This Research Paper addresses the hypothesis that the dietary inclusion of an Aspergillus niger fermentation product will alter the degradation kinetics and rumen fermentation patterns of feeds in dairy cattle. Fungal fermentation products often contain a suite of bioactive compounds and secondary metabolites, which can influence the microbial environment in the rumen and act as digestibility enhancers. As the cattle sector is under increasing pressure to enhance its sustainability, the investigation of dietary interventions that could improve the efficiency of production is warranted. In a previous experiment, Synergen®, a product of the solid-state fermentation of Aspergillus niger (ANP) containing residual enzyme activities, significantly increased the in vitro digestibility of a grass silage-based dairy total mixed ration (TMR), suggesting that in vivo studies would be valuable. Hence the present study aimed to quantify the effects of this ANP on rumen fermentation measures in cattle. Using a 4 × 4 Latin square design, the effect of four doses of ANP (0, 5, 10, 15 g/day) in four cannulated Jersey heifers was measured on the in sacco degradation of dry matter (DM), organic matter, crude protein and neutral detergent fibre in steam-flaked barley, grass silage and a grass silage-based TMR formulated for dairy cattle. Treatments had no significant effect on the rate, or extent, of degradation of any component in any feed investigated. Rumen volatile fatty acid concentrations and proportions, and rumen pH, were quantified at seven timepoints during each 48-h sampling period and were unaffected by treatment, as was the apparent total tract digestibility of DM. Under the conditions of this trial, ANP did not influence rumen fermentation kinetics; indicating that supplementing mature, non-lactating Jersey cattle with this fungal fermentation product is not an advantageous strategy to enhance feed digestibility.

The skin is the primary feeding site of ticks that infest livestock animals such as cattle. The highly specialised functions of skin at the molecular level may be a factor contributing to variation in susceptibility to tick infestation; but these remain to be well defined. The aim of this study was to investigate the bovine skin transcriptomic profiles of tick-naïve and tick-infested cattle and to uncover the gene expression networks that influence contrasting phenotypes of host resistance to ticks. RNA-Seq data was obtained from skin of Brangus cattle with high (n = 5) and low (n = 6) host resistance at 0 and 12 weeks following artificial tick challenge with Rhipicephalus australis larvae. No differentially expressed genes were detected pre-infestation between high and low resistance groups, but at 12-weeks there were 229 differentially expressed genes (DEGs; FDR < 0.05), of which 212 were the target of at least 1866 transcription factors (TFs) expressed in skin. Regulatory impact factor (RIF) analysis identified 158 significant TFs ( P < 0.05) of which GRHL3 , and DTX1 were also DEGs in the experiment. Gene term enrichment showed the significant TFs and DEGs were enriched in processes related to immune response and biological pathways related to host response to infectious diseases. Interferon Type 1-stimulated genes, including MX2 , ISG15 , MX1 , OAS2 were upregulated in low host resistance steers after repeated tick challenge, suggesting dysregulated wound healing and chronic inflammatory skin processes contributing to host susceptibility to ticks. The present study provides an assessment of the bovine skin transcriptome before and after repeated tick challenge and shows that the up-regulation of pro-inflammatory genes is a prominent feature in the skin of tick-susceptible animals. In addition, the identification of transcription factors with high regulatory impact provides insights into the potentially meaningful gene–gene interactions involved in the variation of phenotypes of bovine host resistance to ticks.

Bovine erythropoietic protoporphyria (BCEPP) is a rare genetic disorder predominantly affecting Limousin and, sporadically, Blonde Aquitaine cattle. It arises from diminished or absent ferrochelatase activity, causing the toxic buildup of protoporphyrin in tissues. This accumulation leads to photosensitivity and seizures upon exposure to sunlight. Here, we report a case of a female Limousin calf exhibiting BCEPP signs, including photosensitivity, skin lesions, ataxia, and seizure. Clinical examination and ancillary tests ruled out photosensitivity related to direct ingestion or hepatogenous toxins. Photosensitivity due to aberrant porphyrin accumulation, specifically BCEPP and Congenital Erythropoietic Porphyria (CEP), emerged as the primary differential diagnoses. Notably, the absence of characteristic signs such as "pink teeth" and anaemia, which are indicative of CEP, distinguished BCEPP. A homozygous autosomal recessive mutation in the ferrochelatase gene (FECH) confirmed the BCEPP diagnosis by genetic testing. The animal's welfare led to euthanasia and subsequent farm control strategies aimed at preventing further genetic transmission. To eliminate the chance of transmitting the genetic mutation, the decision was made to switch to a pure Aberdeen Angus bull. In conclusion, this case emphasises the significance of a comprehensive diagnostic approach. The farm history, clinical examination, and ancillary testing helped to narrow down other differential diagnoses further. The distinction between BCEPP and CEP rested on observable clinical indicators and breed specificity. The targeted genetic testing confirmed BCEPP paving the way for the correct herd health approach.

