Predicting the unpredictable? A climate-based model of the timing of peak pasture infectivity for Dictyocaulus viviparus.

Outbreaks of cattle lungworm disease (Dictyocaulus viviparus) are explosive and costly. The unpredictability of the disease often encourages farmers to apply blanket anthelmintic treatments to the herd, which impede the acquisition of immunity, increase the risk of drug resistance, and interfere with efforts to reduce anthelmintic use against ubiquitous gastrointestinal nematodes. Improving our understanding of the factors which lead to a high risk of infection with lungworm, (including climatic pressure), would support a more targeted management. We present GLOWORM-FL-DV, the first mathematical model of the free-living stages of D. viviparus. The ecology of D. viviparus is unique compared with other strongylid nematodes due to its relationship with Pilobilus spp. fungi, which enhance the transmission potential. The role of the fungi was therefore incorporated into the model framework, informed by laboratory observations of Pilobolus spp. development and sporulation. The thermal niche of D. viviparus was characterised based on published and laboratory observations. Mortality of parasitic larvae increased significantly below 0oC, and larval development occurred above 1.4oC, whereas the estimated minimum temperature for migration via Pilobolus spp. was 8.8oC. Model predictions were compared with antibody levels in bulk milk tank samples collected at two-weekly intervals from eight dairy herds across Great Britain over two grazing seasons. The model predicted high levels of larval abundance on pasture 46 days (38-52 days) before a rise in antibody levels and 22-26 days before the onset of clinical signs. The model assesses the impact of climate and weather on lungworm larval availability at pasture and provides a framework for the development of a risk forecasting system. This could help to focus vigilance for clinical signs at high-risk times and facilitate the targeted use of anthelmintics to prevent outbreaks, in support of sustainable parasite control.

Gastrointestinal nematode and lungworm infections in organic dairy calves reared with nurse cows during their first grazing season in western France.

The rearing system of dairy calves with nurse cows has been developing since 2010 in organic farms in western France. This system allows cow-calf contact until a weaning age close to the natural weaning for cattle and is characterized by an early turnout for calves at around one month of age with their nurse cows and a first grazing season with mixed grazing of calves and adults at a ratio of 2-4 calves per nurse cow. The objectives of this study were to assess the gastrointestinal (GIN) and lungworm infections in such reared calves and their variability during the first grazing season. Faecal egg count (FEC), pepsinogen (PEP) concentration and Ostertagia ELISA optical density ratio (ODR) were determined in calves (n = 497) at housing in 33 groups from 24 farms in 2018, and in calves (n = 405) and nurse cows (n = 199) throughout the 2019 grazing season in 41 groups from 20 farms. For lungworm infection, information was obtained during 2019 through the recording of coughing episodes along the grazing season and the Dictyocaulus ELISA ODR determination at housing both in calves and nurses. Results indicated that the level of GIN infection was overall low for calves during the first grazing season with PEP and Ostertagia ODR group-average values ranging from 0.97 to 1.6 U Tyr and 0.23 to 0.71 ODR respectively. No anthelmintic treatment being given in any group of calves. Ostertagia ODR values increased with the duration of the grazing season (>240 d) and with the ratio calves/nurse (>2). GIN parameters for nurses remained fairly stable during the grazing season with mean FEC, PEP and Ostertagia ODR group-average values of 13 epg, 2.28 U Tyr and 0.81 ODR, respectively. Antibodies against lungworms were detected in 3-62 % of calves depending on the duration of grazing, but only 6% of calves showed a coughing episode. The dilution effect due to the mixed grazing of resistant (nurse cows) and susceptible (calves) animals associated with predominant milk diet of calves during the first months of grazing in combination with protective grazing management allow calves to be turned out at an early age without using anthelmintic treatments. Further studies are needed to assess the GIN infection dynamics during the second grazing season in weaned heifers.

Immunization Trials with Recombinant Major Sperm Protein of the Bovine Lungworm Dictyocaulus viviparus.

