Health indicators in surplus calves at the time of arrival at auction markets: Associations with distance from farms of origin in Québec, Canada.

This study evaluated the associations between estimated distance from farms' locations to auction markets, and health indicators of surplus dairy calves sold during summer 2019 and winter 2020 in Québec, Canada. A total of 3,610 animals from 1,331 different farms were used in this cross-sectional cohort study. Geographic coordinates (latitude and longitude) were obtained for each farm and the 2 participating livestock auction markets. Calves' abnormal physical signs (APS) were noted upon arrival at the auction market as they were examined by trained research staff. The haversine distance between the farm and the auction market was evaluated using geographic coordinates and categorized. Generalized linear mixed models were used for statistical analyses. The main APS observed were ocular discharge (34.9%), abnormal hide cleanliness (21.2%), swollen navel (17.2%), dehydration score 1 (at least one of the 2 following clinical signs: persistent skin tent or sunken eye, 12.9%), and dehydration score 2 (both clinical signs mentioned above, 6.5%). Calves from farms located at greater distances from the auction markets (≥110 km) had a higher risk ratio [RR = 1.08; 95% confidence internal (CI) = 1.03, 1.13] for dehydration than those from lesser distances (0-25 km). During the summertime, a RR of 1.18 (95% CI = 1.15, 1.22) was observed for dehydration compared with wintertime. A 2-way interaction between estimated distance and season showed a higher prevalence of ocular discharge for calves from farms at distances greater than or equal to 110 km during the summer (RR = 1.11; 95% CI: 1.04, 1.20) than for calves from farms located at lesser distances (0-25 km). These results demonstrate that calves from farms located at greater distances from the auction markets had more APS, mainly during the summer. A better understanding of the transport conditions and interaction with management at the farm of origin is determinant to mitigate the impact of the journey on surplus calf health.

Interplay between Bacterial Clones and Plasmids in the Spread of Antibiotic Resistance Genes in the Gut: Lessons from a Temporal Study in Veal Calves.

Intestinal carriage of extended spectrum ß-lactamase (ESBL)-producing Escherichia coli is a frequent, increasing, and worrying phenomenon, but little is known about the molecular scenario and the evolutionary forces at play. We screened 45 veal calves, known to have high prevalence of carriage, for ESBL-producing E. coli on 514 rectal swabs (one randomly selected colony per sample) collected over 6 months. We characterized the bacterial clones and plasmids carrying blaESBL genes with a combination of genotyping methods, whole genome sequencing, and conjugation assays. One hundred and seventy-three ESBL-producing E. coli isolates [blaCTX-M-1 (64.7%), blaCTX-M-14 (33.5%), or blaCTX-M-15 (1.8%)] were detected, belonging to 32 bacterial clones, mostly of phylogroup A. Calves were colonized successively by different clones with a trend in decreasing carriage. The persistence of a clone in a farm was significantly associated with the number of calves colonized. Despite a high diversity of E. coli clones and blaCTX-M-carrying plasmids, few blaCTX-M gene/plasmid/chromosomal background combinations dominated, due to (i) efficient colonization of bacterial clones and/or (ii) successful plasmid spread in various bacterial clones. The scenario "clone versus plasmid spread" depended on the farm. Thus, epistatic interactions between resistance genes, plasmids, and bacterial clones contribute to optimize fitness in specific environments. IMPORTANCE The gut microbiota is the epicenter of the emergence of resistance. Considerable amount of knowledge on the molecular mechanisms of resistance has been accumulated, but the ecological and evolutionary forces at play in nature are less studied. In this context, we performed a field work on temporal intestinal carriage of extended spectrum ß-lactamase (ESBL)-producing Escherichia coli in veal farms. Veal calves are animals with one of the highest levels of ESBL producing E. coli fecal carriage, due to early high antibiotic exposure. We were able to show that calves were colonized successively by different ESBL-producing E. coli clones, and that two main scenarios were at play in the spread of blaCTX-M genes among calves: efficient colonization of several calves by a few bacterial clones and successful plasmid spread in various bacterial clones. Such knowledge should help develop new strategies to fight the emergence of antibiotic-resistance.

Dose-dependent impact of oxytetracycline on the veal calf microbiome and resistome.

