Riona Sayers

Benzimidazole carbamate residues in milk: Detection by Surface Plasmon Resonance-biosensor, using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method for extraction

A surface plasmon resonance (SPR) biosensor screening assay was developed and validated to detect 11 benzimidazole carbamate (BZT) veterinary drug residues in milk. The polyclonal antibody used was raised in sheep against a methyl 5(6)-[(carboxypentyl)-thio]-2-benzimidazole carbamate protein conjugate. A sample preparation procedure was developed using a modified QuEChERS method. BZT residues were extracted from milk using liquid extraction/partition with a dispersive solid phase extraction clean-up step. The assay was validated in accordance with the performance criteria described in 2002/657/EC. The limit of detection of the assay was calculated from the analysis of 20 known negative milk samples to be 2.7mugkg(-1). The detection capability (CCbeta) of the assay was determined to be 5mugkg(-1) for 11 benzimidazole residues and the mean recovery of analytes was in the range 81-116%. A comparison was made between the SPR-biosensor and UPLC-MS/MS analyses of milk samples (n=26) taken from cows treated different benzimidazole products, demonstrating the SPR-biosensor assay to be fit for purpose.

Factors associated with the concentration of immunoglobulin G in the colostrum of dairy cows

Transfer of sufficient immunoglobulin G (IgG) to the neonatal calf via colostrum is vital to provide the calf with immunological protection and resistance against disease. The objective of the present study was to determine the factors associated with both colostral IgG concentration and colostral weight in Irish dairy cows. Fresh colostrum samples were collected from 704 dairy cows of varying breed and parity from four Irish research farms between January and December 2011; colostral weight was recorded and the IgG concentration was determined using an ELISA method. The mean IgG concentration in the colostrum was 112 g/l (s.d. = 51 g/l) and ranged from 13 to 256 g/l. In total, 96% of the samples in this study contained >50 g/l IgG, which is considered to be indicative of high-quality colostrum. Mean colostral weight was 6.7 kg (s.d. = 3.6 kg) with a range of 0.1 to 24 kg. Factors associated with both colostral IgG concentration and colostral weight were determined using a fixed effects multiple regression model. Parity, time interval from calving to next milking, month of calving, colostral weight and herd were all independently associated with IgG concentration. IgG concentration decreased (P < 0.01) by 1.7 (s.e. = 0.6) g/l per kg increase in the colostral weight. Older parity cows, cows that had a shorter time interval from calving to milking, and cows that calved earlier in spring or in the autumn produced colostrum with higher IgG concentration. Parity (P < 0.001), time interval from calving to milking (P < 0.01), weight of the calf at birth (P < 0.05), colostral IgG concentration (P < 0.01) and herd were all independently associated with colostral weight at the first milking. Younger parity cows, cows milked earlier post-calving, and cows with lighter calves produced less colostrum. In general, colostrum quality of cows in this study was higher than in many previous studies; possible reasons include use of a relatively low-yielding cow type that produces low weight of colostrum, short calving to colostrum collection interval and grass-based nutritional management. The results of this study indicate that colostral IgG concentration can be maximised by reducing the time interval between calving and collection of colostrum.

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.

Investigation of the Persistence of Nitroxynil Residues in Milk from Lactating Dairy Cows by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

Nitroxynil is an anthelmintic used in the treatment of liver fluke. In this study, six dairy cows were treated during lactation with Trodax, a 34% solution containing nitroxynil as its N-ethylglucamine salt, indicated for the treatment of fascioliasis in cattle and sheep. Samples were collected twice daily for 16 days and later at weekly intervals up to 58 days post-treatment. Nitroxynil residues were extracted from milk samples using acetonitrile; magnesium sulfate and sodium chloride were added to induce liquid-liquid partitioning and purified by dispersive solid phase extraction for clean-up. Nitroxynil was determined by ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) in negative ionization mode. The limit of detection (CCα) of the method is 0.24 μg/kg. Maximum concentration of nitroxynil in the samples was in the range of 688-1358 μg/kg, with levels persisting for 58 days in four of the six lactating cows. Incurred nitroxynil samples were treated with sulfatase and β-glucuronidase from Helix pomatia ; the results indicated the presence of glucuronide conjugates in samples at early withdrawal times. At later withdrawal times the concentration of free nitroxynil was lower than the concentration in the control samples, indicating potential degradation during enzymatic treatment.

