The vicuna (Vicugna vicugna) as a natural host of Dictyocaulus filaria in Peru.

Lungworm infection, or verminous pneumonia, is a parasitic disease that causes serious problems in small and large ruminants. Despite the fact that nematodes of the genus Dictyocaulus in cattle and sheep are the main cause of this disease, there are few studies on the natural infections of South American camelids. For this reason, this study aims to report the natural infection by Dictyocaulus filaria in vicunas (Vicugna vicugna) for the first time. During a shearing season (chaku) in Cuzco, Peru, two accidentally killed adult vicunas were submitted to the IVITA-Marangani research center in Cuzco for their respective necropsies. The tracheas of both vicunas had numerous nematodes, as seen during the necropsy. The nematodes were collected in 70% ethanol and were morphologically identified as D. filaria. Likewise, the DNA of six nematodes was extracted, and the ITS2 region and the 28S rRNA gene were amplified and sequenced. The nucleotide sequences of both genetic markers were up to 100% identical with previously reported D. filaria DNA sequences found in the goat yearlings from Turkey, sheep from Iran, Turkey, and India, and the argali from Uzbekistan, which confirmed the morphological diagnosis. This finding represents the first molecular confirmation of a natural D. filaria infection in a South American camelid. It will be necessary to carry out future studies to know the current situation of verminous pneumonia in domestic and wild South American camelids and to know the negative effects of the disease on them.

Larvoscopic study on Dictyocaulus sp. in the faeces of beef cattle in northeastern Brazil / Estudo larvoscópico de Dictyocaulus sp. em fezes de bovinos de corte do nordeste do Brasil

Abstract The lungworm Dictyocaulus viviparus has an important role in cattle health and productivity worldwide, since infections can lead to substantial economic losses. Despite its importance, few studies investigating the epidemiological aspects of infection by this parasite have been conducted. The aim of this study was to report the occurrence of lungworm infection in beef cattle herds reared in an area of livestock production in the northeastern region of Brazil. From September 2020 to August 2021, monthly fecal samples (n = 493) were collected from 46 beef cattle. Among all the animals assessed, lungworm larvae were detected in 8.7% (4/46). None of them presented any clinical sign suggestive of infection by lungworm parasites. Twenty larvae were retrieved, with the minimum number (n = 1) detected in October and December, and the maximum number (n = 13) in November. These presented a mean length of 363 μm (± 28.65 μm) and mean width of 19 μm (± 1.03 μm), and were morphologically similar to Dictyocaulus sp.. This study reports the occurrence of this parasite in this livestock production area. Lastly, local veterinarians need to be aware of inclusion of this parasite in the differential diagnosis of other respiratory infections in beef cattle.

Resumo O parasito pulmonar Dictyocaulus viviparus tem um papel importante na saúde e produtividade bovina em todo o mundo, uma vez que infecções podem levar a perdas econômicas. Além de sua importância, poucos estudos que investigam aspectos epidemiológicos das infecções por esse parasito têm sido realizados. Objetivou-se, nesta pesquisa, relatar a ocorrência de infecção por nematódeos pulmonares em rebanhos bovinos de corte, criados em uma importante área de produção pecuária, na região Nordeste do Brasil. De setembro de 2020 a agosto de 2021, foram coletadas, mensalmente, amostras fecais (n = 493) de 46 bovinos de corte. De todos os animais avaliados, larvas de nematódeos pulmonares foram detectadas em 8,7% (4/46). Os animais não apresentaram nenhum sinal clínico sugestivo de infecção por parasitos pulmonares. Vinte larvas foram recuperadas, com o número mínimo (n = 1) detectado em outubro e dezembro, e o número máximo (n = 13) em novembro. Apresentavam comprimento médio de 363 μm (± 28,65 μm), largura média de 19 μm (±1,03 μm) e eram morfologicamente semelhantes a Dictyocaulus sp.. Este estudo relata a ocorrência desse parasito nesta área. Logo, os veterinários locais devem estar atentos à inclusão desse parasito no diagnóstico diferencial de outras infecções respiratórias em bovinos de corte.

DICTYOCAULUS CERVI-LIKE LUNGWORM INFECTION IN A ROCKY MOUNTAIN ELK (CERVUS CANADENSIS NELSONI) FROM WYOMING, USA.

