Exotic ticks removed from German travelers.

We report the finding of five nymphs and three adult ticks attached to German tourists while traveling the American continents. All eight specimens were morphologically identified and confirmed genetically using the 16S rRNA gene and screened for Rickettsia spp. infections. Five tick species were identified: one Amblyomma mixtum nymph from Ecuador, one Amblyomma varium nymph from Colombia, three Amblyomma coelebs nymphs from Costa Rica, one Amblyomma americanum male from the USA, one Dermacentor andersoni female and one D. andersoni male from Canada. Tick-borne microorganisms screening using the pan-Rickettsia-PCR resulted in two positive and six negative ticks. The A. mixtum nymph was positive for Rickettsia amblyommatis, while the D. andersoni female was positive for Rickettsia peacockii.

Morphological changes in plasma-exposed poultry red mites (Dermanyssus gallinae) using high-resolution video camera and optical coherence tomography (OCT).

Dermanyssus gallinae, the poultry red mite (PRM), is a hematophagous temporary ectoparasite that causes serious economic losses and animal health impairment on laying hen farms worldwide. Control is limited by the parasite's hidden lifestyle, restrictions on the use of chemical acaricides and the development of resistance against certain drug classes. As a result, research was conducted to explore alternative control methods. In recent years, atmospheric pressure plasma has been increasingly reported as an alternative to chemical acaricides for pest control. This physical method has also shown promising against PRM under laboratory conditions. However, the detailed mechanisms of action have not yet been elucidated. In the present study, the effects of cold atmospheric pressure plasma on PRM were investigated using digital videography and optical coherence tomography (OCT), an imaging technique that visualizes the topography of surfaces and internal structures. Digital videography showed that a redistribution of the contents of the intestinal tract and excretory organs (Malpighian tubules) occurred immediately after plasma exposure. The body fluids reached the distal leg segments of PRM and parts of the haemocoel showed whiter and denser clumps, indicating a coagulation of the haemocoel components. OCT showed a loss of the boundaries of the hollow organs in transverse and sagittal sectional images as well as in the three-dimensional image reconstruction. In addition, a dorso-ventral shrinkage of the idiosoma was observed in plasma-exposed mites, which had shrunk to 44.0% of its original height six minutes after plasma exposure.

Anthelmintic Activities of Extract and Ellagitannins from Phyllanthus urinaria against Caenorhabditis elegans and Zoonotic or Animal Parasitic Nematodes.

The aerial parts of Phyllanthus urinaria are used in traditional medicine in West Africa against helminthiasis, but their anthelmintic potential has not been evaluated until now. Within the current study, a hydroacetonic extract (AWE) and fractions and isolated ellagitannins from P. urinaria were, therefore, tested in vitro against Caenorhabditis elegans and the larvae of the animal parasites Toxocara canis, Ascaris suum, Ancylostoma caninum, and Trichuris suis. Compounds 1:  - 13: , mainly representing ellagitannins, were isolated using different chromatographic methods, and their structures were elucidated by HR-MS and 1H/13C-NMR. AWE exerted concentration-dependent lethal effects (LC50 of 2.6 mg/mL) against C. elegans and inhibited larval migration of all animal parasites tested (T. suis L1 IC50 24.3 µg/mL, A. suum L3 IC50 35.7 µg/mL, A. caninum L3 IC50 112.8 µg/mL, T. canis L3 IC50 1513.2 µg/mL). The anthelmintic activity of AWE was mainly related to the polar, tannin-containing fractions. Geraniin 1: , the major ellagitannin in the extract, showed the strongest anthelmintic activity in general (IC50 between 0.6 and 804 µM, depending on parasite species) and was the only compound active against A. caninum (IC50 of 35.0 µM). Furosin 9: was least active despite structural similarities to 1: . Among the parasites tested, Trichuris suis L1 larvae turned out to be most sensitive with IC50 of 0.6, 6.4, 4.0, 4.8, and 2.6 µM for geraniin 1: , repandusinic acid A 3: , punicafolin 8: , furosin 9: , and phyllanthusiin A 10: , respectively.

Filarial infections in lemurs: Evidence for a wide geographical distribution and low host specificity among lemur species.