A wide spectrum of disease severity associated with cryptosporidiosis has been described, ranging from asymptomatic to fatal in both human and animal hosts. The reasons for the variations in severity are likely to be multifactorial, involving environmental, host and parasite factors. This paper describes two experimental infection trials in lambs, a symptomatic host for the parasite, to investigate variation in the clinical manifestations following infection with two distinct isolates of Cryptosporidium parvum. In the first experiment, groups of naïve lambs were challenged with one of two isolates (CP1 or CP2) at ​< ​1 week of age, to test the effect of the isolates on disease outcome. In a second experiment one group of lambs challenged at < 1 week of age (CP1) was then re-challenged with the same isolate at 6 weeks of age (CP1), while a second group was challenged for the first time at 6 weeks of age (CP1). This experiment examined age-related disease symptoms, oocyst shedding and the effect of prior exposure to the parasite on a subsequent homologous challenge. The two isolates were associated with significant differences in the demeanour of the animals and in the numbers of oocysts shed in the faeces. There were also differences in the duration and severity of diarrhoea, though these were not significant. The age of the lamb, at the time of a primary challenge (<1 week or 6 weeks), also resulted in differences in clinical outcomes, with younger lambs showing more severe clinical disease than the older lambs (feeding profiles and presentation of diarrhoea), while older lambs showed virtually no signs of infection but still produced large numbers of oocysts.

AbstractThis observational study aimed to describe the diurnal pattern of reticuloruminal contraction rate (RRCR) and the proportion of time spent ruminating by cattle, using two commercial devices equipped with triaxial accelerometers: an indwelling bolus (placed in the reticulum) and a neck collar. The three objectives of this study were firstly to determine whether the indwelling bolus provided observations consistent with RRCR as determined by clinical examination using auscultation and ultrasound, secondly to compare estimates of time spent ruminating using the indwelling bolus and a collar-based accelerometer, and finally to describe the diurnal pattern of RRCR using the indwelling bolus data. Six rumen-fistulated, non-lactating Jersey cows were fitted with an indwelling bolus (SmaXtec Animal Care GmbH, Graz, Austria) and a neck collar (Silent Herdsman, Afimilk Ltd. Kibbutz Afikim, Israel), and data were collected over two weeks. Cattle were housed together in a single straw-bedded pen and fed ad libitum hay. To assess the agreement between the indwelling bolus and traditional methods of assessing reticuloruminal contractility in the first week, the RRCR was determined over 10 min, twice a day, by ultrasound and auscultation. Mean inter-contraction intervals (ICI) derived from bolus and ultrasound, and from auscultation were 40.4 ± 4.7, 40.1 ± 4.0 and 38.4 ± 3.3 s. Bland–Altmann plots showed similar performance of the methods with small biases. The Pearson correlation coefficient for the time spent ruminating derived from neck collars and indwelling boluses was 0.72 (highly significant, P < 0.001). The indwelling boluses generated a consistent diurnal pattern for all the cows. In conclusion, a robust relationship was observed between clinical observation and the indwelling boluses for estimation of ICI and, similarly, between the indwelling bolus and neck collar for estimating rumination time. The indwelling boluses showed a clear diurnal pattern for RRCR and time spent ruminating, indicating that they should be useful for assessing reticuloruminal motility.