Dictyocaulus spp. infections are common in North American cervids, with Dictyocaulus viviparus described as most common. A Rocky Mountain elk (Cervus canadensis nelsoni) was found dead in Wyoming, US with significant bronchitis and pneumonia. In the bronchi and trachea, numerous large nematodes were found and grossly identified as Dictyocaulus spp. lungworms. Macroscopic alterations, such as distended interlobular septa and edema with foam and mucus observed on cut surface and in trachea and bronchi, were consistent with those commonly described in D. viviparus infections. Female lungworms were identified to Dictyocaulus spp. level via morphologic examination and molecular analyses based on mitochondrial cyclooxygenase 1 and 18S ribosomal RNA genes. A phylogenetic analysis was conducted employing the maximum likelihood method. Based on both morphologic and genetic assays, the isolated lungworms were most likely a strain of Dictyocaulus cervi. Within the female adult worms, free first stage larvae were observed besides worm eggs, which had not been described for Dictyocaulus spp. Phylogenetic analysis revealed that our parasites clustered closely with D. cervi, forming a subclade with that species within a larger clade that includes Dictyocaulus eckerti. While the elk tested positive for chronic wasting disease, it is assumed that significant pathology in the present case was caused directly by infection with the D. cervi-like lungworm, not previously described in North America.

Association between Dictyocaulus viviparus bulk tank milk antibody levels and farmer-reported lungworm outbreaks.

The lungworm Dictyocaulus viviparus can have a major impact on bovine health and productivity. Enzyme-Linked Immunosorbent Assays (ELISA), based on a recombinant Major Sperm Protein (MSP), have been developed to detect D. viviparus-specific antibodies in bulk tank milk (BTM). The objectives of this study are to assess the association between BTM optical density ratio's (ODR) and farmer-reported lungworm outbreaks based on the clinical sign "coughing" throughout the grazing season and to compare the sensitivity and specificity of two ELISAs under field conditions. The Hannover MSP-ELISA and the prototype Svanova MSP-ELISA were used for the detection of D. viviparus antibodies in BTM samples on 717 dairy farms during the 2018 grazing season. Assuming all herds to be truly lungworm infected, the results show that the Svanova ELISA had a lower sensitivity (40-65%) and specificity (75-90%) for the detection of D. viviparus infections in BTM compared to the Hannover ELISA, which had a sensitivity of 42% and 74% and specificity of 100% and 98% at a cut-off of 0.41 ODR and 0.25 ODR, respectively. Therefore, analyses of the associations between milk antibody levels and farmer-reported outbreaks during the 2018 and 2019 grazing season were assessed using the Hannover ELISA, on 717 and 634 farms, respectively. A positive association was found between a farmer-reported outbreak and having at least two consecutive positive BTM ODR's at a cut-off of 0.41 in 2018 (Odds Ratio (OR) = 5.5) and 2019 (OR = 2.8). Furthermore, there was a significant association between a farmer-reported outbreak and having a positive BTM ODR in August (OR 2018 = 4.4; OR 2019 = 2.8) and October (OR 2018 = 3.7; OR 2019 = 1.8). On the farms with a farmer-reported outbreak and positive BTM samples, over half (2018 = 77%; 2019 = 57%) of the positive ODR's were situated before the outbreak and 47% (2018) and 42% (2019) within 12 weeks before the outbreak. In conclusion, there is a positive association between farmer-reported outbreaks and the occurrence of a positive BTM sample at the cut-off of 0.41 ODR using the Hannover ELISA.

A survey of Dictyocaulus arnfieldi (Nematoda) infections in equids in Urmia region, Iran

Dictyocaulus arnfieldi is a lungworm commonly found in equids; however, relatively little is known about it. The aim of the present study was to establish the prevalence of equine lungworms in naturally-infected working and sporting horses, donkeys and mules in rural areas of Urmia, northwest Iran. The fecal samples were collected from 299 working horses, 57 sporting horses, 66 donkeys and 37 mules during the period March 2014 to June 2016. The collected fecal samples were processed within 48 hours following rectal sampling. The larval count/g (LPG) of feces was determined using the Baermann technique. The larvae were identified morphologically, and the arithmetic mean of the LPG was calculated at each sampling point. The overall prevalence of lungworm in all equine species was found to be 19.20%., with specific values of 15.71%, 0%, 31.81% and 24.32% being found in working horses, sporting horses, donkeys and mules, respectively. The working horses, donkeys and mules were also found to be heavily infected with Dictyocaulus arnfieldi and in addition to donkeys and mules, working horses are considered natural hosts for lungworm, and this has effects on both the epidemiology of parasitic infections and performance of the equids.

