Training Laboratory Rabbits to Refine Routine Husbandry Procedures.

Non-aversive handling and training techniques for laboratory animals are required to facilitate experimental and routine husbandry procedures, improving both animal welfare and scientific quality. Clicker training was utilized to develop training protocols for rabbits to refine stressful routine husbandry procedures usually associated with lifting (i.e., being picked up from the floor)/restraining (i.e., being held in the arms of a human) them. Thirteen female New Zealand White rabbits were trained over three weeks. All rabbits learned the predefined goal behaviors: they followed the target stick, jumped onto the weighing scale, entered a transport box, and reared while placing their front paws onto the trainer's hand. In addition, ten animals jumped from the floor onto the sitting trainer's lap and allowed the trainer to lift their paws off the surface while sitting on the trainer's lap. For some individuals, the protocols had to be adapted by additional interim steps. At the end of the training, the rabbits reliably showed the expected goal behaviors, even after short and long training breaks. With few exceptions, a familiar person other than the trainer could elicit the goal behaviors from the rabbits (generalization), though further sessions were required for generalization. In the voluntary approach test, the rabbits preferred interacting with the trainer in the 1st trial but spent as much time with an unfamiliar person as with the trainer in the 2nd trial. The behavioral observations suggested that picking the rabbits up with the transport box, as described in the protocol, instead of restraining them with the scruff of their neck and lifting them on the arm, was less aversive. All in all, the training protocols were feasible and can serve as a refinement strategy in laboratory animal facilities. In the interest of animal welfare, the training protocols should be applied wherever possible.

Academics meet practitioners: Interactive exchange instead of consuming knowledge on dairy calving management.

Consensus on optimal calving management is difficult to achieve among veterinarians. Some of this discordance may stem from asymmetry between the literature written by subject matter experts (SME) and the knowledge, attitude and practice (KAP) of non-SMEs and veterinary practitioners. Hence, the objective of this study was to assess how veterinary practitioners KAPs in calving management compared with the findings in the literature. An online questionnaire survey was conducted prior to a World Buiatrics Congress by delegates (n = 80) who attended a workshop on calving management at which the outcomes of the latter were recorded. The responses from the questionnaire and the consensus from the workshop were compared and these were compared with the literature on calving management, specifically, prediction of calving, assistance at calving and farmer training in calving management. In general, there was good agreement between the aggregated individual responses to the questionnaire and the consensuses from the workshop. The suggested methods of predicting calving were well aligned between practitioners and those researched in the literature but practitioners tended to emphasize more practical aspects of prediction while the literature tends to focus more on the effectiveness of various predictor devices. Practitioners tended to recommend a later time of calving assistance than that recommended in the recent literature though they did also emphasize the importance of calving progress. The majority of practitioners conducted informal (at calving) calving training focusing on time of intervention, welfare-friendly intervention and delivery of a live healthy calf. However, the educational literature recommends a formal training approach as being more effective. These findings indicate that practitioners may benefit from continuing professional education on the scientific merits/demerits of calving prediction devices, current best practice on intervention timing during calving and formal methods of farmer training on calving management.

Residue Concentrations of Cloxacillin in Milk after Intramammary Dry Cow Treatment Considering Dry Period Length.

Dry cow treatment with an intramammary antibiotic is recommended to reduce the risk of mastitis at the beginning of the next lactation. The dry period may be shortened unintentionally, affecting antibiotic residue depletion and the time when residues reach concentrations below the maximum residue limit (MRL). The objective of this study was to evaluate residue depletion in milk after dry cow treatment with cloxacillin, considering dry periods of 14 (G14d), 21 (G21d), and 28 d (G28d). Overall, fifteen cows with 60 udder quarters were included in the study. For each cow, three of the udder quarters were treated with 1000 mg cloxacillin benzathine (2:1) on d 252, d 259, and d 266 of gestation; one quarter was left untreated. Milk samples were drawn until 20 DIM and milk composition, somatic cell count and cloxacillin residues were analyzed. The HPLC-MS/MS revealed different excretion kinetics for the compounds cloxacillin and cloxacillin benzathine (1:1). All cows showed a cloxacillin and cloxacillin benzathine (1:1) concentration below the MRL of 30 µg/kg after 5 d. In the udder quarters of G21d and G28d, the cloxacillin concentration was already below the MRL at first milking after calving. The cloxacillin benzathine (1:1) concentration in the milk of G28d, G21d, and G14d fell below 30 µg/kg on the 5th, 3rd, and 5th DIM, respectively. Shortening the dry period affects residue depletion after dry cow treatment with cloxacillin. The risk of exceeding the MRL, however, seems low, even with dry periods shorter than 14 d.