IntroductionBreeding for tick resistance is a sustainable alternative to control cattle ticks due to widespread resistance to acaricidal drugs and the lack of a protective vaccine. The most accurate method used to characterise the phenotype for tick resistance in field studies is the standard tick count, but this is labour-intensive and can be hazardous to the operator. Efficient genetic selection requires reliable phenotyping or biomarker(s) for accurately identifying tick-resistant cattle. Although breed-specific genes associated with tick resistance have been identified, the mechanisms behind tick resistance have not yet been fully characterised.MethodsThis study applied quantitative proteomics to examine the differential abundance of serum and skin proteins using samples from naïve tick-resistant and -susceptible Brangus cattle at two-time points following tick exposure. The proteins were digested into peptides, followed by identification and quantification using sequential window acquisition of all theoretical fragment ion mass spectrometry.ResultsResistant naïve cattle had a suite of proteins associated with immune response, blood coagulation and wound healing that were significantly (adjusted P < 10- 5) more abundant compared with susceptible naïve cattle. These proteins included complement factors (C3, C4, C4a), alpha-1-acid glycoprotein (AGP), beta-2-glycoprotein-1, keratins (KRT1 & KRT3) and fibrinogens (alpha & beta). The mass spectrometry findings were validated by identifying differences in the relative abundance of selected serum proteins with ELISA. The proteins showing a significantly different abundance in resistant cattle following early and prolonged tick exposures (compared to resistant naïve) were associated with immune response, blood coagulation, homeostasis, and wound healing. In contrast, susceptible cattle developed some of these responses only after prolonged tick exposure.DiscussionResistant cattle were able to transmigrate immune-response related proteins towards the tick bite sites, which may prevent tick feeding. Significantly differentially abundant proteins identified in this research in resistant naïve cattle may provide a rapid and efficient protective response to tick infestation. Physical barrier (skin integrity and wound healing) mechanisms and systemic immune responses were key contributors to resistance. Immune response-related proteins such as C4, C4a, AGP and CGN1 (naïve samples), CD14, GC and AGP (post-infestation) should be further investigated as potential biomarkers for tick resistance.

There has been little information about the proteome of bovine faeces or about the contribution to the faecal proteome of proteins from the host, the feed or the intestinal microbiome. Here, the bovine faecal proteome and the origin of its component proteins was assessed, while also determining the effect of treating barley, the major carbohydrate in the feed, with either ammonia (ATB) or sodium propionate (PTB) preservative. Healthy continental crossbreed steers were allocated to two groups and fed on either of the barley-based diets. Five faecal samples from each group were collected on Day 81 of the trial and analysed by quantitative proteomics using nLC-ESI-MS/MS after tandem mass tag labelling. In total, 281 bovine proteins, 199 barley proteins, 176 bacterial proteins and 190 archaeal proteins were identified in the faeces. Mucosal pentraxin, albumin and digestive enzymes were among bovine proteins identified. Serpin Z4 a protease inhibitor was the most abundant barley protein identified which is also found in barley-based beer, while numerous microbial proteins were identified, many originating bacteria from Clostridium, while Methanobrevibacter was the dominant archaeal genus. Thirty-nine proteins were differentially abundant between groups, the majority being more abundant in the PTB group compared to the ATB group. SIGNIFICANCE: Proteomic examination of faeces is becoming a valuable means to assess the health of the gastro-intestinal tract in several species, but knowledge on the proteins present in bovine faeces is limited. This investigation aimed to characterise the proteome of bovine faecal extracts in order to evaluate the potential for investigations of the proteome as a means to assess the health, disease and welfare of cattle in the future. The investigation was able to identify proteins in bovine faeces that had been (i) produced by the individual cattle, (ii) present in the barley-based feed eaten by the cattle or (iii) produced by bacteria and other microbes in the rumen or intestines. Bovine proteins identified included mucosal pentraxin, serum albumin and a variety of digestive enzymes. Barley proteins found in the faeces included serpin Z4, a protease inhibitor that is also found in beer having survived the brewing process. Bacterial and archaeal proteins in the faecal extracts were related to several pathways related to the metabolism of carbohydrates. The recognition of the range of proteins that can be identified in bovine faeces raises the possibility that non-invasive sample collection of this material could provide a novel diagnostic approach to cattle health and welfare.