BACKGROUND: Antibiotic therapy is commonly used in animal agriculture. Antibiotics excreted by the animals can contaminate farming environments, resulting in long term exposure of animals to sub-inhibitory levels of antibiotics. Little is known on the effect of this exposure on antibiotic resistance. In this study, we aimed to investigate the long term effects of sub-inhibitory levels of antibiotics on the gut microbiota composition and resistome of veal calves in vivo. Forty-two veal calves were randomly assigned to three groups. The first group (OTC-high) received therapeutic oral dosages of 1 g oxytetracycline (OTC), twice per day, during 5 days. The second group (OTC-low) received an oral dose of OTC of 100-200 µg per day during 7 weeks, mimicking animal exposure to environmental contamination. The third group (CTR) did not receive OTC, serving as unexposed control. Antibiotic residue levels were determined over time. The temporal effects on the gut microbiota and antibiotic resistance gene abundance was analysed by metagenomic sequencing. RESULTS: In the therapeutic group, OTC levels exceeded MIC values. The low group remained at sub-inhibitory levels. The control group did not reach any significant OTC levels. 16S rRNA gene-based analysis revealed significant changes in the calf gut microbiota. Time-related changes accounted for most of the variation in the sequence data. Therapeutic application of OTC had transient effect, significantly impacting gut microbiota composition between day 0 and day 2. By metagenomic sequence analysis we identified six antibiotic resistance genes representing three gene classes (tetM, floR and mel) that differed in relative abundance between any of the intervention groups and the control. qPCR was used to validate observations made by metagenomic sequencing, revealing a peak of tetM abundance at day 28-35 in the OTC-high group. No increase in resistance genes abundance was seen in the OTC-low group. CONCLUSIONS: Under the conditions tested, sub-therapeutic administration of OTC did not result in increased tetM resistance levels as observed in the therapeutic group.

Prevalence and Transmission of Antimicrobial Resistance in a Vertically Integrated Veal Calf Production System.

Transmission of antimicrobial resistance (AMR) from animal production systems to humans through the food supply is a public health concern. Currently, little is known about the prevalence of AMR among veal calves in the United States. Therefore, the objective of this prospective cohort study was to estimate the prevalence of AMR and multidrug resistance (MDR) among Escherichia coli within a vertically integrated production system. In addition, this study aimed to identify genes associated with phenotypic resistance to third- and fourth-generation cephalosporins (3GC and 4GC). Calves from four veal cohorts were randomly sampled resulting in a total of 166 farm fecal samples, 159 harvest fecal swabs, 164 preevisceration swabs, and 122 final carcass swabs. The prevalence of MDR among random-pick E. coli isolates recovered from the respective samples was 97% (161/166), 35% (55/159), 61% (51/84), and 24% (5/21). A selective isolation protocol found cefotaxime (a 3GC)-resistant isolates in 91% (127/140) of farm fecal samples, 34% (55/164) of preevisceration swabs, and 19% (23/122) of final carcass swabs tested. Isolates resistant to cefepime, a 4GC, were found among 24% (33/140), 6.7% (11/164), and 0.8% (1/122) of the same, respective samples. Isolates resistant to ciprofloxacin, a fluoroquinolone, were recovered from 75% (73/98) of farm fecal samples, 23% (38/164) of preevisceration swabs, and 6.6% (8/122) of final carcass swabs. The blaCMY-2 and blaCTX-M resistance genes were found in 89% (93/105) and 100% (42/42) of tested subsets of 3GC- and 4GC-resistant isolates, respectively. Pulsed-field gel electrophoresis (PFGE) analysis conducted on 3GC- and fluoroquinolone-resistant isolates showed three indistinguishable PFGE patterns from cefotaxime-resistant isolates recovered at farm and from two preevisceration carcass swabs. Although the prevalence of resistance declined between initial farm fecal samples and final carcass swabs, resistant bacteria recovered from carcasses illustrate the potential transmission of AMR to the human food supply.

Impact of Clinical Salmonellosis in Veal Calves on the Recovery of Salmonella in Lymph Nodes at Harvest.

