Effect of source and journey on physiological variables in calves transported by road and ferry between Ireland and the Netherlands.

This study aimed to establish baseline variables for calves transported by road and ferry from Ireland to the Netherlands and to investigate the effect of journey [two comparable journeys in April (J1) and May (J2) 2022] and source [source farm or mart (SF/MA)] on these variables. A total of 66 calves from the SF/MA were transported from Ireland to commercial veal farms in the Netherlands. Blood samples were collected at the SF/MA, assembly center (Ireland), lairage (France), and on arrival on the veal farm (Netherlands). They were analyzed for indicator variables related to energy balance, hydration/electrolytes, physical/muscular stress, immunity, and inflammation [glucose, beta-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), potassium, sodium, magnesium, chloride, urea, haematocrit, total protein, creatine kinase, L-lactate, cortisol, white blood cell, neutrophil, lymphocyte and monocyte counts, serum amyloid-A, and haptoglobin]. Health variables eye and nose discharge, skin tent (a measure of dehydration), and navel inflammation were scored by a trained observer, and calves were weighed at every blood-sampling time point. All blood variables and body weight changed significantly (P < 0.05) during transport, most notably between the assembly center and lairage. Reference ranges were available for 18 variables; 11 of these variables exceeded the reference ranges at the lairage, whilst 10 variables exceeded the reference ranges on arrival at the veal farm. However, health variables did not change during transport. A journey-to-journey comparison indicated much variation; 18 out of 25 variables differed significantly on at least one time point. In total, J1 calves experienced a more severe change in BHB, potassium, strong-ion-difference, L-lactate, and eye and nose discharge than J2 calves. The source of calves also affected their physiology; 12 out of 25 variables studied differed significantly, all of which were confined to the first time point. Specifically, MA calves had elevated levels of NEFA, urea, haematocrit, L-lactate, cortisol, white blood cell, neutrophil, and monocyte counts and lower levels of corrected chloride and lymphocyte count. Overall, calves in this study showed a generalized physiological disturbance beyond reference limits during long-distance transport, but no animal died during transport or for 3 weeks post-arrival.

The effect of slow-release milk replacer feeding on health and behaviour parameters in dairy breed calves.

The aim of this research paper was to evaluate the effect of a slow-release milk replacer on health and behaviour of neonatal dairy calves. This was done with the potential benefits to welfare during transport in mind. A total of 15 calves were randomly divided into three groups of 5, namely, a control group fed twice in 24 h with 3 l of a conventional milk replacer, a slow-release group fed once in 24 h with 2 l of conventional milk replacer and 1 litre of a specialised micro-encapsulated feed and an enriched-replacer group fed once in 24 h with 3 l of milk replacer enriched with micellar casein. Blood samples were taken before feeding and 6, 12, 18 and 24 h after and analysed for acid-base parameters, electrolytes, glucose, haemoglobin, cortisol, insulin, cholecystokinin and adiponectin. Calf behaviour was recorded between 6 and 14 h after feeding. There was a significant increase in blood pH 6 h after feeding in all groups, but the glucose, HCO3 - and base excess increased significantly in the slow-release group only, whereas sodium increased significantly in the enriched group only. Glucose levels remained significantly higher in the slow-release group, relative to the control, at 6, 12, and 18 h after feeding. Insulin levels changed significantly over time in the enriched and control group but remained constant in the slow-release group. Insulin levels were significantly higher in the control group when compared to the slow-release group after feeding. Adiponectin changed significantly over time after feeding in the control group only, but no significant changes were observed between the feeding groups. Behavioural patterns were similar in control and slow release groups but less favourable (less lying time, more vocalisations) in the enriched group. In conclusion, once-daily feeding of slow-release milk replacer demonstrated favourable patterns of blood variables related to satiety and hunger as well as behavioural patterns that did not differ from conventional twice-daily feeding.

