detoxification upon ingestion of Pb-contaminated feed. In order to determine a time course of Pb elimination after oral exposure to two different forms of this heavy metal (lead acetate vs. phyto-bound Pb), a feeding study was carried out in experimental rats using the Pb phyto-hyperaccumulator Pistia stratiotes as a model diet. The effect of starvation on Pb excretion was further studied in rats that were fed plant material. Twelve Pb doses (7 µg Pb/1 g BW) were administered orally over a 5-week period. Faeces samples were collected 24 and 72 h post-exposure. Inductively coupled plasma optical emission spectrometry and electrothermal absorption spectrometry methods were used for determination of heavy metal concentrations. Up to 53 % of ingested Pb was rapidly eliminated from the exposed rats via faeces within 24 h after exposure. Faecal excretion in exposed rats differed significantly when compared to that of the control group. Fasting before exposure reduced Pb excretion by up to 50 %. Faecal excretions of both examined Pb forms exhibited almost identical patterns. Considerable differences were revealed concerning total excretion levels; lead acetate was excreted in amount greater extent than those of phytobound Pb. Results of our study suggest that Pb forms occurring in the P. stratiotes tissues are absorbed through the gastrointestinal tract to a greater extent than Pb from lead acetate. Therefore, higher portions of ingested Pb can be available for potential accumulation in tissues of exposed subjects.

Cephenemyia stimulator and Hypoderma diana infection of roe deer in the Czech Republic over an 8-year period.

A survey of naso-pharyngeal and subcutaneous myiasis affecting roe deer (Capreolus capreolus) was conducted in the Czech Republic over an 8-year period (1999-2006). A total of 503 bucks and 264 does from six hunting localities were examined. The sampling area comprised predominantly agricultural lowlands and a mountain range primarily covered by forest. Since 1997, the deer have been treated each winter across the board with ivermectin (150 mg/kg, CERMIX® pulvis, Biopharm, CZ). Parasites found were the larvae of Hypoderma diana and Cephenemyia stimulator. There were no significant differences in warble fly infection among captured animals in the individual hunting localities. Overall, 146 (28.8%) of 503 animals (bucks) were infected with Cephenemyia stimulator larvae; body size of the second instar larva reached 13-18 mm. The prevalence ranged from 16.1 to 42.9% per year, and the mean intensity from 6 to 11 larvae per animal. Additionally, a total of 264 roe deer (does) were examined for H. diana larvae, and 77 (29.1%) were found to be positive; body size of the second instar larva reached 17 mm. The prevalence ranged from 18.8 to 50.0% per year, and the mean intensity from 13 to 22 larvae per animal. The results showed that the bot flies, Cephenemyia stimulator as well as H. diana, are common parasites in roe deer in the Czech Republic, and that through the help of treatment (ivermectin), it is possible to keep parasite levels low. The body weights of infected and non-infected H. diana deer did not differ significantly.

Trichomonas spp. in pigeons: detection by OSOM Trichomonas Rapid Test.

The efficacy of the OSOM Trichomonas Rapid Test (developed for rapid diagnosis of human Trichomonas vaginalis) in detection of Trichomonas spp. in pigeons (Columba livia) was investigated. Two oral cavity swabs were taken from 50 farm pigeons. Cultivation in Diamond Trichomonas medium was used as a reference method. According to a morphological determination, Trichomonas gallinae was the only protozoan found; however, no further molecular analysis was conducted. The OSOM Trichomonas test was positive in 39 oral swabs. In comparison with the cultivation method three samples were identified as false negative and one as false positive. Test specificity and sensitivity were established as 93% and 90%, respectively. Using Cohen's Kappa, the concordance between the two testing methods was found to be strong (kappa = 0.7506, 95% CI = 0.5162-0.9850). The OSOM Trichomonas test is not able to distinguish between Trichomonas species; however, results suggest that the test is suitable for the rapid detection of Trichomonas spp. infection in pigeons.

Effect of Acanthocephalus lucii infection on total mercury concentrations in muscle and gonads of fish host (Perca fluviatilis).

The samples from 13 perches (Perca fluviatilis) - muscle with skin and bones; fish gonads; and acanthocephalan parasites were analysed for mercury (Hg). Hg concentrations were present in all analysed samples. There were found no statistically significant difference in Hg concentration in fish tissues between perches either with or without infection by the acanthocephalan parasite, Acanthocephalus lucii). In this study there was no evidence that acanthocephalan worms accumulate mercury from hosts. For this reason, A. lucii is not a suitable bioindicator for mercury pollution.

Concentrations of Zn, Mn, Cu and Cd in different tissues of perch (Perca fluviatilis) and in perch intestinal parasite (Acanthocephalus lucii) from the stream near Prague (Czech Republic).

We monitored concentrations of Cd, Cu, Mn and Zn in acantocephalan parasites (Acanthocephalus lucii) and its final host (Perca fluviatilis). The concentrations in parasites were found to be significantly higher than those found in the muscle, gonads and liver of fish host. The bioaccumulation factor values (BF=C(parasite)/C(host muscle)) were 194, 24.4, 2.2 and 4.7 for Cd, Cu, Mn and Zn, respectively. This suggests a benefit for the host due to the high accumulation of toxic cadmium.