The objective of this study was to determine the prevalence, serotypes, antimicrobial resistance phenotypes, and pulsed-field gel electrophoresis (PFGE) patterns of Salmonella recovered in feces and mesenteric and prefemoral lymph nodes (LNs) from cohorts of calves with and without a confirmed outbreak of salmonellosis. In a prospective cohort study, 160 calves from four farms without a reported outbreak (nonoutbreak farms) were sampled at farm and harvest. In addition, harvest samples from 80 calves of two farms with a confirmed outbreak (outbreak farms) were collected. A culture protocol for Salmonella isolation was applied for all samples and recovered isolates were further characterized by serotyping, antimicrobial susceptibility testing, and PFGE. Among nonoutbreak farms, Salmonella was recovered from 0% (0/160) farm fecal samples, 3.7% (6/160) harvest fecal swabs, 21.9% (35/160) mesenteric LNs, and 0.6% (1/160) prefemoral LNs. Serotypes identified in nonoutbreak herds included Salmonella Typhimurium, Cerro, Hartford, and Newport. Most isolates (64.3%, 27/42) exhibited a unique multidrug-resistant (MDR) phenotype, including resistance to extended-spectrum cephalosporins. Salmonella prevalence in harvest fecal samples and prefemoral LNs among calves from outbreak farms was numerically higher, but not significantly different than those without an outbreak. Serotypes recovered from outbreak farms included Salmonella Heidelberg and Typhimurium, and the monophasic Salmonella Typhimurium strains 4,5,12:i:- and 4,12:i:-, which have been also reported as highly pathogenic in humans. All isolates (33/33) exhibited an MDR phenotype. Salmonella strains recovered from ill calves in two outbreaks had indistinguishable PFGE patterns, suggesting between-farm transmission. In addition, the genotype of Salmonella Heidelberg causing an outbreak among calves was recovered from three prefemoral LNs of surviving members of the cohort at harvest. Implementation of preharvest biosecurity measures (limited personnel and visitor traffic, vehicle, footwear, and utensils disinfection) should be highly recommended to decrease the prevalence of Salmonella on farms and safeguard the food safety.

Substantial replacement of lactose with fat in a high-lactose milk replacer diet increases liver fat accumulation but does not affect insulin sensitivity in veal calves.

In veal calves, the major portion of digestible energy intake originates from milk replacer (MR), with lactose and fat contributing approximately 45 and 35%, respectively. In veal calves older than 4 mo, prolonged high intakes of MR may lead to problems with glucose homeostasis and insulin sensitivity, ultimately resulting in sustained insulin resistance, hepatic steatosis, and impaired animal performance. The contribution of each of the dietary energy sources (lactose and fat) to deteriorated glucose homeostasis and insulin resistance is currently unknown. Therefore, an experiment was designed to compare the effects of a high-lactose and a high-fat MR on glucose homeostasis and insulin sensitivity in veal calves. Sixteen male Holstein-Friesian calves (120±2.8kg of BW) were assigned to either a high-lactose (HL) or a high-fat (HF) MR for 13 consecutive weeks. After at least 7 wk of adaptation, whole-body insulin sensitivity and insulin secretion were assessed by euglycemic-hyperinsulinemic and hyperglycemic clamps, respectively. Postprandial blood samples were collected to assess glucose, insulin, and triglyceride responses to feeding, and 24-h urine was collected to quantify urinary glucose excretion. At the end of the trial, liver and muscle biopsies were taken to assess triglyceride contents in these tissues. Long-term exposure of calves to HF or HL MR did not affect whole-body insulin sensitivity (averaging 4.2±0.5×10-2 [(mg/kg∙min)/(µU/mL)]) and insulin secretion. Responses to feeding were greater for plasma glucose and tended to be greater for plasma insulin in HL calves than in HF calves. Urinary glucose excretion was substantially higher in HL calves (75±13g/d) than in HF calves (21±6g/d). Muscle triglyceride content was not affected by treatment and averaged 4.5±0.6g/kg, but liver triglyceride content was higher in HF calves (16.4±0.9g/kg) than in HL calves (11.2±0.7g/kg), indicating increased hepatic fat accumulation. We conclude that increasing the contribution of fat to the digestible energy intake from the MR from 20 to 50%, at the expense of lactose does not affect whole-body insulin sensitivity and insulin secretion in calves. However, a high-lactose MR increases postprandial glucose and insulin responses, whereas a high-fat MR increases fat accumulation in liver but not muscle.

Lactose in milk replacer can partly be replaced by glucose, fructose, or glycerol without affecting insulin sensitivity in veal calves.