10-year parasitological examination results (2003 to 2012) of faecal samples from horses, ruminants, pigs, dogs, cats, rabbits and hedgehogs.

The results of coproscopical examinations in domestic animals and hedgehogs carried out as routine diagnostics in the years 2003 to 2012 at the Institute for Parasitology, University of Veterinary Medicine Hannover, Germany, are presented. Of 3475 horse faecal samples, 30.1% contained stages of strongyles and 1.3% eggs of Strongyloides westeri and Parascaris equorum, respectively. The most frequently observed parasite stages in 1416 cattle faecal samples were Eimeria oocysts (21.3%) and strongyle eggs or larvae (15.9%). Dictyocaulus viviparus larvae and Fasciola hepatica eggs were identified in 0.9 and 1.3% of samples. Of 574 bovine faecal samples analysed by carbol-fuchsin staining, 39.9% were positive for Cryptosporidium oocysts. Stages of strongyles were found in 52.4% of sheep (n = 374) and 44.9% of goat faeces (n = 98) and Eimeria oocysts in 41.4 and 32.7% of their faeces, respectively. Of 1848 pig faecal samples, 3.0% contained stages of strongyles, 1.6% eggs of Ascaris suum and 3.3% coccidian (Eimeria or Cystoisospora spp.) oocysts. The most frequently detected helminth eggs in faecal samples of dogs (n = 2731) and cats (n = 903) were Toxocara spp. (2.8 and 3.9%, respectively). Cystoisospora oocysts were identified in 5.6% of dog and 2.4% of cat faeces. Furthermore, 0.7% of the cat samples were positive for small Toxoplasma gondii-like oocysts. The faecal samples of rabbits (n = 434) contained eggs of Passalurus ambiguus (3.0%), strongyles (1.8%) and Trichuris leporis (0.2%) as well as Eimeria oocysts (21.2%). The most abundant nematodes in the samples of hedgehogs (n = 205) were Capillaria spp. (39.5%) and Crenosoma striatum (26.8%); coccidian oocysts were found in 14.2% of the samples.

Ectonucleotidase and adenosine deaminase as inflammatory marker in dairy cows naturally infected by Dictyocaulus viviparus.

The aim of this study was to evaluate the influence of dictyocaulosis (mild or severe) on enzymes of NTPDase, 5'-nucleotidase, and adenosine deaminase (ADA) of dairy cows naturally infected by Dictyocaulus viviparus. Blood and faeces were collected from 22 dairy cows of the same farm to evaluate NTPDase (ATP and ADP substrate), 5'-nucleotidase, and ADA activities on days 0 (pre-treatment) and 10 (post-treatment). Seric activities of NTPDase (ATP substrate), 5'-nucleotidase, and ADA were lower (P<0.05) in D. viviparus infected animals compared to uninfected cows. The number of D. viviparus larvae per gram of faeces varied among the animals, and they showed different degrees of severity according to respiratory clinical signs of the disease (cough and nasal discharge). Later, these cows were divided into two groups: those with mild (n=10) and severe (n=12) disease. Cows with severe disease showed higher NTPDase activity (ATP substrate) than those with mild disease (P≤0.05). The opposite occurred with NTPDase (ADP substrate), 5'-nucleotidase, and ADA in cows with severe disease, that is, the enzymatic activity of these seric enzymes significantly decreased (P≤0.05) compared to animals with mild disease. Infected animals showed reduced NTPDase activity (ATP and ADP substrate) after treatment. No enzymatic changes were observed for 5'-nucleotidase, and ADA pre- and post-treatment (P>0.05). Based on these results, we conclude that dictyocaulosis alters NTPDase, 5'-nucleotidase, and ADA activities of cow naturally infected by the parasite, in consequence the enzymes act as inflammatory markers.