The single intradermal cervical comparative test interferes with Johne's disease ELISA diagnostics

Enzyme-linked immunosorbent assays (ELISA) of milk and serum samples are a routinely used method of screening herds for Mycobacterium avium subspecies paratuberculosis (MAP). Infection with MAP causes granulomatous enteritis of ruminants known as Johne’s disease (JD). The sensitivity (Se) and specificity (Sp) of MAP ELISAs leads to difficulties in the identification of both infected and infectious animals. Interference with MAP ELISA Se and Sp has been reported in MAP seronegative cows following administration of purified protein derivative (PPD) as part of intradermal testing for bovine tuberculosis (bTB). The aim of this study is to examine the impact of the single intradermal cervical comparative test (SICCT) for bTB, on both serum and milk MAP ELISA tests, in a herd containing both seropositive and seronegative cows pre-SICCT. A secondary objective is to provide appropriate timing of JD ELISA tests in relation to the SICCT. A herd of 139 cows were serum and milk sampled pre- and post-SICCT administration. Prior to SICCT, 6% of the herd tested seropositive for MAP using milk ELISA, with 8% positive on serum. ID Screen Paratuberculosis Indirect Screening Test (ID Vet) was used to screen the herd. Within 14 days of PPD administration, a significant increase in the prevalence of seropositive cows was recorded. Identical prevalence’s were recorded with both test matrices (39%). ELISA values remained significantly higher until day 43 post-SICCT in milk (P = 0.850), and day 71 in serum (P = 0.602). If the “new” positives detected post-bTB testing are deemed false positives due to generation of cross-reacting antibodies by administration of PPD, milk would appear a more suitable sample for JD ELISA testing within 2 months of SICCT. In summary, sampling for JD utilizing milk ELISA should be avoided in the 43-day period following PPD administration, with serum ELISA sampling avoided for an additional 28 days.

A survey of management practices on Irish dairy farms with emphasis on risk factors for Johne's disease transmission

BackgroundJohne’s disease (JD) is a chronic granulomatous enteritis affecting ruminants. A number of farm management practices are associated with increased risk of JD transmission. The aim of the current study was to document JD-related management practices currently employed on Irish dairy farms.Survey questions focused on calving area (CA), calf and manure management. Independent variables (region, calving-season, enterprise type, herd size and biosecurity status) were used to examine influences on JD associated dependent variables (survey questions). Additionally general biosecurity practices were also examined.ResultsResults showed management practices implemented by Irish dairy farmers pose a high risk of JD transmission. Of the farmers surveyed, 97% used the CA for more than one calving, 73.5% and 87.8% pooled colostrum and milk respectively, 33.7% never cleaned the CA between calving’s, and 56.6% used the CA for isolating sick cows. Survey results also highlighted that larger herds were more likely to engage in high risk practices for JD transmission, such as pooling colostrum (OR 4.8) and overcrowding the CA (OR 7.8). Larger herds were also less likely than smaller herds to clean the CA (OR 0.28), a practice also considered of risk in the transmission of JD.ConclusionMany management practices associated with risk of JD transmission were commonly applied on Irish dairy farms. Larger herds were more likely to engage in high risk practices for JD transmission. Control programmes should incorporate educational tools outlining the pathogenesis and transmission of JD to highlight the risks associated with implementing certain management practices with regard to JD transmission.

Questionnaire identifying management practices surrounding calving on spring-calving dairy farms and their associations with herd size and herd expansion.