Calf milk replacer (MR) contains 40 to 50% lactose. Lactose strongly fluctuates in price and alternatives are desired. Also, problems with glucose homeostasis and insulin sensitivity (i.e., high incidence of hyperglycemia and hyperinsulinemia) have been described for heavy veal calves (body weight >100 kg). Replacement of lactose by other dietary substrates can be economically attractive, and may also positively (or negatively) affect the risk of developing problems with glucose metabolism. An experiment was designed to study the effects of replacing one third of the dietary lactose by glucose, fructose, or glycerol on glucose homeostasis and insulin sensitivity in veal calves. Forty male Holstein-Friesian (body weight=114 ± 2.4 kg; age=97 ± 1.4 d) calves were fed an MR containing 462 g of lactose/kg (CON), or an MR in which 150 g of lactose/kg of MR was replaced by glucose (GLU), fructose (FRU), or glycerol (GLY). During the first 10d of the trial, all calves received CON. The CON group remained on this diet and the other groups received their experimental diets for a period of 8 wk. Measurements were conducted during the first (baseline) and last week of the trial. A frequently sampled intravenous glucose tolerance test was performed to assess insulin sensitivity and 24 h of urine was collected to measure glucose excretion. During the last week of the trial, a bolus of 1.5 g of [U-(13)C] substrates was added to their respective meals and plasma glucose, insulin, and (13)C-glucose responses were measured. Insulin sensitivity was low at the start of the trial and remained low [1.2 ± 0.1 and 1.0 ± 0.1 (mU/L)(-1) × min(-1)], and no treatment effect was noted. Glucose excretion was low at the start of the trial (3.4 ± 1.0 g/d), but increased in CON and GLU calves (26.9 ± 3.9 and 43.0 ± 10.6g/d) but not in FRU and GLY calves. Postprandial glucose was higher in GLU, lower in FRU, and similar in GLY compared with CON calves. Postprandial insulin was lower in FRU and GLY and similar in GLU compared with CON calves. Postprandial (13)C-glucose increased substantially in FRU and GLY calves, indicating that calves are able to partially convert these substrates to glucose. We concluded that replacing one third of lactose in MR by glucose, fructose, or glycerol in MR differentially influences postprandial glucose homeostasis but does not affect insulin sensitivity in veal calves.

Surveillance of Antimicrobial Resistance of Escherichia coli Isolates from Intestinal Contents of Dairy and Veal Calves in the Veneto Region, Northeaster Italy.

This surveillance study aimed to estimate the proportion of antimicrobial resistant strains and antimicrobial resistance (AMR) profiles of E. coli isolates detected from the intestinal contents of veal and dairy calves in the Veneto Region, Northeaster Italy. Additionally, we investigated the differences in AMR profiles between dairy and veal calves over the period 2017-2022. Overall 1150 E. coli isolates were tested from calves exhibiting enteric disease, with 868 from dairy and 282 from veal calves. The percentage of resistant isolates to nine antimicrobials was notably higher in veal calves compared to dairy calves, except for ampicillin. Throughout the study period, we observed a significant increase in the proportion of resistant isolates to florfenicol, gentamycin, paromomycin, tetracycline and trimethoprim/sulfamethoxazole in dairy calves, while we did not detect any significant increase in the proportion of resistant isolates among veal calves. A substantial proportion (75.9%) of the isolated E. coli exhibited multi-drug resistance (MDR). The proportion of multi-drug resistant isolates was significantly higher in veal calves (91.7%) compared to dairy calves (74.3%) all through the surveillance period (2017-2022), with no significant variation in MDR proportion among veal calves between 2017 and 2022 but a significant increase among dairy calves.

Inter-Rater Reliability of Scoring Systems for Abomasal Lesions in Quebec Veal Calves.

The objective of this study was to determine the inter-rater reliability of current scoring systems used to detect abomasal lesions in veal calves. In addition, macroscopic lesions were compared with corresponding histological lesions. For this, 76 abomasa were retrieved from veal calves in a slaughterhouse in Quebec and scored by four independent raters using current scoring systems. The localisations of the lesions were separated into pyloric, fundic, or torus pyloricus areas. Lesions were classified into three different types, i.e., erosions, ulcers, and scars. To estimate the inter-rater reliability, the coefficient type 1 of Gwet's agreement and Fleiss κ were used for the presence or absence of a lesion, and the intra-class correlation coefficient was used for the number of lesions. All veal calves had at least one abomasal lesion detected. Most lesions were erosions, and most of them were located in the pyloric area. Overall, a poor to very good inter-rater agreement was seen for the pyloric area and the torus pyloricus regarding the presence or absence of a lesion (Fleiss κ: 0.00-0.34; Gwet's AC1: 0.12-0.83), although a higher agreement was observed when combining all lesions in the pyloric area (Fleiss κ: 0.09-0.12; Gwet's AC1: 0.43-0.93). For the fundic area, a poor to very good agreement was also observed (Fleiss κ: 0.17-0.70; Gwet's AC1: 0.90-0.97). Regarding the inter-rater agreement for the number of lesions, a poor to moderate agreement was found (ICC: 0.11-0.73). When using the scoring system developed in the European Welfare Quality Protocol, a poor single random rater agreement (ICC: 0.42; 95% CI: 0.31-0.56) but acceptable average random rater agreement (ICC: 0.75; 95% CI: 0.64-0.83) was determined. Microscopic scar lesions were often mistaken as ulcers macroscopically. These results show that the scoring of abomasal lesions is challenging and highlight the need for a reliable scoring system. A fast, simple, and reliable scoring system would allow for large scale studies which investigate possible risk factors and hopefully help to prevent these lesions, which can compromise veal calves' health and welfare.