The relevance of emerging infectious diseases continues to grow worldwide as human activities increasingly extend into formerly remote natural areas. This is particularly noticeable on the island of Madagascar. As closest relatives to humans on the island, lemurs are of particular relevance as a potential origin of zoonotic pathogen spillover. Knowledge of pathogens circulating in lemur populations is, however, very poor. Particularly little is known about lemur hemoparasites. To infer host range, ecological and geographic spread of the recently described hemoparasitic nematode Lemurfilaria lemuris in northwestern Madagascar, a total of 942 individuals of two mouse lemur species (Microcebus murinus [n = 207] and Microcebus ravelobensis [n = 433]) and two rodent species (the endemic Eliurus myoxinus [n = 118] and the invasive Rattus rattus [n = 184]) were captured in two fragmented forest landscapes (Ankarafantsika National Park and Mariarano Classified Forest) in northwestern Madagascar for blood sample examination. No protozoan hemoparasites were detected by microscopic blood smear screening. Microfilaria were present in 1.0% (2/207) of M. murinus and 2.1% (9/433) of M. ravelobensis blood samples but not in rodent samples. Internal transcribed spacer 1 (ITS-1) sequences were identical to an unnamed Onchocercidae species previously described to infect a larger lemur species, Propithecus verreauxi, about 650 km further south. In contrast to expectations, L. lemuris was not detected. The finding of a pathogen in a distantly related host species, at a considerable geographic distance from the location of its original detection, instead of a microfilaria species previously described for one of the studied host species in the same region, illustrates our low level of knowledge of lemur hemoparasites, their host ranges, distribution, modes of transmission, and their zoonotic potential. Our findings shall stimulate new research that will be of relevance for both conservation medicine and human epidemiology.

Successful long-term control of poultry red mite (Dermanyssus gallinae) infestations in floor-kept laying hens via integrated pest management-a case report.

This case report describes the successful control of poultry red mite [PRM] (Dermanyssus gallinae) infestations in an experimental laying hen house via a combined use of cleaning and disinfection measure, the preventive application of a synthetic silica-based acaricide and frequent mite monitoring. The high number of PRM in the laying hen house was reduced by 99.8% by treatment with fluralaner (Exzolt®, MSD Animal Health Unterschleißheim, Germany; 0.5 mg/kg body weight via drinking water twice, 7 days apart). After the laying hens were removed, the hen house was dry-cleaned, wet-cleaned and disinfected. After drying, synthetic amorphous silica (Fossil Shield® instant white, Bein GmbH, Eiterfeld, Germany) was applied as a preventive measure before the hen house was restocked with pullets for two housing periods of 58 and 52 weeks. Over these periods (i.e. more than 2 years), no PRM was detected during mite monitoring at two-week intervals via tube traps and visual monitoring. This result therefore suggests that the combined use of appropriate chemical and physical prevention measures within an integrated pest management regime can be successfully used for the long-term control of PRM. This could reduce the use of acaricidal drugs, thereby helping maintain their efficacy.

Evaluation of a commercial coproantigen immunoassay for the detection of Toxocara cati and Ancylostoma tubaeforme in cats and Uncinaria stenocephala in dogs.

Coproantigen immunoassays (IDEXX Fecal Dx® antigen tests) were evaluated for their ability to identify Toxocara cati and Ancylostoma tubaeforme infections in cats and Uncinaria stenocephala infection in dogs. Five cats were experimentally infected with 500 embryonated eggs of T. cati, eight cats with 500 third-stage larvae (L3) of A. tubaeforme and seven dogs with 500 L3 of U. stenocephala. In addition to the three coproantigen tests, the course of infection was monitored by a combined sedimentation-flotation method with ZnSO4 as flotation medium (specific gravity: 1.28-1.30) and a modified McMaster method in case of copromicroscopically positive samples. Eggs of T. cati were first observed between 28 and 54 days post infection (dpi) in four of the five infected cats. In these four cats, positive roundworm coproantigen signals were obtained between 16 and 44 dpi. Positive coproantigen signal always preceded egg observations, but the interval varied between 6 and 30 days. Hookworm-specific positive coproantigen signals were detected in seven of the eight A. tubaeforme infected cats between 10 and 52 dpi, while consecutive egg excretion was observed in three cats between day 26 and 54 pi. Of these three, coproantigen signal preceded egg observation by 12 to 24 days. Four cats had positive coproantigen results in the absence of egg excretion, and one cat never achieved a positive result for egg or coproantigen. In six of seven U. stenocephala infected dogs, infection was confirmed by copromicroscopy between 16 and 24 dpi as well as for hookworm coproantigen between 10 and 14 dpi. Coproantigen signal was detected prior to egg observation by 2 to 14 days. No cross-reactions between the roundworm, hookworm und whipworm tests occurred in study animals. The results of this study demonstrate the ability of the coproantigen tests to detect the common roundworm and hookworm infections in cats and U. stenocephala infections in dogs as well as the ability to detect the prepatent stage of infection.

