Federation of European Laboratory Animal Science Associations recommendations of best practices for the health management of ruminants and pigs used for scientific and educational purposes

Most ruminants and pigs used for scientific and educational aims are bred not for these purposes but in a farm environment. Given the wide range of diseases that these species might have, ensuring that the animals’ health status is appropriate can be complex and challenging. The Federation of European Laboratory Animal Science Associations has previously published recommendations for the health monitoring of experimental colonies of pigs (1998) and, respectively, calves, sheep and goats (2000). Unfortunately, the uptake of those recommendations was poor and insufficiently reported in scientific publications. These new recommendations for best practice focus on the main species of ruminants (cattle, sheep and goats) and pigs. They provide general and specific information helpful for designing a health management programme for the suppliers and for the user establishments, as well as guidance on animal procurement. Critical thinking based on the fields of use of the animals is promoted, aiming to help in taking informed decisions rather than establishing an exhaustive exclusion list for pathogens. Implementing the best health and welfare management practices should be done under the guidance of a competent attending veterinarian, with expertise and sufficient authority to take the appropriate action, doubled by excellent communication skills. It is strongly recommended that the user establishment’s veterinarian works in close collaboration with the supplier’s veterinarian. // Die meisten Wiederkäuer und Schweine, die zu wissenschaftlichen und Ausbildungszwecken dienen, werden nicht eigens dafür, sondern in einem landwirtschaftlichen Umfeld gezüchtet. Angesichts des breiten Spektrums potenzieller Krankheiten bei diesen Tierarten kann die Gewährleistung eines adäquaten Gesundheitszustands der Tiere durchaus komplex und schwierig sein. FELASA hat bereits früher Empfehlungen für die Gesundheitsüberwachung von Versuchskolonien von Schweinen (1998) bzw. Kälbern, Schafen und Ziegen (2000) veröffentlicht. Leider stießen diese Empfehlungen auf mangelndes Echo und wissenschaftliche Publikationen berichteten diesbezüglich nur unzureichend. Die vorliegenden neuen Empfehlungen für beste Praxis konzentrieren sich auf die wichtigsten Arten von Wiederkäuern (Rinder, Schafe und Ziegen) sowie auf Schweine. Sie enthalten allgemeine und spezifische Informationen, die für die Gestaltung eines Gesundheitsmanagementprogramms für die Lieferanten und für die Verwendereinrichtungen nützlich sind, ebenso wie Hinweise zur Tierbeschaffung. Kritisches Denken auf der Grundlage der Anwendungsbereiche der Tiere, das darauf abzielt, fundierte Entscheidungen zu treffen, anstatt eine erschöpfende Ausschlussliste für Krankheitserreger zu erstellen, wird unterstützt. Die Umsetzung der besten Gesundheits- und Tierschutzmanagementpraktiken sollte unter der Anleitung eines kompetenten behandelnden Tierarztes erfolgen, der über Fachwissen und genügend Autorität sowie über ausgezeichnete Kommunikationsfähigkeiten verfügen, um die entsprechenden Maßnahmen zu ergreifen und zu vermitteln. Eine enge Zusammenarbeit zwischen dem Tierarzt der Verwendereinrichtung und dem Tierarzt des Lieferanten wird dringend empfohlen

A novel closed-chest porcine model of chronic ischemic heart failure suitable for experimental research in cardiovascular disease

Cardiac pathologies are among the leading causes of mortality and morbidity in industrialized countries, with myocardial infarction (MI) representing one of the major conditions leading to heart failure (HF). Hitherto, the development of consistent, stable, and reproducible models of closed-chest MI in large animals, meeting the clinical realism of a patient with HF subsequent to chronic ischemic necrosis, has not been successful. We hereby report the design and ensuing application of a novel porcine experimental model of closed-chest chronic ischemia suitable for biomedical research, mimicking post-MI HF. We also emphasize the key procedural steps involved in replicating this unprecedented model, from femoral artery and vein catheterization to MI induction by permanent occlusion of the left anterior descending coronary artery through superselective deployment of platinum-nylon coils, as well as endomyocardial biopsy sampling for histologic analysis and cell harvesting. Our model could indeed represent a valuable contribution and tool for translational research, providing precious insights to understand and overcome the many hurdles concerning, and currently quenching, the preclinical steps mandatory for the clinical translation of new cardiovascular technologies for personalized HF treatments

A percutaneous method for single catheter recording of multiple Monophasic action potentials from the epicardial surface in intact anesthetized rodents

A surgical method for programmed electrical stimulation (PES) in mice has been reported by Gutstein et al (2003). We developed a percutaneous approach to record multiple epicardial monophasic action potentials (multi-MAP) in rodents, with a single amagnetic catheter (AC). Methods: Under 6 ECG leads monitoring the AC was introduced with a sub-xyphoid puncture, in 5 anesthetized, spontaneously breathing, Wistar rats (WR) and 5 guinea pigs (GP). The AC was moved at several epicardial sites. 4 MAPs were simultaneously recorded at each site with a fixed (1 mm) inter-electrode distance. MAP signals, differentially amplified and filtered at DC-500 Hz were digitized at 1kHz. The same AC was used for PES at pacing cycles between 200 and 250 ms. Ventricular effective refractory period (VERP) was evaluated with the accuracy of 2 ms. MAP duration (MAPd), at 50% and 90% levels of repolarization, and local activation time among the four MAPs were calculated. Results: All but one animal tolerated the procedure and survived. One rat died for respiratory arrest at the moment of the puncture. One rat was restudied after 6 months and survived also the second procedure. MAPd 90% (ms) were: 69\ub15.6 (WR) and 172\ub112.3 (GP). VERP were: 67\ub19.4 (WR) and 155\ub115.6 (GP). Conclusion: The percutaneous approach avoids cut-down, is well tolerated and keeps animals alive and available for longitudinal investigations. The multi-MAP method provides information about arrhythmogenic mechanisms (i.e. MAPd/ERP ratio, detection of ventricular repolarization (VR) inhomogeneity and of local block), with high spatial resolution. This animal model can be used for correlative studies between dispersion of MAP duration and surface estimate of QT dispersion and T wave abnormalities