Diagnostic and Prognostic Value of Clinical Scoring and Lung Ultrasonography to Assess Pulmonary Lesions in Veal Calves.

This study on veal calf respiratory disease assessed the association between an on-farm clinical scoring system and lung ultrasonography with the postmortem inspection of the lungs. The comparisons allowed the calculation of predictive values of the diagnostic methods. In total, 600 calves on an Austrian veal calf farm were examined at the beginning and the end of the fattening period. Overall, the area under the curve (AUC) for ultrasonographic scores was 0.90 (rsp = 0.78) with a sensitivity (Se) of 0.86. The specificity (Sp) was 0.78, and the positive predictive value (PPV) was 0.74. The AUC for the physical examination was 0.76 (rsp = 0.55) with a Se of 0.64, an Sp of 0.81, and a PPV of 0.69. For the combination of ultrasonography and physical examination, an AUC curve of 0.85 (rsp = 0.69) was calculated. A Se of 0.65 and a Sp of 0.88 with a PPV of 0.73 was calculated. This study concluded that both physical and ultrasonographic examination scoring are reliable examination methods for the detection of lung diseases in veal calves.

Oxazolidinone resistance genes in florfenicol-resistant enterococci from beef cattle and veal calves at slaughter.

Background: Linezolid is a critically important oxazolidinone antibiotic used in human medicine. Although linezolid is not licensed for use in food-producing animals, the use of florfenicol in veterinary medicine co-selects for oxazolidinone resistance genes. Objective: This study aimed to assess the occurrence of cfr, optrA, and poxtA in florfenicol-resistant isolates from beef cattle and veal calves from different herds in Switzerland. Methods: A total of 618 cecal samples taken from beef cattle and veal calves at slaughter originating from 199 herds were cultured after an enrichment step on a selective medium containing 10 mg/L florfenicol. Isolates were screened by PCR for cfr, optrA, and poxtA which are genes known to confer resistance to oxazolidinones and phenicols. One isolate per PCR-positive species and herd was selected for antimicrobial susceptibility testing (AST) and whole-genome sequencing (WGS). Results: Overall, 105 florfenicol-resistant isolates were obtained from 99 (16%) of the samples, corresponding to 4% of the beef cattle herds and 24% of the veal calf herds. Screening by PCR revealed the presence of optrA in 95 (90%) and poxtA in 22 (21%) of the isolates. None of the isolates contained cfr. Isolates included for AST and WGS analysis were Enterococcus (E.) faecalis (n = 14), E. faecium (n = 12), E. dispar (n = 1), E. durans (n = 2), E. gallinarum (n = 1), Vagococcus (V.) lutrae (n = 2), Aerococcus (A.) urinaeequi (n = 1), and Companilactobacillus (C.) farciminis (n = 1). Thirteen isolates exhibited phenotypic linezolid resistance. Three novel OptrA variants were identified. Multilocus sequence typing identified four E. faecium ST18 belonging to hospital-associated clade A1. There was a difference in the replicon profile among optrA- and poxtA-harboring plasmids, with rep9 (RepA_N) plasmids dominating in optrA-harboring E. faecalis and rep2 (Inc18) and rep29 (Rep_3) plasmids in poxtA-carrying E. faecium. Conclusion: Beef cattle and veal calves are reservoirs for enterococci with acquired linezolid resistance genes optrA and poxtA. The presence of E. faecium ST18 highlights the zoonotic potential of some bovine isolates. The dispersal of clinically relevant oxazolidinone resistance genes throughout a wide variety of species including Enterococcus spp., V. lutrae, A. urinaeequi, and the probiotic C. farciminis in food-producing animals is a public health concern.

Development of an equation to screen for solar hemorrhages from digital cushion ultrasound texture analysis in veal calves at slaughter.