Short-time cold atmospheric pressure plasma exposure can kill all life stages of the poultry red mite, Dermanyssus gallinae, under laboratory conditions.

In the present study, the acaricidal effects of cold atmospheric pressure plasma treatment on poultry red mites of different developmental stages have been investigated under laboratory conditions using a dielectric barrier discharge system. A total of 1890 poultry red mites and 90 mite eggs, respectively, were exposed to the plasma under various parameter settings with a single plasma pulse generated using the gas mixture of the ambient air at atmospheric pressure. The results showed that all developmental stages of the poultry red mite could be killed by cold atmospheric pressure plasma treatment. Plasma exposure to mite eggs resulted in a complete 100% hatch inhibition regardless of the parameter settings. Post-exposure mortality rates of larvae, nymphs and adults showed significant differences after utilization of plasma at 10 W for 1.0 s. In addition, the mortality rate increased with progressing time after plasma exposure. An average mortality rate of 99.7% was observed after 12 h in all mites exposed to plasma, regardless of the selected plasma parameter, developmental stage, and nutritional status of the mites. Cold atmospheric pressure plasma has an acaricidal effect on all developmental stages of Dermanyssus gallinae, suggesting that it could be developed to an effective method for the control of poultry red mites in laying hen husbandry.

Molecular characterization of Cysticercus tenuicollis isolates from sheep in the Nile Delta, Egypt and a review on Taenia hydatigena infections worldwide.

The predator­prey-transmitted cestode Taenia hydatigena infects a wide range of definitive and intermediate hosts all over the world. Domestic and sylvatic cycles of transmission are considered as well. The parasite has considerable economic importance, particularly in sheep. Here, the molecular characters of T. hydatigena cysticerci in sheep from the Nile Delta, Egypt were investigated for the first time. For this purpose, 200 sheep carcasses and their offal were inspected at the municipal abattoir, Dakahlia governorate, Egypt. Cysticerci of T. hydatigena were collected and molecularly characterized employing the mitochondrial 12S rRNA gene. Cysticerci were found in 42 (21%) sheep, mostly attached to the omenti, mesenteries and livers. After molecular confirmation, nine isolates were sequenced displaying six different haplotypes. Analysis of the T. hydatigena 12S rRNA nucleotide sequences deposited in GenBank revealed 55 haplotypes out of 69 isolates, displaying high haplotype (0.797) and low nucleotide (0.00739) diversities. For the Tajima D neutrality index, a negative value (−2.702) was determined, indicating the population expansion of the parasite. Additionally, global data summarized in this study should be useful to set up effective control strategies against this ubiquitous parasite.

A new ELISA and western blot technique based on recombinant TES antigen and/or larval antigen for the detection of toxocariasis in humans.

Serological antibody detection by enzyme-linked immunosorbent assay (ELISA)- and immunoblot-based methods constitutes the best indicator of human Toxocara infection. Nevertheless, the availability of serological tests, particularly western blots (WB), evaluated for sensitivity and specificity is limited. Therefore, an Anti-Toxocara-ELISA immunoglobulin g (IgG) prototype (Proto-ELISA) and an Anti-Toxocara-Westernblot (IgG) prototype (Proto-WB) were evaluated by testing 541 human sera pre-determined for Toxocara infection by an established in-house Anti-Toxocara-ELISA (IH-ELISA). To evaluate sensitivity and specificity of the newly developed ELISA and WB prototypes, results were compared to IH-ELISA and a commercial WB (Com-WB). Compared to the IH-ELISA, a sensitivity of 93.1% (229/246) and a specificity of 94.6% (279/295) of the Proto-ELISA with a Cohen's κ of 0.88 were obtained. The sensitivity of the Proto-WB was 76.7% (240/313) and specificity was 99.6% (227/228) with a Cohen's κ of 0.73 compared to those of Com-WB. A comparison to the IH-ELISA revealed 91.5% (225/246) sensitivity and 94.6% (279/295) specificity of the Proto-WB with a Cohen's κ of 0.86. Cross-reactivity was observed for some samples positive for Ascaris and Trichinella spp. in the Proto-ELISA, Proto-WB and Com-WB. Overall, the evaluated ELISA and WB prototypes showed high sensitivity and specificity, indicating high reliability of these newly developed tests.