The lungworm Dictyocaulus viviparus is one of the most economically important bovine parasites in temperate climate regions. Following infection, D. viviparus induces a temporary protective immunity, and a vaccine based on attenuated, infective larvae is commercially available. However, due to several disadvantages of the live vaccine, the development of a recombinant subunit vaccine is highly desirable. Therefore, the major sperm protein (MSP), which is essential for the parasite's reproduction, was tested as a recombinantly Escherichia coli-expressed glutathione-S-transferase (GST)-fused vaccine antigen in immunization trials with two different adjuvants, Quil A and Al(OH)3. Calves (N = 4 per group) were immunized on study day (SD) 0, 21 and 42 and given a challenge infection on SD 63-65. The two control groups received only the respective adjuvant. Based on geometric means (GM), a 53.64% reduction in larvae per female worm was observed in the rMSP Quil A group vs. its control group (arithmetic means (AM): 54.43%), but this difference was not statistically significant. In the rMSP Al(OH)3 group, the mean number of larvae per female worm was even higher than in the respective control group (GM: 9.24%, AM: 14.14%). Furthermore, male and female worm burdens and the absolute number of larvae did not differ significantly, while the Al(OH)3 control group harbored significantly longer worms than the vaccinated group. Vaccinated animals showed a rise in rMSP-specific antibodies, particularly IgG and its subclass IgG1, and the native protein was detected by immunoblots. Although rMSP alone did not lead to significantly reduced worm fecundity, it might still prove useful as part of a multi-component vaccine.

Seroprevalence of Major Pasture-Borne Parasitoses (Gastrointestinal Nematodes, Liver Flukes and Lungworms) in German Dairy Cattle Herds, Association with Management Factors and Impact on Production Parameters.

Pasture-borne parasites adversely affect bovine health and productivity worldwide. In Europe, gastrointestinal nematodes, especially Ostertagia ostertagi, the liver fluke Fasciola hepatica and the lungworm Dictyocaulus viviparus represent the most important parasites of dairy cattle. The present study assessed exposure towards these parasites among 646 cattle herds in three parts of Germany during 2017-2019 via antibody detection in bulk tank milk (BTM). Overall, O. ostertagi levels indicative of production losses were detected in 41.2% (266/646; 95% confidence interval (CI): 37.4-45.1%) of BTM samples, while F. hepatica seroprevalence amounted to 14.9% (96/646; 95% CI: 12.2-17.9%). Only 2.3% (15/646; 95% CI: 1.4-3.9%) of samples were D. viviparus antibody-positive. Significantly lower O. ostertagi as well as F. hepatica seroprevalence was detected in dual-purpose breeds compared to high-performance breeds from the same region. Management factors related to parasite exposure included access to fresh grass and hay, silage quality and anthelmintic treatment. Furthermore, F. hepatica and O. ostertagi seropositivity was significantly associated with suboptimal herd-level body condition. Interestingly, the relationship between seropositivity and productivity differed between breed types. Negative impacts on milk yield were detected only in high-performance breeds, while O. ostertagi seropositivity was associated with a lower milk fat content in dual-purpose herds.

Large lungworms (Nematoda: Dictyocaulidae) recovered from the European bison may represent a new nematode subspecies.

Although the Dictyocaulus lungworm, the agent of dictyocaulosis, is one of parasitological threats to European bison, its systematic position remains unclear. The aim of the present study was to evaluate the morphological features of the lungworm and the pathological lesions it induces, and to analyse mitochondrial (mt) genetic markers for systematic and molecular epidemiological studies. The morphological findings indicate that Dictyocaulus lungworms of European bison can be distinguished from those of cattle on the basis of differences in buccal capsule wall length, total body length, and spicules length in males, all of which were significantly longer in those of European bison. Nucleotide diversity calculated from pairwise sequence alignments of partial cytochrome c oxidase subunit 1 (cox1), cytochrome B (cytB) and NADH dehydrogenase subunit 5 (nad5) of specimens from cattle and European bison varied from 1.7% for nad5, 2.1% for cytB, to 3.7% for cox1 gene. Thus, among the lungworms of European bison and cattle, nad5 and cytB were the most conserved proteins, whereas cox1 was the most diverse. The mt cytB marker gene may be a suitable candidate for distinguishing between the two genotypes, as nad5 demonstrated the greatest within-genus sequence variation. The lung tissue of infected European bison manifests signs of verminous pneumonia characterized by interstitial pneumonia, bronchitis and bronchiolitis. Therefore, it appears that European bison and cattle are infected with slightly diverged, morphologically-different, genotypes of D. viviparus, indicating they belong to two separate worm populations. We propose, therefore, that the lungworm of European bison should be classified as D. viviparus subsp. bisontis.