Healthy calves are fundamental to any profitable dairy enterprise. Research to-date, has focused on year-round calving systems which experience many different challenges compared to spring-calving systems. The objective of the present study was to determine the on-farm dry cow, calving, and colostrum management practices of spring-calving dairy production systems, and quantify their associations with herd size and herd expansion status (i.e. expanding or not expanding). Information on these management practices was available from a survey of 262 Irish spring-calving dairy farmers, representative of the Irish national population. Herd expansion in the 2 years before, and the year that the survey was conducted was not associated with any of the management practices investigated. Fifty-three percent of respondents had an average calving season length of 10 to14 weeks with 35% of herds having a longer calving season. Previous research in cattle has documented that both colostrum source and feeding management are associated with the transmission of infectious disease from cow to calf. In the present study 60% of respondents fed calves colostrum from their own dam; however, 66% of those respondents allowed the calf to suckle the dam, 23% of survey respondents fed calves pooled colostrum. Larger herds were more likely (P<0.01) to use pooled colostrum supplies, while smaller herds were more likely (P<0.05) to allow the calf to suckle the dam. The majority (86%) of respondents had stored supplies of colostrum; average-sized herds had the greatest likelihood of storing colostrum (P<0.05), compared to other herd sizes; larger sized herds had a lesser likelihood (P<0.05) of storing colostrum in a freezer, compared to other herd sizes. Although freezing colostrum was the most common method used to store colostrum (54% of respondents), 17% of respondents stored colostrum at room temperature, 29% of which stored it at room temperature for greater than 4 days. The results from the present study indicate that a particular focus needs to be placed on calving and colostrum management because this study has highlighted a number of areas which are below international standards, and may have repercussions for calf health. Furthermore, management practices on larger farms could be improved and, as these represent the future of dairy farming, a focus needs to be placed on them. Expanding herds are not a particular concern as herd expansion, independent of herd size, does not seem to be associated with calving and colostrum management practices on Irish spring-calving dairy herds.

Effect of exposure to Neospora caninum, Salmonella, and Leptospira interrogans serovar Hardjo on the economic performance of Irish dairy herds

The objective of the current study was to quantify the effects of exposure to Salmonella, Neospora caninum, and Leptospira interrogans serovar Hardjo (L. hardjo) on dairy farm profitability and to simulate the effect of vaccination for Salmonella and L. hardjo on dairy farm profitability. The production effects associated with exposure to each of these pathogens in study herds were defined under 3 categories: (1) milk production effects, (2) reproduction effects (including culling), and (3) mortality effects. The production effects associated with exposure to Salmonella, N. caninum, and L. hardjo were incorporated into the Moorepark Dairy Systems Model. In the analysis, herds negative for exposure to Salmonella, N. caninum, and L. hardjo were assumed baseline herds, with all results presented relative to this base. In simulations examining the effect of vaccination for Salmonella and L. hardjo on farm profitability, vaccinated herds (vaccination costs included) were considered as baseline herds and results were presented relative to this base. Total annual profits in unvaccinated herds were reduced by €77.31, €94.71, and €112.11 per cow at milk prices of €0.24, €0.29, and €0.34/L, respectively, as a result of exposure to Salmonella. In the current study, herds positive for exposure to Salmonella recorded a 316-kg reduction in milk yield, whereas no association was detected between exposure to N. caninum or L. hardjo and milk production. Exposure to both N. caninum and L. hardjo was associated with compromised reproductive performance. Herds positive for exposure to N. caninum and Salmonella had greater rates of adult cow mortality and calf mortality, respectively. Vaccination for both Salmonella and L. hardjo was associated with improved performance in study herds. Exposure to N. caninum resulted in a reduction in annual farm profits of €11.55, €12, and €12.44 per cow at each milk price, whereas exposure to L. hardjo resulted in a reduction in annual farm profits of €13.83, €13.78, and €13.72 per cow at each milk price. Herds that tested positive for exposure to Salmonella and L. hardjo were compared with herds vaccinated for the respective pathogens. Herds vaccinated for Salmonella generated €67.09, €84.48, and €101.89 per cow more profit at each milk price compared with herds positive for exposure. Similarly, herds vaccinated for L. hardjo generated €9.74, €9.69, and €9.63 per cow more profit compared with unvaccinated exposed herds. However, herds that tested negative for exposure to Salmonella and L. hardjo generated additional profits of €10.22 and €4.09 per cow, respectively, compared with vaccinated baseline herds.