Responsiveness of fecal immunoglobulin A to HPA-axis activation limits its use for mucosal immunity assessment.

The assessment of mucosal immunity as a component of animal health is an important aspect for the understanding of variation in host immunity, and its tradeoff against other life-history traits. We investigated immunoglobulin A (IgA), the major type of antibody associated with mucosal immunity, in relation to changes in parasitic burden following anthelminthic treatment in noninvasively collected fecal samples in a semi-free ranging group of Barbary macaques (Macaca sylvanus). We measured IgA in 340 fecal samples of fourteen females and nine males. As IgA has been found to be responsive to stressors, we also related fecal IgA (fIgA) levels to fecal glucocorticoid metabolites (fGCM) measured in the same samples as part of a previous study. We found a high variability within and between individual fIgA levels over time. Running generalized additive mixed models, we found that fIgA levels were higher in males than in females, but did not change in response to the anthelmintic treatment and the resulting reduction in worm burden. Instead, fIgA level changes were significantly correlated to changes in fGCM levels. Our findings indicate that due to the strong responsiveness of fIgA to HPA-axis activity, the measurement of fIgA may have certain limitations with respect to reflecting gastrointestinal parasitic burden. Moreover, the responsiveness of fIgA to stressors interferes with the interpretation of IgA levels in fecal samples as a measure of mucosal immunity, at least in our study population of the Barbary macaques.

Tick-borne zoonoses and commonly used diagnostic methods in human and veterinary medicine.

Around the world, human health and animal health are closely linked in terms of the One Health concept by ticks acting as vectors for zoonotic pathogens. Animals do not only maintain tick cycles but can either be clinically affected by the same tick-borne pathogens as humans and/or play a role as reservoirs or sentinel pathogen hosts. However, the relevance of different tick-borne diseases (TBDs) may vary in human vs. veterinary medicine, which is consequently reflected by the availability of human vs. veterinary diagnostic tests. Yet, as TBDs gain importance in both fields and rare zoonotic pathogens, such as Babesia spp., are increasingly identified as causes of human disease, a One Health approach regarding development of new diagnostic tools may lead to synergistic benefits. This review gives an overview on zoonotic protozoan, bacterial and viral tick-borne pathogens worldwide, discusses commonly used diagnostic techniques for TBDs, and compares commercial availability of diagnostic tests for humans vs. domestic animals, using Germany as an example, with the aim of highlighting existing gaps and opportunities for collaboration in a One Health framework.

Predators in northern Germany are reservoirs for parasites of One Health concern.

Urbanisation and invasion of wildlife into urban areas as well as human leisure activities create diverse wildlife-domestic animal-human interfaces, increasing the risk of (zoonotic) parasite spillover from sylvatic to domestic and synanthropic cycles. This study investigated the endo- and ectoparasite fauna, emphasising on parasites of One Health Concern, of the most common predators in northern Germany between November 2013 and January 2016. Eighty red foxes (Vulpes vulpes), 18 stone martens (Martes foina) and nine raccoon dogs (Nyctereutes procyonoides) were available for the study. Overall, 79 (73.8%) of the examined predators (n=107) harboured at least one endoparasite. The most frequently detected endoparasites in red foxes were Toxocara canis (43.8% positive individuals), Capillaria spp. (36.3%), Alaria alata (25.0%), Echinococcus multilocularis (26.3%) and Uncinaria stenocephala (25.0%). Furthermore, Toxascaris leonina, Trichuris vulpis, Taenia ssp., Mesocestoides spp. and coccidian oocysts were observed. The endoparasite species richness in raccoon dogs was comparable to red foxes, while in stone martens, only Capillaria spp. were found. Muscle digestion for detection of Trichinella spp. and antigen testing for Giardia spp. did not show positive results. Ectoparasite analyses revealed infestations with ticks species of the genus Ixodes as well as Dermacentor reticulatus. Scabies mites were not present in digested skin samples, while Demodex spp. mites were observed by faecal flotation in one red fox. Furthermore, fleas (Archaeopsylla erinacei and Chaetopsylla globiceps) were observed in the fur of red foxes, while lice were not present in any predator species. However, infestation frequency with ectoparasites was with 19.2% generally low in available predator skins (n=99). Overall, the present study showed that predators in northern Germany serve as reservoirs for parasites of One Health concern, with four of the five most frequent endoparasites being zoonotic, highlighting the need of parasite surveillance in wildlife predators in order to implement measures avoiding spillovers to domestic animals and humans.