A novel pooled milk test strategy for the herd level diagnosis of Dictyocaulus viviparus.

Current diagnostic methods for detecting the presence or absence of Dictyocaulus viviparus in dairy herds, are insensitive when based on detection of antibody levels in bulk tank milk (BTM). Here we present a novel technique to confirm the presence of the parasite based on a pooled-milk sample from 10 randomly selected first - lactation heifers (FLH). This study was run in two parts. First, a longitudinal study was performed to look at infection dynamics in milk samples across the grazing season using a prototype ELISA developed by Svanova (Boehringer-Ingelheim, Uppsala). We identified that mean ODR values in milk samples from FLH was significantly higher than that for older cows (0.13 versus 0.07 respectively, p < 0.001) suggesting that samples from the FLH cohort should be pooled to produce the test. Second, the pooled - milk test was evaluated on a cross-sectional survey of UK dairy herds (n = 25 grazing and n = 25 zero-grazing herds) to evaluate test performance under field conditions. The optical density ratio (ODR) cut-off value for our pooled-milk test using 10 FLH milk samples was optimal at a value of 0.16. Pooling 10 FLH samples created a sensitivity and specificity of 66.7% and 95.5% respectively. In comparison, whole-herd BTM samples had a maximum sensitivity of 37.5% and specificity of 63.6% at an ODR cut-off of 0.18. The area under the curve according to receiver-operative-characteristic (ROC) analysis was high for the 10-heifer test (0.87) but poor for the whole herd BTM testing (0.45). This study provides a more sensitive diagnostic test strategy for the screening of D.viviparus in dairy herds. Testing herds at the end of a grazing season would facilitate the planning of effective control measures, such as the use of the lungworm vaccination or strategic deworming, for the following grazing season. This may prove to be a useful test strategy for the diagnosis of a variety of parasitic diseases of livestock.

A novel pooled milk test strategy for the herd level diagnosis of Dictyocaulus viviparus.

Current diagnostic methods for detecting the presence or absence of Dictyocaulus viviparus in dairy herds, are insensitive when based on detection of antibody levels in bulk tank milk (BTM). Here we present a novel technique to confirm the presence of the parasite based on a pooled-milk sample from 10 randomly selected first - lactation heifers (FLH). This study was run in two parts. First, a longitudinal study was performed to look at infection dynamics in milk samples across the grazing season using a prototype ELISA developed by Svanova (Boehringer-Ingelheim, Uppsala). We identified that mean ODR values in milk samples from FLH was significantly higher than that for older cows (0.13 versus 0.07 respectively, p < 0.001) suggesting that samples from the FLH cohort should be pooled to produce the test. Second, the pooled - milk test was evaluated on a cross-sectional survey of UK dairy herds (n = 25 grazing and n = 25 zero-grazing herds) to evaluate test performance under field conditions. The optical density ratio (ODR) cut-off value for our pooled-milk test using 10 FLH milk samples was optimal at a value of 0.16. Pooling 10 FLH samples created a sensitivity and specificity of 66.7% and 95.5% respectively. In comparison, whole-herd BTM samples had a maximum sensitivity of 37.5% and specificity of 63.6% at an ODR cut-off of 0.18. The area under the curve according to receiver-operative-characteristic (ROC) analysis was high for the 10-heifer test (0.87) but poor for the whole herd BTM testing (0.45). This study provides a more sensitive diagnostic test strategy for the screening of D.viviparus in dairy herds. Testing herds at the end of a grazing season would facilitate the planning of effective control measures, such as the use of the lungworm vaccination or strategic deworming, for the following grazing season. This may prove to be a useful test strategy for the diagnosis of a variety of parasitic diseases of livestock.