Background Hypohidrotic ectodermal dysplasia (HED) is a congenital syndrome of mammals affecting organs and tissues of ectodermal origin characterized by absence or hypoplasia of hair, teeth, and eccrine glands. The disorder has been reported in several species, including humans, mice, dogs and cattle, associated with variants in genes affecting the ectodysplasin pathway, including the X-linked ectodysplasin A (EDA) gene. Until now, nine pathogenic variants have been found in the bovine EDA gene. Here we report a novel variant in EDA in a crossbreed male Belgian Blue calf with HED, and provide an overview of the phenotypic and allelic heterogeneity of EDA-related forms of HED in cattle. Case presentation A 45-day-old male crossbreed British Blue calf was referred with congenital hypotrichosis, oligodontia and omphalitis. On histopathological examination of the nasal planum, nasolabial glands and ducts were not observed. The density of hair follicles was low, and they were small, with a predominance of telogen-phase hairs, and some serocellular crusts. The phenotype of the calf resembled that of HED. Whole-genome sequencing (WGS) was performed and revealed a 21,899 base-pair deletion encompassing the coding exon 2 of EDA, predicted to result in an altered transcript and aberrant protein. Conclusions The clinicopathological and genetic findings were consistent with a case of X-linked HED. A very similar EDA deletion has been previously reported in a family of Holstein cattle with HED. The newly identified hemizygous EDA loss-of-function variant is certainly pathogenic and therefore is the genetic cause for the observed phenotype. This case report provides an additional example of the potential of WGS-based precise diagnostics in livestock species such as cattle to increase the diagnostic yield in rare diseases.

High-moisture cereal grains are often treated with ammonia to prevent spoilage and for perceived animal performance advantages, but there are few reports on the effects of ammonia treatment of cereal on cattle performance. This study was carried out on 101 Charolais cattle to quantify the effects of feeding ammonia-treated maize on the performance of intensively fattened beef cattle. Animals were assigned to 16 pens in two sheds and all animals in each pen were fed on the same diet for 176 d. The cereal in the diets was either ammonia-treated maize (ATM) or untreated maize diet (UTM), formulated to be approximately isoenergetic and isonitrogenous. Cattle fed ATM had a lower feed (c)onversion ratio (FCR) (6.14 +/- 0.39 vs 6.49 +/- 0.41 kg DM/kg BW; p = 0.011) and tended to have a higher average daily liveweight gain (ADG) (1.60 +/- 0.10 vs 1.54 +/- 0.10 kg/d; p = 0.060). The pH of the ruminal fluid at slaughter was higher (p < 0.001) in animals that were fed the ATM diet (6.35 +/- 0.69) compared to those fed the UTM diet (5.46 +/- 0.33). No differences were found in the concentrations of NH3 or total VFA in ruminal fluid between the two groups. The molar proportion of butyric acid from all VFAs was lower in animals fed ATM than UTM (14.69 +/- 3.55% and 18.04 +/- 2.97% respectively; p = 0.012), and the molar proportion of propionic acid was higher in the ATM than the UTM groups (22.14 +/- 1.54% and 19.54 +/- 1.72% respectively; p < 0.001). The ratio of acetic acid to propionic acid was significantly lower in animals fed ATM compared to those fed UTM (ATM vs UTM: 2.64 +/- 0.26 vs 2.97 +/- 0.29; p = 0.0038). Faecal starch was lower in cattle fed ATM than untreated maize (UTM) (14.73 +/- 5.65 vs 18.14 +/- 4.27% DM; p < 0.001). The pH of faeces in the ATM group was higher than that in the UTM group (4.75 +/- 0.19 vs 4.55 +/- 0.19; p < 0.001). In conclusion, ammonia treatment of maize resulted in increased ADG and decreased FCR, with reduced faecal starch concentration, suggesting possible gains in efficiency of starch utilisation.

Bovine faecal composition is complex and a knowledge gap exists in the understanding of the bovine faecal proteome. In the present study, in-gel sample preparation (IGSP) of faecal samples prior to proteomics showed an increase in the number of proteins identified in faecal samples compared to those processed by filter-aided sample preparation (FASP). The optimised sample preparation method removed high molecular weight glycoproteins as part of the clean-up process of the faecal samples, and in combination with in-gel digestion before liquid chromatography with tandem mass spectrometry (LC-MS/MS). The use of IGSP led to enhanced protein identification with increases in the number of peptides identified and in the percent coverage of proteins in the bovine faecal samples. SIGNIFICANCE: Characterization of faecal proteins has the potential to increase our understanding of host responses to changes such as diet, disease and drug-treatment. In-gel sample preparation prior to proteomics can be used to remove high molecular weight glycoproteins and reduce protein/peptide loss in FASP. This method of sample preparation will have application not only in the investigation of bovine faecal extracts but also in studies where large molecules such as glycoproteins or oligosaccharides could have detrimental influences on sample preparation involving ultrafiltration.