Bovine Babesiosis Diagnosed in Formalin-Fixed, Paraffin-Embedded Tissues by Using In Situ Hybridization.

Bovine babesiosis, caused by Babesia divergens, is in general a rare disease in Europe. Nonetheless, local outbreaks can cause severe economic damage, and postmortem identification represents a diagnostic challenge. During a recent outbreak in May 2018 in northern Germany, 21 animals of a herd of 150 cattle died within 40 days having had clinical signs of fever and hemoglobinuria. Gross examination of 4 of the 21 deceased animals revealed a tick infestation, jaundice, and dark brown staining of urine and kidneys. Histologically, there were iron-positive deposits, hyperplasia of the red pulp of the spleen, and centrilobular necrosis of hepatocytes. In several locations, small basophilic granules suggestive of intraerythrocytic parasites were visible in hematoxylin-eosin- and Giemsa-stained sections. Peripheral blood smears from a living cow from the herd and polymerase chain reaction (PCR) of feeding ticks revealed B. divergens infection. In situ hybridization (ISH) was applied on formalin-fixed, paraffin-embedded (FFPE) tissue of the necropsied cattle to confirm babesiosis in these animals postmortem. Digoxigenin-labeled DNA probes were generated based on a specific nucleotide sequence for B. divergens, obtained by PCR and sequencing of DNA isolates from infected Ixodes ricinus ticks from deceased cattle. ISH using these probes allowed postmortem diagnosis of B. divergens infection in routinely fixed FFPE tissues.

Feline lungworms in Greece: copromicroscopic, molecular and serological study.

Aelurostrongylus abstrusus (Nematoda, Metastrongyloidea) causes verminous pneumonia in cats worldwide. This study evaluated the seroprevalence of A. abstrusus antibodies in 220 stray and free-roaming cats from insular (Mykonos, Crete, Skopelos) and continental (Thessaloniki, Attica) Greece. The results were compared with morphological and biomolecular identification of first-stage larvae (L1) in faeces. Positive cats were observed in all 5 areas: 13/97 (13.4%), 1/32 (3.1%), 7/26 (26.9%), 3/18 (16.7%) and 5/47 (10.6%) cats tested positive for A. abstrusus L1 by Baermann examination, and 33/97 (34.0%), 7/32 (21.9%), 6/26 (23.1%), 3/18 (16.7%) and 11/47 (23.4%) were seropositive, in Mykonos, Crete, Skopelos, Thessaloniki and Attica, respectively. Troglostrongylus brevior L1 were found in 12/97 (12.4%), 3/26 (11.5%) and 2/47 (4.3%) cats from Mykonos, Skopelos and Attica respectively. Six of the 220 cats (2.7%), i.e. 4/97 (4.1%) from Mykonos and 2/26 (7.7%) from Skopelos, shed L1 of both A. abstrusus and T. brevior. Sixty samples were ELISA-positive (27.3%, 95% CI: 21.5-33.7%), of which 21 (35%) tested copromicroscopically positive (19 monospecific infections and 2 mixed with Troglostrongylus brevior), and 5 were positive for T. brevior L1 only. Among seronegative cats (n = 140), L1 of A. abstrusus were additionally detected in 8 (5.7% out of 140) cats (i.e. 4 monospecific infections and 4 mixed with T. brevior), and in 6 (4.3% out of 140) cats, L1 of T. brevior as monospecific infection were detected. This study confirms the presence of lungworms in Greece and suggests that the number of cats infected with/exposed to metastrongylids is higher than detected by faecal examinations.