Evaluation of the optimal strong ion difference concentration of an oral electrolyte and buffering solution for the treatment of neonatal calf diarrhea.

An observational study was conducted to comparatively assess the efficacy of three different oral rehydration and buffering solutions, differentiated by their strong ion difference (SID) concentration, for treatment of neonatal calves with naturally acquired diarrhea. The SID concentrations tested were 100 mM, 170 mM and 230 mM for treatments SID100, SID170 and SID230, respectively. Clinical assessment and blood gas analysis were completed for 18 diarrheic calves once pre- and twice post- (6 and 24 hr after) oral administration with one of the three treatments. A repeated measure mixed model approach was used to analyze (a) the within-group efficacy of each treatment over time and (b) the between-group comparison at each timepoint. SID230 treatment resulted in a significant increase in blood pH, HCO3-, BE, SID and Na+ at 6 and 24 hr after treatment, and a significant decrease in AG and K+ by 24 hr after treatment. There were no significant changes in any of the blood gas parameters after treatment with SID100 and SID170. SID230 treatment also resulted in blood gas parameter changes that were significantly different to the other two groups. These results suggest that the optimum SID concentration for the treatment of calves with diarrhea is likely to be higher than current recommendations.

Determining the predictive capability of a Clinical Assessment Scoring Chart to differentiate severity of the clinical consequences of neonatal calf diarrhea relative to gold-standard blood gas analysis.

Neonatal calf diarrhea (NCD) is a major problem to calf health worldwide, in terms of both morbidity and mortality. A five-point ordinal scale clinical assessment scoring (CAS) chart was utilized to assess calves suffering from NCD-related clinical abnormalities (acidosis and dehydration) on commercial farms. The objective of this research was to determine the predictive capability of this CAS chart against gold standard blood gas parameters, designed to assist farmers in the accurate assessment of the clinical consequences of NCD. A total of 443 diarrheic and non-diarrheic calves were enrolled in the study. The CAS chart rated a calf's health from no clinical signs to varying degrees of clinical severity on a 0 (clinically normal) to 4 (grave) scale, based on clinical indicators including calf demeanour, ear position, mobility, suckle reflex, desire-to-feed, and enophthalmos. Blood gas analysis was conducted for individual calves, consisting of pH, base excess, Na+, K+, Ca2+, Cl-, glucose, total hemoglobin, bicarbonate, anion gap, and strong ion difference. Statistical evaluation was performed by comparison of the CAS score with blood gas profiles using ordinal logistic regression and a non-parametric estimation of the Receiver Operating Characteristics (ROC). The ROC analysis indicated that the CAS chart had acceptable specificity (>95%) with low sensitivity (<60%) in differentiating clinically normal from acidotic/dehydrated cases. Assessment of individual severity classes indicated that the chart can predict and differentiate both clinically normal and advanced cases from the other severity classes (peak estimations >80%) but had reduced accuracy in differentiating mild and moderate cases (peak estimations >50%). The chart, as presented, provides a simple tool to differentiate clinically normal from calves suffering the consequences of diarrhea, but fails to accurately differentiate severity for NCD related acidosis and dehydration. Further efforts are required to enhance the sensitivity and differential diagnostic value of this type of chart.

Establishing blood gas ranges in healthy bovine neonates differentiated by age, sex, and breed type.