Claw disorders are a relevant welfare issue in the cattle industry, fast and accurate diagnoses are essential for successful treatment and prevention. The present study aimed to develop an equation to assess the presence of solar hemorrhages from real-time ultrasound images texture analysis at slaughter. Eighty-eight hind feet were collected at the slaughterhouse from 44 Holstein male veal calves. The claws were trimmed by a veterinarian hoof-trimmer, approximately 30 min after the calves' slaughter, and classified into healthy and affected by solar hemorrhages. At the same time, ultrasound images were collected for each claw. Sole soft tissues' thickness was measured, and texture analysis was performed using MaZda software. The resulting parameters from sole soft tissues' measurements and texture analysis were screened with a stepwise linear discriminant analysis using the absence or presence (0/1) of solar hemorrhages as the dependent variable. Results from the stepwise analysis identified 9 variables (among 279) as predictors, and an equation was developed and used to predict the presence or absence of solar hemorrhages on the scanned claws by binary measure: values ≤0.5 counted as 0, while those >0.5 as 1. Validation of the equation was performed by testing predicted lesions (LESpred) against the clinically evaluated lesions (LESeval) with a confusion matrix, a ROC analysis, and a precision-recall curve. Results of the present study suggest that the equation proposed has a good potential for detecting effectively hemorrhages of the sole by ultrasound imaging texture means, and could be used to monitor unsatisfactory housing and management conditions at the farm level, and for early management intervention and prevention.

Predicting veal-calf trading events in France.

Global trade has been ranked as one of the top five drivers of infectious disease threat events. More specifically, livestock trade is known to increase the speed at which infectious diseases circulate and to facilitate their dissemination over large distances Therefore, predicting animal movements arising from trade is crucial for assessing epidemic risk and the impact of preventive measures. In this study, we developed a statistical framework for predicting trading events using predictors accessible from routinely collected data. We focused on veal calves, a category of animals with significant commercial value; the dataset considered the veal calf trade in France between January 2011 and June 2019. A subset of farms with consistent trade behaviour over time was built to be used throughout the study. To predict sale or purchase event occurrences, our predictive framework was based on random forests as a binary classification tool, an approach that allows a large number of potential predictors. We explored the robustness of model predictions with respect to the delay in data acquisition and prediction lag time. Overall, sales were more accurately predicted than purchasing events. Unsurprisingly, a delay in data acquisition led to a decrease in the performance of indicators, whereas prediction lag time had little impact. Sale-related predictors mostly reflected past trading events, whereas purchase-related predictors were associated with past trading events, farm management and general farm characteristics. The model outputs also suggested that the veal calf trading network is driven by sales rather than by purchases. Regardless of the length of the delay in data acquisition and prediction lag, the random forest approach fitted on data with municipality as trading unit and a 28-day trading period provided better performance scores (F1-score, positive predictive value and negative predictive value) than scenarios with finer temporal and spatial aggregation units. Predicted trade events can therefore be used to reconstruct the entire veal calf trading network and transfers between selling and purchasing units for each period. This predicted network could be further used to simulate the spread of pathogens via animal trade.

Effect of Dietary Hemp Cake Inclusion on the In Vivo and Post Mortem Performances of Holstein Veal Calves.

Fifty-two male Holstein veal calves were divided into two homogeneous groups receiving two isoenergetic and isonitrogenous concentrates without (CTR group) or with 3% of hemp cake (HC group). The trial lasted for 171 days. All the calves were weighed five times during the trial. At slaughtering, carcasses were weighed and measured. Meat quality was determined on the Longissimus dorsi muscle. Average daily gain in the first period of the experiment (0-80 d) and dressing percentage and rump width of the carcasses were higher in HC group. Cooking weight losses and shear force were higher in the meat of the HC group while color parameters were similar in the two experimental groups. Unexpectedly, the alpha-linolenic acid content of meat was lower in the HC group. In conclusion, hemp cake can be considered an interesting ingredient in the concentrate used for the production of veal calves, but further studies will be needed to determine a suitable dosage in order to improve the nutritional quality of meat (i.e., the n-3 fatty acids content) without negative effects on physical characteristics.

Respiratory pathogens in veal calves: Inventory of circulating pathogens.