Gastrointestinal and pulmonary nematodes in calves naturally infected in the cities of Botucatu and Manduri, in the Brazilian state of São Paulo / Nematódeos gastrintestinais e pulmonares em bezerros naturalmente infectados nos municípios de Botucatu e Manduri, estado de São Paulo

This study aimed to determine the occurrence of gastrointestinal and pulmonary parasites in calves and to evaluate seasonal and age patterns in parasitism. For this, we used 140 clinically healthy crossbreed calves (two to 12 months old) that belonged to two private farms in the municipalities of Botucatu (n=53) and Manduri (n=87), São Paulo state, Brazil. The calves were monitored for 12 months (from September 2014 to August 2015). Fecal samples were collected directly from the rectum every three months. Fecal egg counts were determined using the modified McMaster technique with a sensitivity of 50 eggs per gram of feces (EPG). Coproculture was performed on pooled samples to identify Strongylida infective larvae. First-stage larvae of Dictyocaulus viviparus were extracted by a modified Baermann method. The data showed non-normal distribution (Shapiro-Wilk) and the nonparametric Kruskall-Wallis test was employed to evaluate the EPG data by seasons and age groups. Dunn's post-test was used for multiple comparisons (P<0.05). The calves from Manduri farm showed significantly higher fecal egg counts (P<0.0001) in the winter when compared to other seasons. At Botucatu farm, young calves (2-3 months old) showed significantly higher EPG than old calves (8-12 months) (P=0.01). The prevalence and overall mean of animals positive for Strongylida type-eggs were 81.1% and 340 in Botucatu, respectively, versus 83.9% and 854 in Manduri, respectively. Furthermore, we found Strongyloides spp., Moniezia spp., and Trichuris spp. eggs and Eimeria spp. oocysts. The prevalent genera in all coprocultures in decreasing order were: Cooperia spp., Haemonchus spp., Oesophagostomum spp., and Trichostrongylus spp. First-stage larvae of Dictyocaulus viviparus were found only in Botucatu farm samples throughout the year, except in spring.(AU)

O objetivo deste estudo foi investigar os parasitas gastrintestinais e pulmonares que acometem bezerros bem como a possível influência de fatores climáticos e da idade no parasitismo. Para isso, durante um período de 12 meses (setembro de 2014 a agosto de 2015), amostras de fezes foram coletadas a cada três meses diretamente da ampola retal de 140 bezerros mestiços (dois a 12 meses de idade), clinicamente saudáveis, pertencentes a duas propriedades leiteiras localizadas nos municípios de Botucatu (n=53) e Manduri (n=87), estado de São Paulo. Realizou-se a contagem de ovos por grama de fezes pela técnica de McMaster modificada com sensibilidade de 50 ovos por grama de fezes (OPG). Coproculturas foram realizadas em pool de amostras para a obtenção das larvas infectantes (L3). Larvas de primeiro estágio de Dictyocaulus viviparus foram recuperadas pela modificação da técnica de Baermann. Os dados não se apresentaram normalmente distribuídos (Shapiro-Wilk), e o teste não paramétrico de Kruskal-Wallis foi utilizado para avaliar os dados de OPG em relação às estações do ano e faixa etária. Para comparações múltiplas, empregou-se o pós-teste de Dunn. Foi verificado que em Manduri, no inverno, houve um aumento significativo (P<0,0001) na contagem de OPG em comparação as demais estações do ano. Em Botucatu, os animais com dois a três meses de idade apresentaram maiores contagens de OPG quando comparados aos animais de oito a 12 meses de idade (P=0,01). A prevalência e a média global de animais positivos para ovos do tipo Strongylida, em Botucatu, foi de 81,1% e 340, respectivamente, e em Manduri foi de 83,9% e 854, respectivamente. Em adição, de maneira geral, foram encontrados ovos de Strongyloides spp., Moniezia spp., Trichuris spp. e oocistos de Eimeria spp. Foram recuperadas, em ordem de prevalência, larvas infectantes de Cooperia spp., Haemonchus spp., Oesophagostomum spp. e Trichostrongylus spp. Larvas de D. viviparus foram recuperadas somente na propriedade de Botucatu durante todo o ano, com exceção da primavera.(AU)

Apparent prevalence of and risk factors for infection with Ostertagia ostertagi, Fasciola hepatica and Dictyocaulus viviparus in Swiss dairy herds.