Calf mortality and morbidity commonly occurs within the first month of life postpartum. Standard health ranges are invaluable aids in diagnostic veterinary medicine to confirm normal or the degree and nature of abnormal parameters in (sub)clinically ill animals. Extensive research has indicated significant differences between the physiologies of neonate and adult cattle, particularly for blood parameters such as pH, base excess, anion gap, and bicarbonate (HCO3-). The objective of this research was to determine the influence of age, sex, and breed type, in addition to environmental factors, on the normal blood gas profiles of neonatal calves, and thus develop a scientifically validated reference range accounting for any significant factors. The study was conducted on healthy neonatal calves (n = 288), and completed over a 2-yr period. Individual calf blood gas analysis was conducted for parameters of pH, base excess, Na+, K+, Ca2+, Cl-, glucose, total hemoglobin, HCO3-, pCO2, anion gap, strong ion difference, and hematocrit levels. Regression procedures examined the combined effect of year, farm, age, breed type, sex, and hours postfeeding on each variable. Significant effects were observed for age, sex, and breed type on several of the blood gas variables. Furthermore, year, farm, and hours postfeeding appeared to have less of an influence on neonatal bovine blood gas profiles. Consequently, specific ranges based on the neonate's age, sex, and breed type will allow for more detailed and accurate diagnosis of health and ill health in neonatal calves.

Genererating a core cluster of Fasciola hepatica virulence and immunomodulation-related genes using a comparative in silico approach.

A total of 71 virulence and immunomodulation-related transcripts (VIRs) of Fasciola hepatica have been previously proposed (Haçariz et al., 2015). In an attempt to further refine this cohort, an in silico meta analysis approach was carried out using publicly available sequence data of related liver flukes, Clonorchis sinensis and Opisthorchis viverrini. Data of both liver flukes were investigated in terms of sequential homology with data of non-parasitic organisms, pathogens and VIRs of F. hepatica, directional selection (Ka/Ks), and cytokine signaling relation (protein motif based). Some VIRs of F. hepatica [showing homology with immune receptors (for toll/interleukin-1, TGF-ß or TNF-α), TGF-ß, TNF-α, CD147, or relation with suppressors of cytokine signaling/IKBKE 1 or stimulation of TGF-ß (through thrombospondin similarity)] were found to be orthologous with those of both C. sinensis and O. viverrini. The in silico analysis indicates that on the basis of genetic commonality, a total of 30 VIRs of F. hepatica are highlighted as of foremost importance in the parasite evasion strategy, through controlling of host immune system. Findings in this study could be important to further enhance our understanding of the parasitic mechanisms and develop effective control strategies against F. hepatica and other related parasites.

The omic approach to parasitic trematode research-a review of techniques and developments within the past 5 years.

The evolution of technologies to explore parasite biology at a detailed level has made significant advances in recent years, particularly with the development of omic-based strategies. Whilst extensive efforts have been made in the past to develop therapeutic and prophylactic control strategies for trematode parasites, only the therapeutic anthelmintic approach can be regarded as usable in clinical practice. Currently, there is no commercialised prophylactic strategy (such as vaccination) for protection of the definitive host against any trematode parasite. Since 2010 in particular, the integration of omic technologies, including liquid chromatography-mass spectrometry (LC-MS) and next-generation sequencing (NGS), has been increasingly reported in trematode-related studies. Both LC-MS and NGS facilitate a better understanding of the biology of trematodes and provide a promising route to identifying clinically important biological characteristics of parasitic trematodes. In this review, we focus on the application, advantages, and disadvantages of omic technologies (LC-MS and NGS) in trematode research within the past 5 years and explore the use and translation of the omic-based research results into practical tools to deal with infection.

An observational study using blood gas analysis to assess neonatal calf diarrhea and subsequent recovery with a European Commission-compliant oral electrolyte solution.