Background Disease emergence and production loss caused by cattle tick infestations have focused attention on genetic selection strategies to breed beef cattle with increased tick resistance. However, the mechanisms behind host responses to tick infestation have not been fully characterised. Hence, this study examined gene expression profiles of peripheral blood leukocytes from tick-naive Brangus steers ( Bos taurus x Bos indicus ) at 0, 3, and 12 weeks following artificial tick challenge experiments with Rhipicephalus australis larvae. The aim of the study was to investigate the effect of tick infestation on host leukocyte response to explore genes associated with the expression of high and low host resistance to ticks. Results Animals with high (HR, n = 5) and low (LR, n = 5) host resistance were identified after repeated tick challenge. A total of 3644 unique differentially expressed genes (FDR < 0.05) were identified in the comparison of tick-exposed (both HR and LR) and tick-naive steers for the 3-week and 12-week infestation period. Enrichment analyses showed genes were involved in leukocyte chemotaxis, coagulation, and inflammatory response. The IL-17 signalling, and cytokine-cytokine interactions pathways appeared to be relevant in protection and immunopathology to tick challenge. Comparison of HR and LR phenotypes at timepoints of weeks 0, 3, and 12 showed there were 69, 8, and 4 differentially expressed genes, respectively. Most of these genes were related to immune, tissue remodelling, and angiogenesis functions, suggesting this is relevant in the development of resistance or susceptibility to tick challenge. Conclusions This study showed the effect of tick infestation on Brangus cattle with variable phenotypes of host resistance to R. australis ticks . Steers responded to infestation by expressing leukocyte genes related to chemotaxis, cytokine secretion, and inflammatory response. The altered expression of genes from the bovine MHC complex in highly resistant animals at pre- and post- infestation stages also supports the relevance of this genomic region for disease resilience. Overall, this study offers a resource of leukocyte gene expression data on matched tick-naive and tick-infested steers relevant for the improvement of tick resistance in composite cattle.

When dairy cows produce little or no colostrum, calves are likely to suffer from failure of passive transfer (FPT). Volume of colostrum produced by the dam may be affected by: nutrition; environmental conditions; time from calving to milking; parity; dry period length; calving difficulty; calf weight; calf sex; calf viability; cow BCS / body weight; milk production in previous lactation and dam health. While risk factors for poor IgG concentration in colostrum have been extensively studied, there is little published literature on low colostrum yields and associated risk factors. The aim of this observational study was to identify risk factors for poor colostrum yield in dairy cows and to quantify the effect of variation in those variables that the farmer can directly control (length of dry period, and close-up transition period nutrition) using inverse probability modelling. 439 animals from a single Scottish dairy farm were used as a convenience sample. First milking colostrum yield was measured by farmers using a graduated bucket. For each animal, risk factor data and meteorological data from the nearest weather were obtained. Decreased colostrum yield was associated with increasing daily temperature range, while increasing UV index, lactation number and number of days dry increased colostrum yield. Marginal effect models revealed that for each additional day of dry period, colostrum yield increased. Higher UV indices the month before calving were positively associated with colostrum yield, particularly in primiparous cows, and higher temperatures in the months before calving were positively associated with colostrum yield in all cows.

Tick resistance is a desirable trait in beef cattle to minimise the incidence of cattle tick outbreaks and production loss in tropical and subtropical regions. Identification of genetic determinants of this trait will be beneficial for improved breeding programs. Using RNA-Sequencing, this study investigated gene expression data from skin of tick-infested Brangus cattle with high (n=5) and low (n=6) resistance. There were 229 significant differentially expressed genes (DEGs; FDR<0.05), of which 212 were target genes for 1,866 transcription factors expressed in skin. Regulatory impact factor analysis showed 158 significant TFs (P<0.05) of which GRHL3, and DTX1 were also DEGs in the experiment. Biological pathway enrichment showed the significant TFs and DEGs feature in pathways such as response to infection, transcriptional regulation, and Notch signalling. These results highlight the application of bioinformatic approaches in the identification of key transcriptional factor that are potential drivers of differential expression of host resistance trait in cattle. Thus, these candidate genes could be further explored in gene selection programs to breed for tick resistance in crossbred cattle.