Infections with helminth parasites can negatively affect performance of dairy cows. Knowledge on infection intensity, spatial distributions and risk factors are key to develop targeted treatment strategies. Canada and most EU countries have conducted large investigations, but respective data for Switzerland were missing. We now performed a bulk tank milk serosurvey for Ostertagia ostertagi, Fasciola hepatica, and Dictyocaulus viviparus on a total of 1036 voluntarily participating dairy herds that were sampled at confinement periods, i.e. in winter 2014/15 or 2015/16, respectively. All samples were analyzed with commercial ELISAs for antibodies (AB) against O. ostertagi and F. hepatica, and those of the first sampling period additionally with an in-house ELISA for AB against D. viviparus. Testing for the latter parasite was not done in the second year of the study, as the sampling period might have missed infections due to the short lived nature of specific antibodies. The possible influence of geographic, climatic, and farm management variables on AB levels were assessed for each parasite using scanning cluster and multiple regression analysis. Overall seroprevalence for O. ostertagi was 95.5% (95% C.I.: 94.0-96.6), with a mean optical density ratio (ODR) of 0.83, for F. hepatica 41.3% (95% C.I.: 38.3-44.4), and for D. viviparus 2.9% (95% C.I.: 1.6-4.7). There were no significant differences between the two sampling periods. For all parasites, significant geographic clusters of higher AB levels could be established. Furthermore, AB levels against all three parasites were positively correlated with each other, indicating either cross-reactions or co-infections. For O. ostertagi, herd size and percentage of pasture in the ration were positively correlated with AB levels. For F. hepatica, altitude above sea level (a.s.l.) positively, and milk production per cow and year was negatively correlated with AB levels. This work provides baseline data for further studies performing in-depth risk factor analysis and investigating management as well as targeted treatment options to control the parasites.

The effect of patent Dictyocaulus viviparus (re)infections on individual milk yield and milk quality in pastured dairy cows and correlation with clinical signs.

BACKGROUND: Infections with the bovine lungworm Dictyocaulus viviparus might lead to reduced milk production and detrimental impacts on milk quality resulting in considerable economic losses in dairy farming. METHODS: In the presented field study, 1988 faecal samples were collected from 1166 Black and White dairy cows allocated in 17 small and medium-sized German grassland farms. Faecal samples were collected in summer and autumn 2015 to assess D. viviparus larvae excretion. Test-day records were used to estimate the association between patent D. viviparus infections in individual cows and the milk production parameters milk yield, milk protein and milk fat content by using linear mixed models. Bulk tank milk (BTM) samples from each farm and individual milk samples from those cows which were excreting larvae in summer were collected in autumn. In addition, occurrence of the clinical symptom "coughing" was noted in individual cows during autumn sampling to determine its association with patent lungworm infections. RESULTS: Patent D. viviparus infections were found on 23.5% (4/17) of farms with a prevalence at the individual cow level of 0.9% (9/960) in summer and 3.4% (35/1028) in autumn. No BTM sample exceeded the BTM ELISA cut-off value of 0.410 optical density ratio (ODR), the mean value was 0.168 ODR. Only one individual milk sample exceeded the individual milk ELISA cut-off value of 0.573 ODR (mean value of 0.302 ODR). A patent D. viviparus infection status was associated with a lower average daily milk yield of 1.62 kg/cow/day (P = 0.0406). No significant association was found with milk protein or fat content representing milk quality parameters. Coughing was observed in 5.9% (61/1028) of cows. Of the coughing cows, only 4.9% (3/61) had a patent lungworm infection. Fisher's exact test showed no significant difference between infected and non-infected coughing cows. CONCLUSIONS: Farmers and veterinarians should be aware that patent lungworm (re)infections in dairy cows reduce milk yield, despite the absence of clinical signs. Furthermore, if dairy cows present with coughing, other differential diagnoses need to be considered in addition to dictyocaulosis.

Environmental factors associated with Dictyocaulus viviparus and Fasciola hepatica prevalence in dairy herds from Costa Rica.