An observational study was conducted on dairy calves (51 healthy, 31 with neonatal diarrhea) during outbreaks of diarrhea on 4 dairy farms. Clinical assessment scores (CAS) were assigned to each healthy and diarrheic calf [from 0 (healthy) to 4 (marked illness)]. Blood gas analysis [pH, base excess (BE), Na(+), K(+), Ca(2+), Cl(-), glucose, total hemoglobin, standard HCO3(-), strong ion difference (SID), and anion gap (AG)] was completed for each calf. Repeated measurements were taken in healthy animals, and pre- and postintervention measurements were taken for diarrheic calves. The mean CAS of diarrheic calves was 1.7, with 51, 30, 17, and 2% of calves scoring 1, 2, 3, and 4, respectively. The mean value for blood pH, BE, AG, and SID was 7.26, -4.93mM, 16.3mM, and 38.59mM, respectively. Calves were administered an oral rehydration and buffering solution (ORBS; Vitalife for Calves, Epsilion Ltd., Cork, Ireland) and reassessed. The mean CAS decreased to 0.38 (65% of calves scored 0 and 35% scored 1) at 6 to 18h posttreatment and to 0.03 (98% of calves scored 0 and 2% scored 1) within 24 to 48h. Significant increases in mean value for pH, BE, HCO3(-), Na(+), and SID, and significant decreases in AG, K(+), Ca(2+), and total hemoglobin were recorded posttreatment. The correlation estimates indicated that pH, HCO3(-), and BE were strongly correlated with CAS, with values exceeding 0.60 in all cases. Administration of an ORBS with a high SID and bicarbonate buffer demonstrated rapid recovery from a diarrheic episode in dairy calves.

Impact of three inactivated bovine viral diarrhoea virus vaccines on bulk milk p80 (NS3) ELISA test results in dairy herds.

Bovine viral diarrhoea virus (BVDV) is endemic in many countries and vaccines are used as a component of control and eradication strategies. Surveillance programmes to detect exposure to BVDV often incorporate the use of bulk milk (BM) testing for antibodies against BVDV p80 (NS3), but vaccination can interfere with these results. The aim of this study was to evaluate whether BVDV vaccines would confound BM testing for specific antibodies in a nationally representative group of commercial dairy farms in the Republic of Ireland. A total of 256 commercial dairy herds were included in the statistical analysis. Quarterly BM or serum samples from selected weanling heifers (unvaccinated homeborn youngstock) were assessed by ELISA for antibodies against the BVDV p80 subunit and whole virus. Wilcoxon rank-sum and receiver operating characteristic (ROC) analyses were used to examine differences among groups vaccinated with one of three commercially available inactivated BVDV vaccines. Two of the three vaccines showed evidence of interference with ELISA testing of BM samples. ROC analysis highlighted that one vaccine did not reduce the discriminatory power of the BVDV p80 ELISA for identification of herds with evidence of recent BVDV circulation, when compared with unvaccinated herds; thus, administration of this vaccine would allow uncomplicated interpretation of BM ELISA test results in vaccinated seropositive herds. Seasonal differences in BM antibody results were identified, suggesting that the latter half of lactation is the most suitable time for sampling dairy herds containing predominantly spring calving cows. The results of the present study are likely to prove useful in countries allowing vaccination during or following BVDV eradication, where BM testing is required as part of the surveillance strategy.

Generating a detailed protein profile of Fasciola hepatica during the chronic stage of infection in cattle.

Fasciola hepatica is a trematode helminth causing a damaging disease, fasciolosis, in ruminants and humans. Comprehensive proteomic studies broaden our knowledge of the parasite's protein profile, and provide new insights into the development of more effective strategies to deal with fasciolosis. The objective of this study was to generate a comprehensive profile of F. hepatica proteins expressed during the chronic stage of infection in cattle by building on previous efforts in this area. The approach included an improved sample preparation procedure for surface and internal layers of the parasite, the application of nano-UPLC-ESI-qTOF-MS (nano-ultra-performance LC and ESI quadrupole TOF MS) integrated with different acquisition methods and in silico database search against various protein databases and a transcript database including a new assembly of publically available EST. Of a total of 776 identified proteins, 206 and 332 were specific to the surface and internal layers of the parasite, respectively. Furthermore, 238 proteins were common to both layers, with comparative differences of 172 proteins detected. Specific proteins not previously identified in F. hepatica, but shown to be immunomodulatory or potential drug targets for other parasites, are discussed.

Fasciola hepatica - where is 28S ribosomal RNA?