Oxidative stress in dairy cows naturally infected with the lungworm Dictyocaulus viviparus (Nematoda: Trichostrongyloidea).

The aim of this study was to analyse the oxidative and anti-oxidant status in serum samples from dairy cows naturally infected by Dictyocaulus viviparus and its relation with pathological analyses. The diagnosis of the disease was confirmed by necropsy of one dairy cow with heavy infection by the parasite in the lungs and bronchi. Later, blood and faeces were collected from another 22 cows from the same farm to measure reactive oxygen species (ROS) levels, thiobarbituric acid-reactive substances (TBARS), catalase (CAT) and superoxide dismutase (SOD) activities on day 0 (pre-treatment) and day 10 (post-treatment with eprinomectin). Faecal examination confirmed the infection in all lactating cows. However, the number of D. viviparus larvae per gram of faeces varied between animals. Cows showed different degrees of severity according to respiratory clinical signs of the disease (cough and nasal secretion). Further, they were classified and divided into two groups: those with mild (n = 10) and severe disease (n = 12). Increased levels of TBARS (P < 0.001), ROS (P = 0.002) and SOD activity (P < 0.001), as well as reduced CAT activity (P < 0.001) were observed in cows with severe clinical signs of the disease compared to those with mild clinical signs. Eprinomectin treatment (day 10) caused a reduction of ROS levels (P = 0.006) and SOD activity (P < 0.001), and an increase of CAT activity (P = 0.05) compared to day 0 (pre-treatment). TBARS levels did not differ with treatment (P = 0.11). In summary, increased ROS production and lipid peroxidation altered CAT and SOD activities, as an adaptive response against D. viviparus infection, contributing to the occurrence of oxidative stress and severity of the disease. Treatment with eprinomectin eliminated the infection, and thus minimized oxidative stress in dairy cows.

Cattle respiratory disease in late summer and autumn.

Increased diagnoses of husk in cattle are seen as summer turns into autumn. In this article, Arthur Otter of the APHA Cattle Expert Group discusses how the disease is diagnosed, and other respiratory diseases that it might be confused with.

Genetic diversity and population genetics of large lungworms (Dictyocaulus, Nematoda) in wild deer in Hungary.

Dictyocaulus nematode worms live as parasites in the lower airways of ungulates and can cause significant disease in both wild and farmed hosts. This study represents the first population genetic analysis of large lungworms in wildlife. Specifically, we quantify genetic variation in Dictyocaulus lungworms from wild deer (red deer, fallow deer and roe deer) in Hungary, based on mitochondrial cytochrome c oxidase subunit 1 (cox1) sequence data, using population genetic and phylogenetic analyses. The studied Dictyocaulus taxa display considerable genetic diversity. At least one cryptic species and a new parasite-host relationship are revealed by our molecular study. Population genetic analyses for Dictyocaulus eckerti revealed high gene flow amongst weakly structured spatial populations that utilise the three host deer species considered here. Our results suggest that D. eckerti is a widespread generalist parasite in ungulates, with a diverse genetic backround and high evolutionary potential. In contrast, evidence of cryptic genetic structure at regional geographic scales was observed for Dictyocaulus capreolus, which infects just one host species, suggesting it is a specialist within the studied area. D. capreolus displayed lower genetic diversity overall, with only moderate gene flow compared to the closely related D. eckerti. We suggest that the differing vagility and dispersal behaviour of hosts are important contributing factors to the population structure of lungworms, and possibly other nematode parasites with single-host life cycles. Our findings are of relevance for the management of lungworms in deer farms and wild deer populations.

Development of a multiplex PCR for identification of Dictyocaulus lungworms in domestic and wild ruminants.