Detection of Hypoglycin A and MCPrG Metabolites in the Milk and Urine of Pasture Dairy Cows after Intake of Sycamore Seedlings.

Hypoglycin A (HGA), methylenecyclopropylglycine (MCPrG), hypoglycin B (HGB), and γ-glutamyl-α-(methylenecyclopropyl) glycine (γ-glutamyl-MCPrG) are secondary plant metabolites occurring in sycamore maple (Acer pseudoplatanus) as well as several other Sapindaceae (e.g., Blighia sapida). By interfering with energy metabolism, they may cause severe intoxication in humans and other species. However, to date, there is not enough data available concerning the intake, metabolism, or excretion of sycamore maple toxins in dairy cows. In May 2022, five cows were observed over four days, when they had first access to a pasture with two sycamore maples. Grazing of their seedlings that grew numerously in between the pasture plants was monitored by direct observation. Milk samples were drawn both from individual cows and from the bulk tank. Spontaneous urine samples were collected from all cows on day 3 after access to the pasture. Seedlings (100 g) were sampled on the pasture and analyzed, together with milk and urine samples, for sycamore toxins and their metabolites using liquid chromatography-tandem mass spectrometry and liquid chromatography-high-resolution mass spectrometry. Cows ingested sycamore seedlings while grazing. Values of HGA in milk were below the limit of quantification. However, metabolites of HGA and MCPrG were detected in individual milk samples already at the end of the first day of grazing. Urine samples of all five cows showed higher concentrations of conjugated HGA and MCPrG metabolites than in milk. Observations suggest that dairy cows may have a low susceptibility toward sycamore maple toxins. However, whether this could be attributed to foregut fermenting species in general requires further elucidation.

[Body temperature measurement in pigs: Are infrared thermometers a non-invasive alternative?] / Messung der Körpertemperatur beim Schwein: Können Infrarotthermometer eine Alternative sein?

OBJECTIVE: Internal body temperature is an essential parameter in evaluation an animal's general health status. The rectal temperature as 'gold standard' requires restraining of the animal which may cause stress especially when not accustomed to handling procedures. Stress on the other hand should whenever possible be avoided as it negatively affects animal welfare and may increase body temperature. The present study evaluated whether measuring the body surface temperature with an infrared thermometer (IRT) may represent a stressless alternative method to rectal body temperature measurements. MATERIAL AND METHODS: Twelve male fattening pigs were included in the study. Body temperature was measured once a week for 11 weeks. Body surface temperature measurements were performed in the areas of the forehead, caudal base of the ear and anus using two infrared thermometers (IRT1 and IRT2) each. RESULTS: Throughout the study, all pigs were clinically healthy. Best repeatability was found for the rectal thermometer and IRT1 in the anus region. Homogeneity of variance was not found for the measurements of the three thermometers. Mean values of body temperature were significantly different (p<0,05) between thermometers and measurement points. Thereby, the type of thermometer and measurement point possessed a moderate to strong effect. The Bland-Altman plot shows that differences in the values of the thermometers and measurement points are within the acceptable range of variation (95% interval). However, the range of variation is too substantial for clinical assessment of the body temperature. CONCLUSION: The repeatability of temperature data measured with IRT on the body surface of pigs is acceptable. For this procedure, restraining the animals is not necessary, therefore reducing the animal's stress level during the clinical examination. However, the correlation to the rectal body temperature is weak to moderate. CLINICAL RELEVANCE: In order to use IRT for health monitoring in animals, reference values for respective IRT and measurement points need to be established. In the current study no case of hyper- or hypothermia occurred. Further research is warrented to evaluate whether IRT reliably detect fever.