Advanced molecular biology techniques are currently used to develop new effective strategies against fasciolosis. Assessment of the quality of extracted total RNA is an important step prior to commencing many molecular biology methods such as transcriptomics. However, RNA quality assessment is complicated for some organisms, including Fasciola hepatica, by the absence of a 28S rRNA peak/band, when assessed with modern protocols. In this study, electrophoretic profiles of F. hepatica ribosomal RNAs were evaluated using microfluidics capillary based and conventional non-denaturing gel electrophoresis methods. An important modification to recommended protocols, the exclusion of heat-denaturation step, in the microfluidics capillary based electrophoresis is critical to visualise the expected 28S rRNA and obtain an RNA integrity number (RIN). The intensity of the 28S rRNA band is reduced by the effect of non-denaturing gel electrophoresis.

A proteomic approach to investigate the distribution and abundance of surface and internal Fasciola hepatica proteins during the chronic stage of natural liver fluke infection in cattle.

Fasciola hepatica, a trematode helminth, causes an economically important disease (fasciolosis) in ruminants worldwide. Proteomic analysis of the parasite provides valuable information to understand the relationship between the parasite and its host. Previous studies have identified various parasite proteins, some of which are considered as vaccine candidates or important drug targets. However, the approximate distribution and abundance of the proteins on the surface and within internal parts of the liver fluke are unknown. In this study, two fractions including surface protein fraction (representing surface part of the parasite, near subplasma membrane of the tegument and above the basal membrane of the tegument) and internal protein fraction (representing internal part of the parasite, mainly deeper sides of the tegument including subbasal membrane and other further internal elements of the parasite) were obtained. Components of these two fractions were investigated by an advanced proteomics approach using a high-definition mass spectrometer with nano electrospray ionization source coupled to a high-performance liquid chromatography system (nanoUPLC-ESI-qTOF-MS). FABP1 was found highly abundant in the SPF fraction. Potentially novel F. hepatica proteins showing homology with AKT interacting protein (Xenopus tropicalis), sterol O-acyltransferase 2 (Homo sapiens), and integrin beta 7 (Mus musculus) were identified with high quantities in only the surface fraction of the parasite and may be possible candidates for future control strategies.

The dynamic influence of the DRB1*1101 allele on the resistance of sheep to experimental Teladorsagia circumcincta infection.

Suffolk sheep carrying the DRB1*1101 (previously referred to as-DRB1*0203 or G2) allele have been reported to show increased resistance to natural Teladorsagia circumcincta infection compared to non-carriers. The objective of this study was to compare the biochemical and physiological responses of DRB1*1101 carrier and non-carrier twin lambs to an experimental infection with 3 × 10(4) L3 Teladorsagia circumcincta. The variables studied included worm burden, faecal egg count, abomasal mast cells, IgA, IgE, IgG1 plus IgG2 and haematological parameters at 0, 3, 7, 21 and 35 days post infection (dpi), and duodenal smooth muscle contractility at 0 and 35 dpi. DRB1*1101 carrier lambs had significantly lower worm burden, higher mast cell and plasma platelet counts than the DRB1*1101 non-carriers (P < 0.05). Before infection, the non-carrier lambs exhibited significantly higher mucosal levels of all antibody isotypes measured compared to the carriers; these levels remained relatively stable over the course of infection in the non-carriers while there was a slow build up of these antibodies in the carriers up to day 21 post infection (pi). The DRB1*1101 non-carrier lambs had a significantly higher plasma lymphocyte count, and produced greater duodenal contractile force relative to the carrier lambs (P < 0.05). There was no significant difference between genotypes in the level of plasma eosinophils, monocytes, neutrophils or FEC. This evidence suggests that resistance conferred by DRB1*1101 is acquired rather than innate, depends on worm expulsion rather than fecundity and is dependent on mucosal mast cell proliferation, platelet activation, and IgA and IgE antibody responses.