Dictyocaulus lungworms are the causative agents of parasitic bronchitis (dictyocaulosis) characterised by coughing and severe lung pathology in domestic and wild ruminants. The objective of this study was to design a simple molecular test that could detect of lungworm DNA from both adult and larval lungworms and could distinguish between the most common Dictyocaulus species found in cattle and in some species of wild ruminants. A multiplex PCR test with four novel primers targeting species-specific regions of the second internal transcribed spacer (ITS2) was designed based on our own sequence data as well as on available sequence information in GenBank. After PCR amplification of lungworms from European bison (Bison bonasus), cattle (Bos taurus), moose (Alces alces), red deer (Cervus elaphus) and roe deer (Capreolus capreolus), products were analysed with gel electrophoresis. This resulted in three specific bands of different size depending on the species analysed. Dictyocaulus viviparus collected from cattle or European bison resulted in a ca. 560 bp band, D. capreolus collected from roe deer produced a band ca. 400 bp and the longest DNA band (ca. 660 bp) was obtained with DNA from Dictyocaulus sp. collected from red deer and moose. Dictyocaulus eckerti bands with expected size of 714 bp were not observed in our study. The multiplex method produced consistent results with samples from both Sweden and Poland and overcame the limitations of traditional techniques based on differences in morphological features of parasites at different life stages.

Association between Dictyocaulus viviparus status and milk production parameters in Dutch dairy herds.

The objective of this study was to estimate the association between Dictyocaulus viviparus bulk tank milk (BTM) test results and milk production and milk composition parameters in adult Dutch dairy cattle herds. Bulk tank milk samples were collected in August and November 2013, and ELISA tests were performed. Two hundred BTM positive (BTM+) and 200 BTM negative (BTM-) herds were selected based on their BTM test result of November 2013, obtained from a list of farms that participated in the Dutch GD Animal Health voluntary monitoring program for controlling nematode infections. The relationship between D. viviparus BTM status and 3 production parameters (milk production, milk fat %, and milk protein %) in summer (June to August 2013) and autumn (September and October 2013) was investigated using generalized linear mixed models. Production data were available for 126 BTM- herds and 109 BTM+ herds. Results showed that a positive D. viviparus status was associated with decreased milk production (June: -1.01, July: -1.19, August: -1.68, September and October: -1.33kg/cow per d). Milk fat percentage was 0.14% and 0.08% lower during summer and autumn, respectively, in BTM+ herds. No significant association was demonstrated between a positive BTM test result and milk protein percentage. Because a strong correlation was present between the BTM status for D. viviparus and that for Ostertagia ostertagi, these losses cannot be attributed to one of the two parasites. However, it is clear that these parasite infections have a considerable effect on production.

Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle.

BACKGROUND: A major constraint for the effective control and management of helminth parasites is the lack of rapid, high-throughput, routine diagnostic tests to assess the health status of individual animals and herds and to identify the parasite species responsible for these helminthoses. The capability of a multiplex platform for the simultaneous detection of three pasture associated parasite species was evaluated and compared to existing ELISAs. METHODS: The recombinant antigens 14.2 kDa ES protein for Cooperia oncophora, major sperm protein for Dictyocaulus viviparus and Cathepsin L1 for Fasciola hepatica were recombinantly expressed either in Escherichia coli or Pichia pastoris. Antigens were covalently coupled onto magnetic beads. Optimal concentrations for coupling were determined following the examination of serum samples collected from experimentally mono-infected animals, before and after their infection with the target species. Absence of cross-reactivity was further determined with sera from calves mono-infected with Haemonchus contortus, Ostertagia ostertagi and Trichostrongylus colubriformis. Examination of negative serum samples was characterised by low median fluorescence intensity (MFI). RESULTS: Establishment of the optimal serum dilution of 1:200 was achieved for all three bead sets. Receiver Operating Characteristic analyses were performed to obtain cut-off MFI values for each parasite separately. Sensitivity and specificity at the chosen cut-off values were close to, or 100% for all bead sets. Examination of serum samples collected on different days post infection from different animals showed a high reproducibility of the assays. Serum samples were additionally examined with two already established ELISAs, an in-house ELISA using the recombinant MSP as an antigen and a DRG ELISA using Cathepsin L1 for liver fluke. The results between the assays were compared and kappa tests revealed an overall good agreement. CONCLUSIONS: A versatile bead-based assay using fluorescence detection (xMAP technology) was developed to simultaneously detect antibodies against C. oncophora, D. viviparus and F. hepatica in cattle serum samples. This platform provides rapid, high-throughput results and is highly sensitive and specific in comparison to existing serological as well as coproscopical diagnostic techniques.

Cytokine mRNA expression in bronchoalveolar lavage cells during Dictyocaulus viviparus infection in calves.