In the veal industry in The Netherlands, each year around 1.2 million "white" veal calves are produced on around 1100 farms. Bovine respiratory disease (BRD) causes serious health issues in these calves, also resulting in high usage of antimicrobials. To reduce antimicrobial usage, a more targeted treatment regime is needed, for which it is necessary to identify the causative agent. This study aimed at determining associations between pathogens and clinical disease, between prevalence of pathogens and BRD outbreaks, and BRD and performance. A cohort study was conducted involving ten veal farms, in which calf respiratory health was evaluated for the first 12 weeks. Whenever there was an outbreak of BRD, as determined by the farm veterinary surgeon, samples were taken from diseased and control calves through broncho-alveolar lavage. From these samples a broad spectrum of micro-organisms were isolated. Performance data were also collected. A total of 23 outbreaks happened during the 12 week study period, mostly in the first six weeks. BRD associated pathogens found were: BHV1, BPI3V, BRSV, BVDV, Pasteurella multocida, Mannheimia haemolytica, Trueperella pyogenes, Histophilus somni, Mycoplasma bovis, Mycoplasma bovirhinis and Mycoplasma dispar. For most BRD associated pathogens, there was no clear association between presence or prevalence of the micro-organisms and clinical issues. Only T. pyogenes (7.4% in healthy, 14.6% in diseased calves, p 0.013), M. bovis (37.6% and 63.2% respectively, p 0.001) and BVDV (9.9% and 16.9% respectively, p 0.03) were found more often in diseased animals. BPI3V was found in a few early outbreaks, which might suggest involvement in early outbreaks. It appears to be difficult to associate specific pathogens to outbreaks at the species level. BRD is the major reason for treatment with antimicrobials. More specific knowledge about the association between pathogens and health/disease could help to reduce antimicrobial use.

Longitudinal study on the prevalence of extended spectrum cephalosporins-resistant Escherichia coli colonization in Dutch veal farms.

A longitudinal study was performed to investigate the prevalence of Extended-Spectrum Cephalosporin-Resistant (ESC-R) Escherichia coli colonization in Dutch veal farms. Rectal swabs from 683 calves born in 13 Dutch dairy farms were collected one day prior to transportation to the veal farm at 14 or 28 days of age, and at 5 different time points 8 Dutch veal farms. In addition, characteristics of the calf, cows, and farm management were collected. Rectal swabs were selectively cultured for ESC-R E. coli. In total, 1202 ESC-R E. coli isolates were recovered. Overall, the prevalence of ESC-R E. coli increased from 24.4 % at one day prior to transportation to 57.3 % in week two after arrival of calves at the veal farm. No associations were found between the presence of ESC-R E. coli at the dairy or veal farm and age of transportation, sex and breed. The presence of ESC-R E. coli in week 6, 10, and 18 at the veal farm was positively associated with the presence of ESC-R E. coli in week 10, 18, and 24, respectively (p < 0.05). Individual antibiotic treatments applied before week 2 and 6 upon arrival to the veal farms tended to increase the ESC-R E. coli colonization frequency. Our results indicate that ESC-R E. coli colonization frequency substantially increases after arrival of calves on the veal farm. In addition to individual antibiotic treatments, it is considered likely that frequently applied batch antibiotic treatments are also implicated in the ESC-R E. coli colonization frequency.

Validation of serum paraoxonase/arylesterase 1 (PON1) as a protein marker of illicit dexamethasone treatment in veal calves.

The illicit use of dexamethasone and other glucocorticoids for cattle fattening in livestock production has been widely described; evidence for illegal treatments can be obtained by direct or indirect detection. In our previous study, we applied two-dimensional electrophoresis (2DE) to identify plasma protein markers of dexamethasone administration in veal calves. Comparison of 2DE maps obtained from blood samples before and after treatment showed the disappearance of two protein spots identified as serum paraoxonase/arylesterase 1 precursor (PON1). In the present study, we validated PON1 as a marker by analysing a larger number of samples treated with dexamethasone for illicit use. Analysis of samples from experimental treatment with other glucocorticoids, androgens and oestrogens confirmed that their influence on PON1 could be excluded. The specificity of the PON1 protein marker was verified on expected negative field samples to exclude interfering factors. However, there is poor statistical evidence to support a significant association between the outcome of PON1 and the considered variables. The results on field samples were compared with histological examination of the thymus as a biomarker of corticosteroid treatment monitored in the Italian histological plan for the control of growth promoters in animals. Two suspect cases were identified from two Piedmont farms where other animals had tested positive at histological examination. In conclusion, the absence of PON1 in the plasma of veal calves can indirectly reveal illicit dexamethasone treatment in individual animals and so identify suspect farms for further investigation. It is effective in a period ranging from 3 to about 10 days from illicit treatment, covering a time span that goes beyond the limits of official chemical controls and preceding histological controls on the thymus of slaughtered animals. PON1 detection in plasma can be coupled with other tests to identify illegal dexamethasone use on veal calf farms.

High Fecal Prevalence of mcr-Positive Escherichia coli in Veal Calves at Slaughter in France.