Genome-wide associations and functional gene analyses for endoparasite resistance in an endangered population of native German Black Pied cattle.

BACKGROUND: Gastrointestinal nematodes (GIN), liver flukes (Fasciola hepatica) and bovine lungworms (Dictyocaulus viviparus) are the most important parasitic agents in pastured dairy cattle. Endoparasite infections are associated with reduced milk production and detrimental impacts on female fertility, contributing to economic losses in affected farms. In quantitative-genetic studies, the heritabilities for GIN and F. hepatica were moderate, encouraging studies on genomic scales. Genome-wide association studies (GWAS) based on dense single nucleotide polymorphism (SNP) marker panels allow exploration of the underlying genomic architecture of complex disease traits. The current GWAS combined the identification of potential candidate genes with pathway analyses to obtain deeper insights into bovine immune response and the mechanisms of resistance against endoparasite infections. RESULTS: A 2-step approach was applied to infer genome-wide associations in an endangered dual-purpose cattle subpopulation [Deutsches Schwarzbuntes Niederungsrind (DSN)] with a limited number of phenotypic records. First, endoparasite traits from a population of 1166 Black and White dairy cows [including Holstein Friesian (HF) and DSN] naturally infected with GIN, F. hepatica and D. viviparus were precorrected for fixed effects using linear mixed models. Afterwards, the precorrected phenotypes were the dependent traits (rFEC-GIN, rFEC-FH, and rFLC-DV) in GWAS based on 423,654 SNPs from 148 DSN cows. We identified 44 SNPs above the genome-wide significance threshold (pBonf = 4.47 × 10- 7), and 145 associations surpassed the chromosome-wide significance threshold (range: 7.47 × 10- 6 on BTA 1 to 2.18 × 10- 5 on BTA 28). The associated SNPs identified were annotated to 23 candidate genes. The DAVID analysis inferred four pathways as being related to immune response mechanisms or involved in host-parasite interactions. SNP effect correlations considering specific chromosome segments indicate that breeding for resistance to GIN or F. hepatica as measured by fecal egg counts is genetically associated with a higher risk for udder infections. CONCLUSIONS: We detected a large number of loci with small to moderate effects for endoparasite resistance. The potential candidate genes regulating resistance identified were pathogen-specific. Genetic antagonistic associations between disease resistance and productivity were specific for specific chromosome segments. The 2-step approach was a valid methodological approach to infer genetic mechanisms in an endangered breed with a limited number of phenotypic records.

Neuroinvasion of Toxocara canis- and T. cati-larvae mediates dynamic changes in brain cytokine and chemokine profile.

BACKGROUND: Neurotoxocarosis (NT) is induced by larvae of the dog or cat roundworm (Toxocara canis or T. cati) migrating and persisting in the central nervous system of paratenic hosts, including humans, and may be accompanied by severe neurological symptoms. Host- or parasite-induced immunoregulatory processes contribute to the pathogenesis, but detailed data on pathogenic mechanisms and involvement of signalling molecules during cerebral Toxocara species infections are scarce. METHODS: To elucidate alterations in immunomodulatory mediator pattern, comprehensive multiplex bead array assays profiling comprising 23 different cytokines and chemokines were performed during the course of T. canis- and T. cati-induced NT. To this end, cerebra and cerebella of experimentally infected C57Bl/6 J mice serving as paratenic host models were analysed at six different time points (days 7, 14, 28, 42, 70 and 98) post infectionem (pi). RESULTS: Brain-body mass ratios of T. canis and T. cati-infected mice were significantly lower than those of the uninfected control group at day 14 pi, and also at day 28 pi for T. canis-infected mice. Both infection groups showed a continuous decrease of pro-inflammatory cytokine concentrations, including TNF-α, IFN-γ, GM-CSF and IL-6, in the cerebrum over the course of infection. Additionally, T. canis but not T. cati-induced neurotoxocarosis was characterised by significantly elevated levels of anti-inflammatory IL-4 and IL-5 in the cerebrum in the acute and subacute phase of the disease. The higher neuroaffinity of T. canis led to a prominent increase of eotaxin and MIP-1α in both the cerebrum and cerebellum, while in T. cati-infected mice, these chemokines were significantly elevated only in the cerebellum. CONCLUSIONS: The direct comparison of T. canis- and T. cati-induced NT provides valuable insights into key regulatory mechanisms of Toxocara species in paratenic hosts. The cerebral cyto-/chemokine milieu is shifted to a predominantly anti-inflammatory immune response during NT, possibly enabling both survival of the parasite and the neuroinfected paratenic host. Alteration of eotaxin and MIP-1α concentrations are congruent with the higher neuroaffinity of T. canis and species-specific tropism of T. canis to the cerebrum and T. cati to the cerebellum.