Longitudinal Study for the Detection and Quantification of Campylobacter spp. in Dairy Cows during Milking and in the Dairy Farm Environment.

Campylobacteriosis outbreaks have repeatedly been associated with the consumption of raw milk. This study aimed to explore the variation in the prevalence and concentration of Campylobacter spp. in cows' milk and feces, the farm environment and on the teat skin over an entire year at a small German dairy farm. Bi-weekly samples were collected from the environment (boot socks), teats, raw milk, milk filters, milking clusters and feces collected from the recta of dairy cows. Samples were analyzed for Campylobacter spp., E. coli, the total aerobic plate count and for Pseudomonas spp. The prevalence of Campylobacter spp. was found to be the highest in feces (77.1%), completely absent in milking equipment and low in raw milk (0.4%). The mean concentration of Campylobacter spp. was 2.43 log10 colony-forming units (CFU)/g in feces and 1.26 log10 CFU/teat swab. Only a single milk filter at the end of the milk pipeline and one individual cow's raw milk sample were positive on the same day, with a concentration of 2.74 log10 CFU/filter and 2.37 log10 CFU/mL for the raw milk. On the same day, nine teat swab samples tested positive for Campylobacter spp. This study highlights the persistence of Campylobacter spp. for at least one year in the intestine of individual cows and within the general farm environment and demonstrates that fecal cross-contamination of the teats can occur even when the contamination of raw milk is a rare event.

Why workshops work: Examining the efficacy of training trainers to train goats.

Experimental procedures involving farm animals are often associated with stress due to restraining. Stress can be reduced through use of positive reinforcement training, which then serves as refinement according to the 3Rs principles. Trainer skills, however, may influence the feasibility and success of animal training. The potential influence of trainer skills as well as the education of animal trainers are rarely described in literature but are necessary information for the implementation of positive reinforcement training as a refinement measure. To investigate the effect of educational programmes on animal trainers, we compared the training success of two groups of participants in training goats to elicit a behaviour that would allow simulated venipuncture. One group was educated in a two-day workshop while the other was provided with specific literature for self-instructed learning. Training success was evaluated using an assessment protocol developed for this study. A greater training success in the WORKSHOP GROUP, reflected by objective and subjective measures, was clearly supported statistically. In addition, 73 versus only 13% of the participants of the WORKSHOP GROUP and the self-instructed BOOK GROUP, respectively, stated that they could completely implement the knowledge gained in the course of this study. Our results indicate that more intensively educated trainers can train animals more successfully. In conclusion, if animal training is implemented as refinement, animal caretakers should receive instruction for positive reinforcement training.

"Do not look at me like that": Is the facial expression score reliable and accurate to evaluate pain in large domestic animals? A systematic review.

Introduction: Facial expression scoring has proven to be useful for pain evaluation in humans. In the last decade, equivalent scales have been developed for various animal species, including large domestic animals. The research question of this systematic review was as follows: is facial expression scoring (intervention) a valid method to evaluate pain (the outcome) in large domestic animals (population)? Method: We searched two databases for relevant articles using the search string: "grimace scale" OR "facial expression" AND animal OR "farm animal" NOT "mouse" NOT "rat" NOT "laboratory animal." The risk of bias was estimated by adapting the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) checklist. Results: The search strategy extracted 30 articles, with the major share on equids and a considerable number on cows, pigs, and sheep. Most studies evaluated facial action units (FAUs), including the eye region, the orbital region, the cheek or the chewing muscles, the lips, the mouth, and the position of the ears. Interobserver reliability was tested in 21 studies. Overall FAU reliability was substantial, but there were differences for individual FAUs. The position of the ear had almost perfect interobserver reliability (interclass coefficient (ICC): 0.73-0.97). Validity was tested in five studies with the reported accuracy values ranging from 68.2 to 80.0%. Discussion: This systematic review revealed that facial expression scores provide an easy method for learning and reliable test results to identify whether an animal is in pain or distress. Many studies lack a reference standard and a true control group. Further research is warranted to evaluate the test accuracy of facial expression scoring as a live pen side test.