The purpose of this study was to monitor local cytokine responses to Dictyocaulus viviparus in calves during primary infection and re-infection. Bronchoalveolar lavage fluid (BALF) was collected weekly from experimentally infected calves and interleukin-2 (IL-2), IL-4, IL-5, IL-10 and IFN-γ mRNA expression was quantified in BALF cells. The major finding was a prominent transient increase in IL-4 mRNA expression, compared with that of uninfected calves, observed in BALF cells collected 2-3 weeks post-primary D. viviparus infection. At 2 weeks post-infection, macroscopic worms were also first observed in BALF. Calves re-infected after 10 weeks were partially immune which was evident at slaughter 5 weeks post-infection as a lower worm burden than in previously naïve calves infected at the same time. IL-4 mRNA expression in BALF cells 2 weeks post-re-infection was increased compared with that of uninfected animals but not as high as that of primarily infected calves. BALF cell expression of the other cytokines tested for was not as clearly effected by the D. viviparus infection. It seems likely that the strong IL-4 response observed during primary infection reflects an innate response to the worms that may initiate an ensuing Th2 response, which confers protective immunity.

Lungworm Infections in German dairy cattle herds--seroprevalence and GIS-supported risk factor analysis.

In November 2008, a total of 19,910 bulk tank milk (BTM) samples were obtained from dairy farms from all over Germany, corresponding to about 20% of all German dairy herds, and analysed for antibodies against the bovine lungworm Dictyocaulus viviparus by use of the recombinant MSP-ELISA. A total number of 3,397 (17.1%; n = 19,910) BTM samples tested seropositive. The prevalences in individual German federal states varied between 0.0% and 31.2% positive herds. A geospatial map was drawn to show the distribution of seropositive and seronegative herds per postal code area. ELISA results were further analysed for associations with land-use and climate data. Bivariate statistical analysis was used to identify potential spatial risk factors for dictyocaulosis. Statistically significant positive associations were found between lungworm seropositive herds and the proportion of water bodies and grassed area per postal code area. Variables that showed a statistically significant association with a positive BTM test were included in a logistic regression model, which was further refined by controlled stepwise selection of variables. The low Pseudo R(2) values (0.08 for the full model and 0.06 for the final model) and further evaluation of the model by ROC analysis indicate that additional, unrecorded factors (e.g. management factors) or random effects may substantially contribute to lungworm infections in dairy cows. Veterinarians should include lungworms in the differential diagnosis of respiratory disease in dairy cattle, particularly those at pasture. Monitoring of herds through BTM screening for antibodies can help farmers and veterinarians plan and implement appropriate control measures.

Study on small ruminant lungworms and associated risk factors in northeastern Iran.

OBJECTIVE: To determine the prevalence, identify the species involved and assess possible risk factors of lungworm infection in small ruminant slaughtered in abattoir of Mashhad in the northeast of Iran from October 2010-August 2011. METHODS: Faecal and post mortem examination were conducted on 350 and 2 500 animals, respectively. RESULTS: The overall prevalence of lungworm infection was 10.85% and 3.80% in coproscopic and post mortem examination respectively, and this difference was found to be significant. Higher prevalence of lungworm infection was recorded in post mortem examination in sheep (4.1%) than in goats (0.5%) (P< 0.05). The proportion of infection with Dictyocaulus filaria, Protostrongylus rufescens and mixed infection were 3.7%, 0.1% and 0.2% in sheep while in goats, the infection was reported with Dictyocaulus filaria (0.5%) only. The seasonal dynamics of lungworm infection showed that prevalence was highest in winter (7.8%) with a remarkable decline during the dry time (summer) (0.8%) which the difference was significant (P<0.001). The animals of less than one year old showed greater infection in post mortem examination than older animals significantly (P<0.001). Also, the infection rate between male and female animals showed significant difference (P<0.05) with prevalence rate of 4.6% and 2.0%, respectively. CONCLUSIONS: Due to its impact on production, emphasis should be given for the control and prevention of lungworm infection in this area.

Assessment of the genetic relationship between Dictyocaulus species from Bos taurus and Cervus elaphus using complete mitochondrial genomic datasets.

