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Animal experimentation is required to investigate complex physiological relationships and facilitates development of evidence-based knowledge. However, experimental protocols can interfere with the daily routine of the animals, result in stress and pain, and have adverse effects on health and production. The goal of this study was to investigate the effects of an intensive experimental protocol on health traits and production in transition dairy cows. Eighty experimental dairy Holstein cows (EXP group) underwent serial protocol-based clinical and ultrasonographic examinations, puncture of the jugular vein for blood collection or drug application, and liver biopsy samples, 14 d before until 42 d after parturition. Controls (CTR group) included 206 cows from the same herd, which fulfilled the same inclusion criteria and were kept under the same production management but were not handled for the purpose of this study. Procedure-related effects with a potentially negative effect on health and production were recorded. Furthermore, production, fertility and culling traits of the 2 groups (CTR, EXP) were compared. Most procedure-related adverse effects were associated with transcutaneous liver biopsies and included diffuse inflammation of the skin incision in 11.9% (42 of 320), abscessation of the skin or subcutis in 4.6% (11 of 240), and increased liver echogenicity of the biopsy site in 10.4% (27 of 240). The experimental procedures had a negative effect on milk yield at the start [days in milk (DIM) 5-50, difference: 2.3 kg, standard error (SE): 0.8 kg] and end of lactation (DIM 251-300, difference: 2.0 kg, SE: 1.0 kg; DIM 301-350, difference: 2.3 kg, SE: 1.2 kg) resulting in a lower 305-d milk yield in the EXP group than in the CTR group (difference: 472 kg, SE 214 kg). The milk fat % was higher in the EXP group than in the CTR group from 251 DIM onward (DIM 251-300, difference: 0.20%, SE: 0.09%; DIM 301-350, difference: 0.41%, SE: 0.17%). Also, the somatic cell score was higher in the EXP group than in the CTR group, during early (5-50 DIM, difference: 0.43, SE: 0.22) and from mid-lactation onward (DIM 151-200, difference: 0.43, SE: 0.2; DIM 201-250, difference: 0.49, SE: 0.22; DIM 251-300, difference: 0.55, SE: 0.25; DIM 301-350, difference: 0.61, SE: 0.28). Experimental procedures had no effect on first service conception rate and time to pregnancy, but had a positive effect on stillbirth rate with fewer stillbirths in the CTR group (0%) than in the EXP group (3.9%). Furthermore, experimental handling had no effect on time to culling or type of culling, whereby poor production was a more frequent reason noted for culling in the EXP group. Procedure-associated impairment of production in dairy cows is rarely reported and allows the estimation of the effects of such a study protocol on animal health and production. As a limitation for the interpretation of the results, the number of animals included and conduction in one single herd have to be considered.

IntroductionThe effect of a targeted pain management protocol consisting of sedation and local anesthesia on the stress response to treatment of dermatitis interdigitalis contagiosa (DINCO) was assessed in sheep placed in dorsal recumbency.MethodsBlood cortisol concentrations were measured once a day (Day -3 to 2) and additionally on day 0, six times during the claw treatment (stress model). Twelve healthy sheep (control group; HEALTHY) and 36 sheep with DINCO, randomly allocated to one of three treatment groups, underwent the stress model with or without pain control: the XYLA-IVRA sheep were sedated with 2% xylazine hydrochloride (XYLA) and received retrograde intravenous regional anesthesia (IVRA); the IVRA sheep underwent IVRA and received a placebo instead of sedation; the PLACEBO sheep received placebos for sedation and IVRA. The HEALTHY sheep underwent sham claw treatment and received placebos.Results and discussionThe cortisol concentrations were higher in sheep restrained in dorsal recumbency compared with the cortisol concentrations measured four hours later in standing sheep (HEALTHY 37.2 ± 3.3 ng/ml vs. 18.5 ± 3.3 ng/ml; DISEASED 34.0 ± 1.9 ng/ml vs. 17.6 ± 1.9 ng/ml; p < 0.001). The stress response of XYLA-IVRA (area under the curve; AUC = 34.9 ± 2.6 ng/ml) was reduced compared with the stress response of PLACEBO (AUC = 48.0 ± 2.6 ng/ml, p < 0.01) and HEALTHY sheep (AUC = 46.6 ± 2.5 ng/ml; p = 0.01). While cortisol concentration of XYLA-IVRA and HEALTHY sheep did not increase one day after the stress model (Day 1) compared with the day of the stress model (Day 0), both PLACEBO (47.4 ± 3.3 vs. 35.6 ± 3.1 ng/ml, p = 0.02) and IVRA sheep (39.1 ± 2.8 vs. 28.6 ± 3.1 ng/ml, p = 0.01) had higher cortisol concentrations. The results confirm that fixation in dorsal recumbency in a tilt squeeze chute was a major stressor in sheep. The differences in the cortisol concentration of the PLACEBO sheep versus the XYLA-IVRA sheep during and after the stress model illustrate the effect and necessity of pain management protocols in practice.