Allele substitution and dominance effects of CD166/ALCAM gene polymorphisms for endoparasite resistance and test-day traits in a small cattle population using logistic regression analyses.

The study investigated the effects of four single-nucleotide polymorphisms (SNPs) in the activated leukocyte cell adhesion molecule (ALCAM) gene on liver fluke (Fasciola hepatica) infections (FH-INF), gastrointestinal nematode infections (GIN-INF) and disease indicator traits [e.g. somatic cell score (SCS), fat-to-protein ratio (FPR)] in German dual-purpose cattle (DSN). A genome-wide association study inferred the chip SNP ALCAMc.73+32791A>G as a candidate for F. hepatica resistance in DSN. Because of the crucial function of ALCAM in immune responses, SNPs in the gene might influence further resistance and performance traits. Causal mutations were identified in exon 9 (ALCAMc.1017T>C) and intron 9 (ALCAMc.1104+10T>A, ALCAMc.1104+85T>C) in a selective subset of 94 DSN cows. We applied logistic regression analyses for the association between SNP genotypes with residuals for endoparasite traits (rINF-FH, rGIN-INF) and estimated breeding values (EBVs) for test-day traits. The probability of the heterozygous genotype was estimated in dependency of the target trait. Allele substitution effects for rFH-INF were significant for all four loci. The T allele of the SNPs ALCAMc.1017T>C and ALCAMc.1104+85T>C was the favourable allele when improving resistance against FH-INF. Significant allele substitution for rGIN-INF was only found for the chip SNP ALCAMc.73+32791A>G. We identified significant associations between the SNPs with EBVs for milk fat%, protein% and FPR. Dominance effects for the EBVs of test-day traits ranged from 0.00 to 0.47 SD and were in the direction of improved resistance for rFH-INF. We estimated favourable dominance effects from same genotypes for rFH-INF and FPR, but dominance effects were antagonistic between rFH-INF and SCS.

Histopathological characterization of Toxocara canis- and T. cati-induced neurotoxocarosis in the mouse model.

Infective larvae of Toxocara canis and T. cati, the common roundworms of dogs and cats, may invade the central nervous system of paratenic hosts, including humans, causing neurotoxocarosis (NT). Previous studies on NT in the model organism "mouse" have indicated distinct differences between T. canis and T. cati regarding larval migration patterns as well as the severity of clinical symptoms and behavioural alterations. The objective of the present study was to provide an extensive characterization of the underlying histopathological alterations, comparing T. canis- and T. cati-induced changes in different brain areas over the course of murine infection. Four histological sections of five brains each of T. canis- and T. cati-infected as well as uninfected C57Bl/6 mice were investigated 7, 14, 28, 42, 70 and 98 days post infection (dpi), while brains of T. cati-infected and control mice were also available 120 and 150 dpi. In addition to haematoxylin-eosin and luxol fast blue-cresyl violet staining, immunohistochemistry was employed to study microglia/macrophage cell morphology and to detect accumulation of ß-amyloid precursor protein (ß-APP) as an indicator of axonal damage. Haemorrhages, eosinophilic vasculitis and activated microglia/macrophages were detected in both infection groups starting 7 dpi, followed by eosinophilic meningitis in cerebra as from 14 dpi. Overall, little differences in the proportion of animals affected by these alterations were found between the two infection groups. In contrast, the proportion of animals displaying ß-APP accumulation was significantly higher in the T. canis than T. cati group as from 28 dpi regarding the cerebrum as well as at 98 dpi regarding the cerebellum. In T. canis-infected mice, myelinophagic microglia/macrophages ("gitter cells") appeared as from 14 dpi, whereas these were first observed at 70 dpi in T. cati-infected animals. The proportion of animals displaying demyelination and/or gitter cells in the cerebrum was significantly higher in the T. canis than T. cati group as from 28 dpi, and at 28 and 42 dpi regarding the cerebellum. Earlier and more severe neurodegeneration during T. canis- than T. cati-induced NT, especially in the cerebrum, may explain the differences in behavioural alterations observed in previous studies. In addition to differences in larval migration preferences, immunological processes may contribute to these patterns, which warrant further investigation.