Bored at home?-A systematic review on the effect of environmental enrichment on the welfare of laboratory rats and mice.

Boredom is an emotional state that occurs when an individual has nothing to do, is not interested in the surrounding, and feels dreary and in a monotony. While this condition is usually defined for humans, it may very well describe the lives of many laboratory animals housed in small, barren cages. To make the cages less monotonous, environmental enrichment is often proposed. Although housing in a stimulating environment is still used predominantly as a luxury good and for treatment in preclinical research, enrichment is increasingly recognized to improve animal welfare. To gain insight into how stimulating environments influence the welfare of laboratory rodents, we conducted a systematic review of studies that analyzed the effect of enriched environment on behavioral parameters of animal well-being. Remarkably, a considerable number of these parameters can be associated with symptoms of boredom. Our findings show that a stimulating living environment is essential for the development of natural behavior and animal welfare of laboratory rats and mice alike, regardless of age and sex. Conversely, confinement and under-stimulation has potentially detrimental effects on the mental and physical health of laboratory rodents. We show that boredom in experimental animals is measurable and does not have to be accepted as inevitable.

Investigations on the Transfer of Quinolizidine Alkaloids from Lupinus angustifolius into the Milk of Dairy Cows.

Lupin varieties with a low content of quinolizidine alkaloids (QAs) like blue sweet lupin (BSL) have long been used as a protein source for dairy cows. A health concern for humans may arise from the transfer of acute toxic QAs from feed into cow's milk. This study is the first to quantify the transfer of QAs from BSL into cow's milk with experimental and modeling methods. Four lactating dairy cows were subjected to two 7 day feeding periods with 1 and 2 kg/d BSL, respectively, each followed by a depuration period. BSL contained 1774 mg/kg dry matter total QAs. Individual milk samples were taken twice daily and QA contents in feed and milk determined with liquid chromatography-tandem mass spectrometry. Transfer of QAs into the milk was already seen with the administration of 1 kg/d BSL, with differences in transfer rates (TRs) between individual QAs. A toxicokinetic model was derived to quantify and predict QA feed-to-food transfer. For the four most prominent QAs, our model shows an α-half-life of around 0.27 d. TRs were obtained for six QAs and were between 0.13 (sparteine) and 3.74% (multiflorine). A toxicological assessment of milk containing QAs as measured in this study indicated a potential health concern.

Using an OSCE to Explore the Role of Structured Debriefing and Self-Directed Learning in Simulator-Based Clinical Skill Training in Production Animal Reproductive Medicine.

Self-directed learning is associated with several benefits in simulation-based clinical skill training and can be complemented by feedback in the form of post-event debriefing. In this study, final-year veterinary medicine students (n = 111) were allocated into one of three groups and practiced four clinical skills from the domain of production animal reproductive medicine in a clinical skills laboratory. Group 1 completed an instructor-led practice session (I), group 2 completed a self-directed practice session with post-event debriefing (D), and group 3 completed a self-directed practice session without debriefing (control, C). Each practice session included two clinical skills categorized as being directly patient-related (patient) and two clinical skills involving laboratory diagnostics or assembling equipment (technical). Students evaluated the practice session using Likert-type scales. Two days after practice, 93 students took part in an objective structured clinical examination (OSCE). Student performance was analyzed for each learning station individually. The percentage of students who passed the OSCE did not differ significantly between the three groups at any learning station. While the examiner had an effect on absolute OSCE scores (%) at one learning station, the percentage of students who passed the OSCE did not differ between examiners. Patient learning stations were more popular with students than technical learning stations, and the percentage of students who passed the OSCE was significantly larger among students who enjoyed practicing at the respective station (90.9%) than among those who did not (77.8%).

Calving Management: A Questionnaire Survey of Veterinary Subject Matter Experts and Non-Experts.