Clinical ketosis is common during the dairy cows' transition period and is responsible for considerable economic loss. Early identification of cows with subclinical ketosis is the first step for maintaining the health and productivity of dairy cows. The goals of the study were two-fold: The first was to examine the usefulness of a mobile test device as a cow-side test; and the second was to compare BHB concentrations measured by the ketometer using capillary blood and blood collected from the coccygeal vessels with values determined by a reference method in the laboratory using jugular blood.Blood samples were collected from a jugular vein or the coccygeal vessels in 81 dairy cows at 7 time points (14 and 7 days pre-partum and 7, 14, 21, 28, and 42 days post-partum) for kinetic enzymatic measurement of BHB concentration in the laboratory. Blood samples were concurrently collected from the coccygeal vessels or by pricking the vulvar lip at the transition of the skin to the mucosa (capillary blood) to determine BHB concentration using the WellionVet BELUA ketometer (MED TRUST GmbH, Marz, Austria).Initial errors in operating the ketometer were quickly eliminated with experience. BHB concentrations of jugular blood measured in the laboratory were 0.07 mmol/l lower than those measured in coccygeal blood. The mean BHB concentration measured in coccygeal and capillary blood using the WellionVet BELUA ketometer did not significantly differ but were 0.13 and 0.12 mmol/l respectively, lower than the mean jugular vein concentrations measured in the laboratory.The WellionVet BELUA ketometer is useful for determination of BHB concentration in cows provided that the manufacturer's specifications are followed. Capillary blood is best collected at the transition from the vulvar skin to its mucosa. The device generates rapid results that correlate well with BHB concentrations determined in the laboratory and with the results obtained from different blood collection sites. It is ideally suited for monitoring dairy cows for subclinical ketosis using capillary or coccygeal blood.ZIEL: Das vermehrte Auftreten von Ketose in der Transitphase bei Milchkühen verursacht hohe ökonomische Verluste. Die frühzeitige Identifizierung betroffener Tiere bereits im subklinischen Stadium ermöglicht, die Erkrankungshäufigkeit zu reduzieren und Verluste zu vermeiden. Ziel der Studie war, die Eignung eines mobilen Testgeräts zur Analyse der Betahydroxybutyrat-Konzentration (BHB-Konzentration) in der praktischen Anwendung zu prüfen. Darüber hinaus sollten die mit diesem Gerät gemessenen BHB-Konzentrationen mit denen verglichen werden, die mit einer Laborreferenzmethode aus dem Blut der V. jugularis ermittelt wurden.Bei 81 Milchkühen wurde zu 7 Zeitpunkten (14 und 7 Tage a. p. sowie 7, 14, 21, 28, 42 Tage p. p.) Blut für die Analyse der BHB-Konzentration entnommen. Zur Messung im Labor mit einer kinetisch-enzymatischen Methode diente Blut aus der V. jugularis und V. oder A. coccygea mediana, zur Messung mit dem Gerät WellionVet BELUA Blut aus der V. oder A. coccygea mediana sowie Kapillarblut (gewonnen am Übergang der Haut zur Schleimhaut an der Vulva).Anfängliche Anwendungsfehler bei der Bedienung des Geräts konnten mit Training abgestellt werden. Bei den Analyseergebnissen im Labor lagen die BHB-Konzentrationen aus dem Blut der V. jugularis um 0,07 mmol/l niedriger als die in Blut aus der V. oder A. coccygea mediana. Die mit dem Testgerät ermittelten BHB-Konzentrationen