High levels of supplementation with cereal increases production rates in cattle but can increase incidence of disease, ranging from mild indigestion to acute ruminal acidosis and death. Therefore, there is motivation to determine biological markers which can be used to identify whether animals have been, or are being fed, sufficient or excessive cereals. This study aimed to describe light microscopic findings from animals being fed diverse dietary cereal proportions and to test the performance of a novel rumen epithelial scoring system. Rumen wall tissue samples were obtained from the abattoir from 195 cattle from 11 Scottish farms and processed for histological examination. Light microscopic examination was used to characterise ruminal epithelial response to dietary challenge. Secondary objectives included describing the distribution of immune-related cells in bovine ruminal epithelium and assessing the use of a modified Elastin Martius Scarlet Blue stain (EMSB) for histological examination of the rumen epithelium. Cells staining positive for cluster of differentiation 3 were distributed mainly in the lower layers of the stratum basale and were found in higher densities in animals offered lower cereal proportion diets. Cells staining positive for major histocompatibility complex class 2 (MHCII) were most common in perivascular locations and in the junction between the lower stratum basale and the propria-submucosa. The density of MHCII positive staining cells was higher in animals on lower cereal diets. The level of supplementation with cereal was also associated with the thickness of the stratum corneum (SCT) and stratum granulosum (SGT), the integrity of the stratum corneum and sloughing of cornified cells. There were no advantages in using EMSB stain over haematoxylin and eosin (H&E) in this scoring system. We concluded that a scoring system that included only SCT, SGT and a measure of the loss of appearance of intercellular space allowed differentiation of groups of animals according to the level of cereal supplementation.

Although Nematodirus battus is a serious threat to the health and survival of young lambs, there are few options to control this parasite. Bayesian Monte Carlo Markov Chain modelling with a zero-inflated Poisson distribution was used to estimate the heritability of egg counts in both June and July for each of five consecutive cohorts of 200 Scottish Blackface lambs. In one of the 10 analyses, the results failed the diagnostic tests. In seven of the analyses, there was no convincing evidence that the variation in egg counts was heritable. In the 2 years of high infection, the heritability was approximately 0.4 in June but the estimates lacked precision and the 95% highest posterior density credible intervals ranged from just above zero to 0.7. Selective breeding for resistance to N. battus will be difficult because genetically resistant or susceptible lambs cannot be consistently identified by phenotypic markers.

We aimed to estimate 1) the marginal effect of liver fluke (Fasciola hepatica) infection on productivity of Scottish beef cattle, and 2) the associated greenhouse gas emissions intensity (GHG EI). Data comprised 240,065 abattoir records from NE Scotland from 2014 to 2017, including the presence or absence of lesions typical of liver fluke in the liver at the time of slaughter, from which we inferred liver fluke infection status. The retrospective analysis of abattoir records to estimate marginal effects of an exposure is complicated by the multi-dimensional, clustered nature of the datasets, which result in confounding of potential causal factors with the exposure. Causal inference methods are required to identify and correct for variation in background exposure. We constructed directed acyclic graphs (DAGs) of observed variables, including the potential confounders, breed, sex, breeder, finisher, season of birth and year of birth. We then applied inverse probability weighting (IPW) to adjust for variation among exposure risk and applied a doubly robust generalized linear model (DRGLM) to the weighted observations to estimate the marginal effect of fluke on the growth rate of animals and total days from birth until slaughter. We compared these estimates with the results of linear mixed effects (LME) models with the same variables, treating breeder and producer as random effects. To estimate GHG EI, we applied IPCC tier-2 type GHG calculations to the marginal effects estimated from IPW with DRGLM. The IPW with DRGLM model estimated that animals with active fluke lesions (adult fluke seen on postmortem inspection) gained 17 (95 % CI 12-22) g/d less saleable beef than animals with no lesions and no visible fluke. Animals with active fluke lesions were 11 (95 % CI 6.5-15) d older at slaughter weight than animals with no lesions. Animals with historic lesions in which there was scarring of the liver but in which no adult fluke were seen showed a wide variation in effect estimates, consistent with some misclassification. The effect estimates from LME models suggested slightly lower effects of fluke on growth rate and days to slaughter but with overlapping 95 % confidence intervals. Calculation of the associated GHG emissions suggest the EI of meat from a herd with no fluke is approximately 1.5 % lower than the same herd with fluke. Sustainably controlling liver fluke would have additional production benefits not included in this estimate and could therefore have a much greater impact on GHG EI in practice than demonstrated here.