Accurate detection of the onset of parturition is a key factor in the prevention of dystocia. In order to establish current best practice recommendations for calving management, we asked subject matter experts (SME) who had published on calving management (by online survey, n = 80) and non-SMEs, veterinary practitioners (by workshop survey, n = 24) for their opinions. For this, we designed a questionnaire on the significance of signs of imminent parturition (SIP), the frequency of calving observation, and influencing factors for the timing of cow movement to a maternity pen. The response rate was 67.5% in the online survey and 100% in the workshop survey. The majority (89.7%) of all respondents agreed that it is beneficial for successful calving management to differentiate between stage I and II of parturition. Of 12 signs of imminent parturition (for stage I and II), "restlessness" and "visibility of fetal parts in vulva" were cited by 56.5% and 73.3% of SME and non-SME respondents, respectively. There was no consensus on the right time to move the cow to the maternity pen; recommendations varied from one to over 21 days. Almost half of the respondents (45.7%) recommended a 6-h observation interval for prepartum cows in the maternity pen. This study identified a strong consensus on the SIP and how and when to observe cows prior to parturition. SMEs and non-SMEs provided broadly similar recommendations, while the SMEs and the non-SMEs differed significantly in the number of publications on calving they authored, they differed little in their knowledge of calving management.

Examining the Role of Structured Debriefing in Simulator-Based Clinical Skills Training for Namibian Veterinary Students: A Pilot Study.

Post-event debriefing has been described as an effective tool in improving learning achievements in simulator-based teaching. This article examines the effect of structured post-event debriefing sessions in simulator-based veterinary clinical skills training. Nineteen Namibian veterinary students took part in instructor-led practice, self-directed practice with structured post-event debriefing and self-directed practice without debriefing (control) at three different learning stations in a veterinary clinical skills laboratory. Students evaluated their practice experience using Likert-type scales, and learning achievements were assessed using an objective structured clinical examination (OSCE). The results show that the choice of practice model had no significant effect on learning achievements overall. However, at individual learning stations, different practice models showed significant differences regarding effect on learning achievements. Students generally preferred practice sessions with some form of instructor involvement but the importance of instructor guidance was rated differently at each individual learning station.

[Taking body temperature in cattle - critical evaluation of an established diagnostic test]. / Messung der Körpertemperatur beim Rind ­ eine altbewährte Methode kritisch hinterfragt.

Measuring the body temperature is an essential component of the clinical examination of bovines. Next to its value for the diagnosis of diseases, body temperature also is an important value in clinical studies concerning estrus detection, calving time point prediction, or the evaluation of heat stress. This systematic review critically evaluates different methods of measuring bovine body temperature including rectal measurement, vaginal or ruminal temperature loggers, milk temperature, and infrared body surface thermography. Although body temperature measurement is a commonly employed and established diagnostic test, a close look at scientific studies displays multiple factors influencing body temperature data. The variability of results emphasizes the relevance of critically evaluating new measuring methods before introducing them into research or routine practice. Especially in terms of identifying specific cut-off values, i. e. for fever defined as body temperature > 39.5°C, the precision of the method is of importance, as the acquired values possess a high degree of influence on the veterinary decision taking process.

Effect of denaverine hydrochloride application to heifers on the APGAR score and lactate concentration in newborn calves.

OBJECTIVE: The aim of this study was to evaluate the influence of Denaverine hydrochloride (DNH) in heifers on calf vitality. MATERIAL AND METHODS: A total of 83 calvings with 38 female and 45 male calves were included in the study. Thirty minutes after onset of stage 2 of calving, 400 mg DNH or placebo (0.9% NaCl) were administered subcutaneously. If the calving procedure was not completed after 60 minutes, an extraction was conducted and pulling force was measured by using a digital force gauge. Directly after parturition, vitality of calves was evaluated using a modified APGAR score. Additionally, lactate concentration in blood from Vena auricularis was measured with a handheld measuring device (lactate scout). RESULTS: No effect of treatment was observed on APGAR score and lactate concentration. CLINICAL RELEVANCE: Denaverine hydrochloride is a regularly used substance in obstetrics in veterinary medicine in many European countries. We could not confirm our hypotheses that treating heifers with DNH has a positive effect on calf vitality evaluated by APGAR score and lactate concentration in blood.

A Proof of Concept: Are Detection Dogs a Useful Tool to Verify Potential Biomarkers for Lung Cancer?