BACKGROUND: Dictyocaulus species are strongylid nematodes of major veterinary significance in ruminants, such as cattle and cervids, and cause serious bronchitis or pneumonia (dictyocaulosis or "husk"). There has been ongoing controversy surrounding the validity of some Dictyocaulus species and their host specificity. Here, we sequenced and characterized the mitochondrial (mt) genomes of Dictyocaulus viviparus (from Bos taurus) with Dictyocaulus sp. cf. eckerti from red deer (Cervus elaphus), used mt datasets to assess the genetic relationship between these and related parasites, and predicted markers for future population genetic or molecular epidemiological studies. METHODS: The mt genomes were amplified from single adult males of D. viviparus and Dictyocaulus sp. cf. eckerti (from red deer) by long-PCR, sequenced using 454-technology and annotated using bioinformatic tools. Amino acid sequences inferred from individual genes of each of the two mt genomes were compared, concatenated and subjected to phylogenetic analysis using Bayesian inference (BI), also employing data for other strongylids for comparative purposes. RESULTS: The circular mt genomes were 13,310 bp (D. viviparus) and 13,296 bp (Dictyocaulus sp. cf. eckerti) in size, and each contained 12 protein-encoding, 22 transfer RNA and 2 ribosomal RNA genes, consistent with other strongylid nematodes sequenced to date. Sliding window analysis identified genes with high or low levels of nucleotide diversity between the mt genomes. At the predicted mt proteomic level, there was an overall sequence difference of 34.5% between D. viviparus and Dictyocaulus sp. cf. eckerti, and amino acid sequence variation within each species was usually much lower than differences between species. Phylogenetic analysis of the concatenated amino acid sequence data for all 12 mt proteins showed that both D. viviparus and Dictyocaulus sp. cf. eckerti were closely related, and grouped to the exclusion of selected members of the superfamilies Metastrongyloidea, Trichostrongyloidea, Ancylostomatoidea and Strongyloidea. CONCLUSIONS: Consistent with previous findings for nuclear ribosomal DNA sequence data, the present analyses indicate that Dictyocaulus sp. cf. eckerti (red deer) and D. viviparus are separate species. Barcodes in the two mt genomes and proteomes should serve as markers for future studies of the population genetics and/or epidemiology of these and related species of Dictyocaulus.

A survey on Dictyocaulus viviparus antibodies in bulk milk of dairy herds in Northern Germany.

Parasitic bronchitis caused by the bovine lungworm, Dictyocaulus viviparus, occurs worldwide in temperate areas. The parasite is found predominantly in calves and heifers, but dairy cattle can suffer from lungworms when they become infected for the first time or if they have lost immunity due to lack of exposure to lungworm larvae during the grazing season. The present study was performed to determine the D. viviparus bulk milk antibody prevalence in dairy herds in the East Frisian region of northwestern Germany, Lower Saxony, by analysing bulk milk samples collected in January (860 samples), September (866 samples) and November (860 samples) 2008, thereby representing 906 dairy farms. These samples were tested for antibodies against D. viviparus by a milk ELISA. This test detects patent infections only since it is based on recombinant major sperm protein as antigen. While in January 12.8% of dairy farms were positive for D. viviparus antibodies, the bulk milk samples collected in September and November revealed 6.9% and 6.6% positive dairy herds. From the 906 dairy farms included in the study, 191 (21.1%) tested positive at least once for antibodies against lungworm. From 810 dairy farms from which bulk milk samples were obtained during all three samplings, 146 (18.0%) farms were positive at one sampling date, 27 (3.3%) at two, and 4 (0.5%) on all three sampling dates. The majority of the farms represented in the study belonged to four districts of East Frisia, which showed no significant difference in the proportion of positive dairy farms.

Efficacy of eprinomectin pour-on against Dictyocaulus arnfieldi infection in donkeys (Equus asinus).

A trial to assess the efficacy of eprinomectin (EPM) against the lungworm Dictyocaulus arnfieldi was carried out on 15, naturally-infected donkeys. Ten animals were treated with a 'pour-on' EPM preparation (at a dose of 0.5 mg/kg bodyweight), and five animals acted as controls. Faecal larval counts were carried out two days before treatment, on day of treatment and 7, 14, 21 and 28 days post-treatment with the anthelmintic. EPM was 100% effective in eliminating faecal larvae from day 7, until the end of study at day 28. No adverse drug-reactions or side-effects were observed in any of the treated donkeys.