Dictyocaulosis and fasciolosis are parasitic diseases that cause considerable economic losses for owners of farm animals worldwide, with special relevance on fasciolosis because it is an emerging zoonosis. Indirect diagnosis of these diseases through analyses of bulk milk tank samples has allowed carrying out large-scale prevalence studies, while the use of geographical information systems has helped to visualize and determine those variables that affect distribution of these pathogens. This study is intended to describe the spatial distribution of Dictyocaulus viviparus and Fasciola hepatica in dairy herds from Costa Rica, as well as their associated environmental factors. Bulk milk tank samples from 526 dairy herds in the three most important dairy regions of Costa Rica were analyzed using enzyme immunoassays. Results from the farms were subjected to spatial analyses using Holdridge's life zones, relief and soil type environmental layers. Of the total bulk milk tank samples analyzed, 3.8% (n=20) and 3.6% (n=19) were positive for D. viviparus and F. hepatica, respectively. Moran's I analysis revealed the existence of potential cluster (Moran's I=1.789, z=12.726 p<0.05) for D. viviparus. Consequently, Getis-Ord General G analysis showed that the spatial distribution of positive farms in the dataset was clustered (Observed General G=0.015, variance=0.000001, z=12.823, p<0.05). No significant positive spatial autocorrelation (Moran's I=0.038, z=0.286, p>0.0.5) was observed for F. hepatica. Furthermore, a significant difference was detected in the spatial locations of both parasites (latitude p<0.05, longitude p<0.05), and about the spatial distribution of both D. viviparus negative and positive farms (latitude p<0.05, longitude p<0.05), as well as in F. hepatica negative and positive farms regarding on latitude (p<0.05), but not on longitude (p>0.05). In the case of environmental factors, significant differences were found for D. viviparus and F. hepatica with respect to types of soil, precipitation, altitudinal belts, life zones, biotemperature, and elevation.

Changes in biochemical analytes in calves infected by nematode parasites in field conditions.

Parasitic infections caused by nematodes are a major problem in bovines that resulting in losses in animal health and production. Thus, the aim of this study was to evaluate alterations in selected serum biochemical analytes in calves naturally infected with gastrointestinal (GI) and pulmonary nematodes without clinical signs. For this, samples of feces and blood of 86 calves were collected. Fecal egg counts (FEC) were determined using the modified McMaster technique with a sensitivity of 50 eggs per gram of feces (EPG). Positive nematode FEC was processed for coproculture using pooled samples to identify Strongylidae infective larvae (L3). First stage-larvae (L1) of Dictyocaulus viviparous were identified by a modified Baermann method. The biochemical analytes determined were: acute phase proteins such as haptoglobin and paraoxonase type 1; the enzymes acetylcholinesterase; butyrylcholinesterase; the lipid profile (triglycerides and total, HDL, and LDL-cholesterol); serum iron profile (iron and unsaturated iron-binding capacity); total protein and albumin; pancreatic profile (amylase and lipase); and minerals (phosphorus and calcium). The calves were divided into four groups according to the results of EPG and the modified Baermann method. Group 1: healthy control animals (n=16); Group 2: calves with only GI parasites (n=51): This group was sub-divided into sub-groups according to the EPG threshold: 2a-GI parasites with low EPG (n=23), and 2b-GI parasites with high EPG (n=28). Group 3: animals with only lungworms (n=5), and Group 4: calves with lung+GI parasites (n=14). The more prevalent genera in all coprocultures were: Cooperia spp., Haemonchus spp., Oesophagostomum spp., and Trichostrongylus spp. The nonparametric Kruskal-Wallis test was used to compare the groups and Dunn's post-test was used for multiple comparisons as the data was not normally distributed (P<0.05). The haptoglobin concentration increased in calves with GI and pulmonary parasites. A significant increase in acetylcholinesterase was observed in calves infected with lungworms. Cholesterol, triglycerides, HDL, and LDL concentrations decreased but lipase concentration increased in calves with GI parasites. Therefore, this paper provides an overview of the biochemical effects produced by nematode parasites in calves in field conditions. These findings in calves without any evident clinical signs of disease could provide an indication of GI parasites and lungworm infection, especially in an endemic area for these parasites.

Vaccination of calves with yeast- and bacterial-expressed paramyosin from the bovine lungworm Dictyocaulus viviparus.