The aim of this study was to determine the percentage of healthy veal calves carrying mcr-positive E. coli strains at the time of slaughter in France. Fecal samples were selectively screened for mcr-positive E. coli isolates using media supplemented with colistin. Screening for mcr genes was also carried out in E. coli isolates resistant to critically important antimicrobials used in human medicine recovered from the same fecal samples. Overall, 28 (16.5%) out of the 170 veal calves tested carried mcr-positive E. coli. As some calves carried several non-redundant mcr-positive strains, 41 mcr-positive E. coli were recovered. Thirty-one and seven strains were positive for mcr-1 and mcr-3 genes, respectively, while no strain was positive for the mcr-2 gene. Co-carriage of mcr-1 and mcr-3 was identified in three strains. All mcr-positive E. coli isolates, except one, were multidrug-resistant, with 56.1% being ciprofloxacin-resistant and 31.7% harboring blaCTX-M genes. All mcr-3-positive E. coli carried blaCTX-M genes, mainly blaCTX-M-55. This study highlights the high prevalence of mcr-positive E. coli strains in feces of veal calves at the time of slaughter. It also points out the multidrug (including ciprofloxacin) resistance of such strains and the co-occurrence of mcr-3 genes with blaCTX-M-55 genes.

Changes in Fecal Carriage of Extended-Spectrum ß-Lactamase Producing Enterobacterales in Dutch Veal Calves by Clonal Spread of Klebsiella pneumoniae.

This study aimed to characterize the changes in fecal carriage of Extended-Spectrum ß-Lactamase (ESBL) producing Enterobacterales (ESBL-PE) in a single Dutch veal calves. During the rearing period at the Dutch veal farm, a decrease in fecal carriage of cefotaxime-resistant Escherichia coli isolates was observed after 2 weeks at the veal farm, while an increase of cefotaxime-resistant Klebsiella pneumoniae isolates was demonstrated. E. coli and K. pneumoniae were isolated from rectal swabs collected from 110 veal calves in week 2, 6, 10, 18, and 24 after their arrival at the farm. ESBL-PE isolates were selectively cultured and identified by MALDI-TOF. ESBL genes were characterized by RT-PCR, PCRs, and amplicon sequencing. A total of 80 E. coli and 174 K. pneumoniae strains were isolated from 104 out of 110 veal calves. The prevalence of ESBL-E. coli decreased from week 2 (61%) to week 6 (7%), while an unexpected increase in ESBL-K. pneumoniae colonization was detected in week 6 (80%). The predominant ESBL genes detected in E. coli isolates were bla CTX-M-15 and the non-ESBL gene bla TEM-1a, while in K. pneumoniae bla CTX-M-14 gene was detected in all isolates. Four cefotaxime-resistant K. pneumoniae isolates were randomly selected and characterized in deep by transformation, PCR-based replicon typing, and whole-genome sequencing (WGS). The clonal relatedness of a subgroup of nine animals carrying K. pneumoniae ESBL genes was investigated by Multi Locus sequence typing (MLST). In four ESBL-K. pneumoniae isolates, bla CTX-M-14 was located on IncFIIK and IncFIINK plasmid replicons and the isolates were multi-drug resistant (MDR). MLST demonstrated a clonal spread of ESBL-K. pneumoniae ST107. To the best of our knowledge, this is the first study to report a change in fecal carriage of ESBL-PE over time in the same veal calf during the rearing period.

Transcriptional Biomarkers and Immunohistochemistry for Detection of Illicit Dexamethasone Administration in Veal Calves.

Corticosteroids such as Dexamethasone (DEX) are commonly licensed for therapy in meat animals due to their known pharmacological properties. However, their misuse aimed to achieve anabolic effects is often found by National Residues Control Plans. The setup of a complementary "biomarker based" methods to unveil such illicit practices is encouraged by current European legislation. In this study, the combined use of molecular and histological quantitative techniques was applied on formalin fixed paraffin embedded (FFPE) muscle samples to assess the effects of illicit DEX treatment on veal calves. A PCR array, including 28 transcriptional biomarkers related to DEX exposure, was combined with a histochemical analysis of muscle fiber. An analysis based on unsupervised (PCA) and supervised (PLS-DA and Kohonen's SOM) methods, was applied in order to define multivariate models able to classify animals suspected of illicit treatment by DEX. According to the conventional univariate approach, a not-significant reduction in type I fibres was recorded in the DEX-treated group, and only 12 out of 28 targeted genes maintained their expected differential expression, confirming the technical limitations of a quantitative analysis on FFPE samples. However, the multivariate models developed highlighted the possibility to establish complementary screening strategies, particularly when based on transcriptional biomarkers characterised by low expression profiles.