in Proben der V. oder A. coccygea mediana lagen um 0,13 mmol/l niedriger und die im Kapillarblut um 0,12 mmol/l niedriger als die im Labor aus dem Blut der V. jugularis gemessenen. Die mit dem Testgerät gemessenen BHB-Konzentrationen differierten zwischen den beiden Entnahmelokalisationen nicht signifikant.Für die Kapillarblutentnahme eignet sich die Lokalisation am Übergang von Haut zu Schleimhaut an der Vulva. Das Gerät WellionVet BELUA liefert sofort Ergebnisse mit guter Übereinstimmung zwischen den Messwerten in Proben von verschiedenen Blutentnahmeorten wie auch im Vergleich zur Laboranalyse. Das Testsystem ist zur Anwendung in Milchviehbetrieben geeignet. Als Matrix kann sowohl Kapillarblut als auch Blut aus der V. oder A. coccygea mediana empfohlen werden.

We studied the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in dairy calves as part of a routine health check protocol. In addition, data regarding antimicrobial use (AMU), farm hygiene, and farm management were collected in order to identify possible risks for ESBL occurrence. Ten farms participated in the study with a median of 781 milking cows (319-1701). All calves investigated were younger than two weeks with an average age of 6.8 (±3.9) days. The farms were visited and samples were collected twice at an interval of 7-11 months. Faecal samples diluted 1:10, were plated onto BrillianceTM ESBL agar in duplicates. After 24 hours at 37°C, colonies were counted and total colony forming units (cfu)/ml calculated. Bacteria species were identified biochemically. ESBL-production was phenotypically confirmed using the MICRONAUT-S β-Lactamases system. Additionally, antimicrobial susceptibility was tested using VITEK® 2 technology. Phylotyping of E. coli isolates and screening for bla genes was performed by PCR. ESBL-producing enterobacteria were detected on all farms and 96.5% of calves investigated shed ESBL-positive bacteria. Of all ESBL-producing isolates, the majority were E. coli (92.9%), followed by Enterobacter cloacae (5.1%) and Klebsiella pneumoniae subsp. pneumoniae (2.0%). The majority of E. coli isolates was clearly assigned to phylogroup C (25.0%), followed by phylogroups A (15.2%) and E (14.1%). CTX-M group 1 was most frequently detected (80.4%). E. cloacae contained blaCTX-M and blaTEM or blaSHV. K. pneumoniae harboured blaSHV only. Besides resistance to penicillins and cephalosporins, the majority of isolates was also resistant to one or more antibiotic classes, with a high proportion being resistant against fluoroqinolones. 52.5% of isolates were further characterised as threefold multidrug resistant gram-negative bacteria (3MDR-GNB) according to the German Commission for Hospital Hygiene and Infection Prevention. None of the isolates were 4MDR-GNB, i.e. none revealed carbapenem-resistance. Penicillins were the most frequently administered antibiotics to calves on most farms and were the predominant substance class at herd level on all farms. Overall, the number of calves treated prior to sampling was rather low (11.7%). Analyses of data regarding the farm management identified weaknesses in biosecurity and cleaning and disinfection. Besides beta-lactam antibiotics being the most commonly used antibiotics no other risk factors could be identified. In summary, the prevalence of ESBL-carriers in dairy calves was exceptionally high and should be motivation to develop strategies for the reduction of multidrug-resistant bacteria in farm animals.