Previously, vaccination of cattle with Escherichia coli-expressed bovine lungworm paramyosin (EcPMY) adjuvanted with Quil A resulted in considerable reduction in worm burden and larvae shedding (Strube et al., 2015). To further evaluate the protective potential of PMY, cattle vaccination trials were performed using either E. coli- (EcPMY) or Pichia pastoris-expressed PMY (PpPMY) with different adjuvants (Matrix-Q(™) or Quil A). Combinations EcPMY+Matrix-Q(™) (trial 1), PpPMY+Matrix-Q(™) (trial 2) and PpPMY+Quil A (trial 3) were tested against challenge infections with 2000 Dictyocaulus viviparus larvae. Even though GM worm burden and larvae shedding was lower in almost all vaccinated groups, there were high variations between individuals hampering significant differences. However, in all vaccinated groups, lungworms were significantly shorter compared with those in controls. In vitro stimulation of peripheral blood mononuclear cells (PBMC) with recombinant (r)PMY revealed no significant proliferation following vaccinations or challenge infection. All vaccinated cattle showed a significant rise in specific antibodies, particularly IgG and its subclass IgG1, and detected the native lungworm PMY in immunoblots starting 2 weeks after the first vaccination. The use of a different rPMY-adjuvant combination or combined vaccination with additional recombinant antigens might be a promising future approach towards a new vaccine against lungworms in cattle.

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.

Prevalence and seasonality of bulk milk antibodies against Dictyocaulus viviparus and Ostertagia ostertagi in Irish pasture-based dairy herds.

Infections with Dictyocaulus viviparus and Ostertagia ostertagi nematode parasites are of importance to bovine health and production in temperate areas across the world. Losses due to these parasites in dairy herds can be considerable due to decreased milk productivity and fertility. However, information on current epidemiological patterns in Irish dairy herds is limited. Bulk milk samples were collected from a total of 319 dairy farms across the Republic of Ireland. The D. viviparus samples were tested with an ELISA based on recombinant major sperm protein, while the O. ostertagi samples were tested with an ELISA based on crude saline extract, whole worm O. ostertagi antigen. Management data were collected from the farms using a questionnaire. Logistic regression was used to find significant associations between the presence of antibodies against D. viviparus and O. ostertagi and management factors. The overall prevalence of D. viviparus infection was 62.8%, while over 98% of herds had antibodies to O. ostertagi at the specified cut-off. Both D. viviparus and O. ostertagi antibodies were highest in November, which could be explained by the accumulated uptake of larvae through the grazing season. In herds of farmers that dosed their in-calf heifers with anthelmintics were significantly more likely to be positive for antibodies against D. viviparus infection. This study highlights that both D. viviparus and O. ostertagi infections are widespread in dairy herds in Ireland throughout the grazing season.

Comparison of two serum and bulk-tank milk ELISAs for diagnosing natural (sub)clinical Dictyocaulus viviparus infection in dairy cows.

Lungworm antibody ELISAs developed in Germany (DE) and The Netherlands (NL) were compared using four sets of serum (S) and bulk-tank milk (BTM) samples from adult dairy cows. The samples originated from 37 farms with or without a suspected clinical lungworm infection during August-October 2010 (Dataset 1), from cows excreting lungworm larvae or not during August-October 2010 (n=59) or May-June 2011 (n=164) (Dataset 2), from 305 farms in a national survey during October 2010 (Dataset 3), and 14 zero-grazing farms during February-April 2011 (Dataset 4). During August-October 2010, covering the second half of the grazing season, the NL-S and NL-BTM ELISA outperformed the DE-S and DE-BTM ELISAs in terms of sensitivity. For at least the NL-S and DE-S ELISA the opposite was found during May-June 2011, covering the end of the winter housing period and the early grazing season. Of the 305 farms in the survey 62.6% were found positive with the NL-BTM ELISA, whereas 2.6% was found positive with the DE-BTM ELISA. ODR values for the zero-grazing farms indicated that a cut-off value of 30% for the DE-BTM ELISA might be more appropriate than the currently used 41%. Results suggest that the NL ELISAs also respond to lungworm antigens other than Major Sperm Protein as used in the DE ELISAs. It is concluded that the generally higher sensitivity of the NL-BTM ELISA makes it better suited for large-scale prevalence studies and herd health monitoring programmes than the DE-BTM ELISA, positivity of which is more associated with the presence of clinical lungworm infection. Reducing the cut-off value of the DE-BTM ELISA from the original 49.3% to the current 41% or the possibly more appropriate 30% increased its sensitivity for detecting lungworm infections, but did not lead to similar sensitivity estimates as found for the NL-BTM ELISA.

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.