Ixodes inopinatus in northern Germany: occurrence and potential vector role for Borrelia spp., Rickettsia spp., and Anaplasma phagocytophilum in comparison with Ixodes ricinus.

In 2014, a new tick species, Ixodes inopinatus, was described, which is closely related to Ixodes ricinus. So far, I. inopinatus has been found in Tunisia, Morocco, Spain, Portugal, Romania, Austria, and southern Germany. No data is yet available regarding occurrence of I. inopinatus in northern Germany and the potential role of I. inopinatus as a vector for tick-borne pathogens. Therefore, 3845 DNA samples from Ixodes ticks collected for prevalence studies on Borrelia spp., Rickettsia spp., and Anaplasma phagocytophilum during the years 2010-2015 in the northern German cities of Hamburg and Hanover were differentiated into I. ricinus or I. inopinatus by sequencing a part of the 16S rRNA gene. In total, 4% (137/3845) of the sequenced ticks were assigned to the species I. inopinatus and 96% (3708/3845) to I. ricinus. The prevalence of Borrelia spp., Rickettsia spp., and A. phagocytophilum DNA in I. inopinatus was 34% (46/137), 46% (63/137), and 3% (4/137), respectively, whereas the prevalence of these bacteria in I. ricinus was 25% (919/3708), 47% (1729/3708), and 4% (135/3708), respectively. Compared with I. ricinus, significantly more I. inopinatus ticks tested positive for Borrelia. To the best of our knowledge, this is the first report of I. inopinatus in northern Germany. Detection of the DNA of Borrelia spp., Rickettsia spp., and A. phagocytophilum in questing I. inopinatus indicates a potential role of this tick species as a vector of these pathogens, which needs to be confirmed by transmission experiments.

Helminth infections of wild European gray wolves (Canis lupus Linnaeus, 1758) in Lower Saxony, Germany, and comparison to captive wolves.

This study aimed to investigate the endoparasite fauna of wild European gray wolves, which are currently recolonizing Germany. In total, 69 fecal samples of wild wolves were collected in Lower Saxony, Germany, from 2013 to 2015, analyzed by the sedimentation-flotation and McMaster techniques and compared to previous results on captive European Gray wolves living in zoological gardens in Germany. In addition to coproscopy, taeniid-positive samples from wild as well as captive wolves were differentiated by amplification and sequencing of small subunit ribosomal RNA (SSU rRNA) and NADH dehydrogenase 1 (nad1) gene fragments. Missing Taenia krabbei SSU rRNA reference sequences were generated from two T. krabbei specimens. Overall, 60.87% (42/69) of wild wolve samples were microscopically positive for at least one of seven egg types. Capillaria/Eucoleus spp. showed the highest frequency (31.88% [22/69]), followed by Taeniidae (21.74% [15/69]), Ancylostomatidae (20.29% [14/69]), Alaria alata (15.94% [11/69]), Toxocara canis (13.04% [9/69]), and Toxascaris leonina and Trichuris vulpis (each 5.80% [4/69]). Amplification of SSU rRNA was successful for 7/15 Taeniidae-positive samples from wild and 20/39 samples from captive wolves, revealing T. hydatigena in two and 14 samples, respectively. Taenia krabbei was detected in two further samples of wild and three samples of captive wolves, while for the remaining samples, no differentiation between T. serialis/T. krabbei was possible. Echinococcus spp. were not detected. Sequence comparisons revealed that the SSU rRNA gene fragment was not suitable to differentiate between T. serialis and T. krabbei. Therefore, the use of this fragment alone cannot be recommended for species identification in future studies.