Early and reliable diagnostic test is essential for effective therapy of lung cancer. Volatile organic compounds that are characteristic for cancer could serve as valuable biomarkers in cancer diagnosis. Both trace analytical and detection dog approaches give some evidence for the existence of such biomarkers. In this proof of concept, study dogs and trace analysis were implemented in combination to gain more information concerning cancer biomarkers. Two dogs were trained to distinguish between absorbed breath samples of lung cancer patients and healthy persons and succeeded with correct identification of patients with 9/9 and 8/9 and correct negative indications from of 8/10 and 4/10 samples from healthy individuals. A recent observational study found that breath samples from lung cancer patients showed an increase in 1-butanol, 2-butanone, 2-pentanone, and hexanal. Synthetic air samples were therefore fortified with these compounds and adsorbed to a fleece. Tested against breath samples from healthy probands, on presentation to the dogs these synthetic samples provoked an indication in three out of four samples. We were able to demonstrate that a combination of the natural nose of a dog and a trace analytic technique can be a valuable concept in the search for cancer biomarkers.

Odor Perception by Dogs: Evaluating Two Training Approaches for Odor Learning of Sniffer Dogs.

In this study, a standardized experimental set-up with various combinations of herbs as odor sources was designed. Two training approaches for sniffer dogs were compared; first, training with a pure reference odor, and second, training with a variety of odor mixtures with the target odor as a common denominator. The ability of the dogs to identify the target odor in a new context was tested. Six different herbs (basil, St. John's wort, dandelion, marjoram, parsley, ribwort) were chosen to produce reference materials in various mixtures with (positive) and without (negative) chamomile as the target odor source. The dogs were trained to show 1 of 2 different behaviors, 1 for the positive, and 1 for the negative sample as a yes/no task. Tests were double blind with one sample presented at a time. In both training approaches, dogs were able to detect chamomile as the target odor in any presented mixture with an average sensitivity of 72% and a specificity of 84%. Dogs trained with odor mixture containing the target odor had more correct indications in the transfer task.

A survey of drying-off practices on commercial dairy farms in northern Germany and a comparison to science-based recommendations.

While dry cow management is important for health, milk production and fertility information on drying-off procedures implemented on commercial dairy farms is lacking. Current drying-off management procedures on commercial dairy farms were evaluated using a questionnaire and results compared with recommendations given in the current literature. Ninety-one participants from a farmer education event completed the survey. On average, cows were dried off seven weeks before calving. Only 9.9 per cent of the farms had a dry period length of five weeks or less. A continuous milking regime without dry period was not established on any farm participating in the survey. Most farmers performed an abrupt drying-off (73.0 per cent). Only 11.8 and 15.0 per cent attempted to lower milk yield prior to drying-off by reducing milking frequencies and adjusting feed rations, respectively. While a blanket antibiotic dry cow treatment was carried out on 79.6 per cent of the farms, selective dry cow treatment was not mentioned by any farmer. Although 77.4 per cent preponed the drying-off date in low-yielding cows, an altered drying-off procedure in high-yielding dairy cows was rare (9.7 per cent). This survey provides an insight into drying-off procedures currently applied on commercial dairy farms in northern Germany.

In vitro cultured lung cancer cells are not suitable for animal-based breath biomarker detection.

In vitro cultured lung cancer cell lines were investigated regarding the possible identification of volatile organic compounds as potential biomarkers. Gas samples from the headspace of pure culture medium and from the cultures of human lung adenocarcinoma cell lines A549 and Lu7466 were exposed to polypropylene fleece in order to absorb odour components. Sniffer dogs were trained with loaded fleeces of both cell lines, and honey bees were trained with fleeces exposed to A549. Afterwards, their ability to distinguish between cell-free culture medium odour and lung cancer cell odour was tested. Neither bees nor dogs were able to discriminate between odours from the cancer cell cultures and the pure culture medium. Solid phase micro extraction followed by gas chromatography with mass selective detection produced profiles of volatiles from the headspace offered to the animals. The profiles from the cell lines were largely similar; distinct differences were based on the decrease of volatile culture medium components due to the cells' metabolic activity. In summary, cultured lung cancer cell lines do not produce any biomarkers recognizable by animals or gas chromatographic analysis.