Genetic parameters for both a liver damage phenotype caused by and antibody response to phenotype in dairy and beef cattle

is a helminth parasite of economic importance to the global cattle industry, with documented high international herd prevalence. The objective of the present study was to generate the first published genetic parameter estimates for liver damage caused by as well as antibody response to in cattle. Abattoir data on the presence of live , or -damaged livers, were available between the years 2012 and 2015, inclusive. A second data set was available on cows from 68 selected dairy herds with a blood ELISA test for antibody response to in autumn 2015. Animals were identified as exposed by using herd mate phenotype, and only exposed animals were retained for analysis. The abattoir data set consisted of 20,481 dairy cows and 75,041 young dairy and beef animals, whereas the study herd data set consisted of 6,912 dairy cows. (Co)variance components for phenotypes in both data sets were estimated using animal linear mixed models. Fixed effects included in the model for both data sets were contemporary group, heterosis coefficient, recombination loss coefficient, parity, age relative to parity/age group, and stage of lactation. An additional fixed effect of abattoir by date of slaughter was included in the model for the analysis of the abattoir data. Direct additive genetic effects and a residual effect were included as random effects for all analyses. After data edits, the prevalence of liver damage caused by in cows and young cattle was 47% and 20%, respectively. The prevalence of a positive antibody response to in cows from the study herd data was 36% after data edits. The heritability of as a binary trait for dairy cows in abattoir data and study herd data was 0.03 ± 0.01 and 0.09 ± 0.02, respectively; heritability in young cattle was 0.01 ± 0.005. The additive genetic SD of as a binary trait was 0.069 and 0.050 for cows and young cattle from the abattoir data, respectively, and 0.112 from the study herd cows. The genetic correlation between liver damage caused by in young cattle and cows from the abattoir data was 0.94 ± 0.312 and the genetic correlation between liver damage caused by in cows and positive antibody response to in cows in the study herd data was 0.37 ± 0.283.

Associations between exposure to bovine herpesvirus 1 (BoHV-1) and milk production, reproductive performance, and mortality in Irish dairy herds

As cost-benefit analyses are required to prioritize and promote disease control and eradication programs within a jurisdiction, national data relating to disease-related production losses are particularly useful. The objectives of the current study were to use Irish bovine herpesvirus 1 (BoHV-1) prevalence data in dairy herds, obtained by bulk milk sampling on 4 occasions over the 2009 lactation, to document associations between milk production, fertility performance, mortality, and BoHV-1 herd status. Bulk milk (n = 305) antibody ELISA was used to classify farms as positive or negative in terms of endemic BoHV-1. Cow-level (milk parameters only) and herd-level performance data were sourced from the Irish Cattle Breeding Federation. Ordinary linear and negative binomial regressions were used to investigate associations between milk, fertility, and mortality performance and herd-level BoHV-1 results (both categorical and continuous variables). Only slight effects on the rates of carryover cows, nonpregnant cows, and total deaths were highlighted with increasing ELISA sample/positive (%) values (incidence rate ratio = 1.001). Multiparous cows in herds BoHV-1 bulk milk antibody positive recorded a reduction in milk yield per cow per year of 250.9 L in the multivariable linear model. Milk fat and protein yields were also affected by herd BoHV-1 status, again highlighting sub-optimal milk production in BoHV-1 bulk milk-positive herds. The current study has highlighted an economical method of investigating losses due to endemic infection using repeated bulk milk sampling over a single lactation. These data can contribute to analyzing the cost-benefit of applying BoHV-1 control strategies both on farm and at a national level.