Contactless magnetocardiographic study of age- and gender-related variability of ventricular repolarization parameters in guinea pigs

Guinea pigs (GPs) are commonly used for preclinical investigation of the ability of new drugs to induce QT alterations and arrhythmias, since their myocytes have an action potential with a well-defined plateau and most of the ion currents present in humans. The aim of this study was to evaluate the accuracy of multi-site magnetocardiography (MCG) for the assessment of age-related variation of ventricular repolarization maps in GPs. Methods: 20 adult GPs (10 males and 10 females) were investigated with the same unshielded 36-channel MCG instrumentation used for clinical recordings, at the age of 5 months [weight: 268.1 ± 19 gr]. Only 12 survived until the age of 14 months [weight: 595.6 ± 90.5 gr] and were restudied. RR, PR, QRS, QTpeak, QT end, JTpeak, JT end and Tpeak-end intervals were measured from MCG waveforms. MCG imaging was automatically obtained as MF maps, and with the Equivalent Current Dipole (ECD) and Effective Magnetic Dipole inverse solutions. Results: In the 12 animals, available for longitudinal MCG evaluation, a statistically significant age-related prolongation of the QRS (p < 0.01) and of the P-R (p < 0.05) intervals was found. On the contrary, neither significant age-related variations of ventricular repolarization (VR) intervals, nor gender-related differences, were found. MF maps calculated at the peaks of the P and R waves were similar in all animals, while the MF pattern at the peak of the T-wave widely varied, at 5 months. At 14 months, the MF distribution at the Tpeak remained unchanged in some animals, but markedly changed in others, with rotation of the dipolar T-wave pattern, in the absence of variations of P and QRS MF. The strength of the ECD, measured at the peak of the P, QRS and T waves, was stronger (p<0.01), at the age of 14 months than at the age of 5 months. Conclusions: MCG is reliable for VR assessment in GP. In contrast from findings in rats, age-related changes and gender-related differences of VR parameters were not statistically significant in GPs. Further works is deserved to understand the physiological meaning of VR MF variability observed in our healthy GP

Longitudinal study of cardiac electrical activity in anesthetized guinea pigs by contactless magnetocardiography

Guinea pigs (GPs) are used for preclinical evaluation of electrophysiologic effects of new drugs, because their myocytes have human-like action potentials and ventricular repolarization\u2019s (VR) ion currents. This study was aimed to assess the reliability of magnetocardiographic (MCG) mapping for longitudinal studies of GP cardiac electrical activity. Eighteen anesthetized GPs were investigated with an unshielded 36-channel MCG instrumentation, at the age of 5 months (268.1 \ub1 19 g). Twelve GPs survived and were restudied when 14 months old (595.6 \ub1 90.5 g). RR, PR, QRS, QTpeak, QTend, JTpeak, JTend and Tpeak-end intervals were measured from MCG waveforms. Magnetic field (MF) maps, equivalent current dipole (ECD) parameters and current density imaging were also analyzed. A significant prolongation of the PR (p < 0.05) and QRS (p < 0.001) intervals was found at 14 months. Gender-related differences of VR intervals were not significant. Ppeak and QRSpeak MFs were similar in all animals, while Tpeak MF varied interindividually at 5 months and showed a rotation in some animals, at 14 months. The ECD strengths, measured at the Ppeak, QRSpeak and Tpeak were stronger (p < 0.01) at the age of 14 months than at 5 months. In contrast to findings in Wistar rats, age-related and gender-related differences of MCG VR parameters were not significant in GPs. Further work is necessary to clarify the variability of VR MF observed in healthy GP

Intake of Boron, Cadmium, and Molybdenum enhances rat thyroid cell transformation

Abstract Background Epidemiologic data in volcanic areas suggest that environmental factors might be involved in the increase of thyroid cancer (TC) incidence. Recent reports indicate that several heavy metals and metalloids are increased in volcanic areas. This study aims to evaluate the combined effect of three of these elements Boron (B), Cadmium (Cd), and Molybdenum (Mo) - all increased in the volcanic area of Mt. Etna, in Italy - on thyroid tumorigenesis in the rat. Methods Female Wistar rats prone to develop thyroid tumors by low-iodine diet and methimazole treatment received ad libitum drinking water supplemented with B, Cd, and Mo at concentrations in the range found in the urine samples of residents of the volcanic area. At 5 and 10 months animals were euthanized, and their thyroid analysed. Statistical analysis was performed with a 2-way unpaired t-test. Results No toxic effect of the three elements on the growth of the animals was observed. A significant increase of histological features of transformation was observed in thyroid follicular cells of rats treated with B, Cd, and Mo compared with those of control group. These abnormalities were associated with decreased iodine content in the thyroid. Conclusions This study provides the evidence that slightly increased environmental concentrations of B, Cd, and Mo can accelerate the appearance of transformation marks in the thyroid gland of hypothyroid rats

High Saturated-Fat Diet Induces Apoptosis in Rat Enterocytes and Blunts GIP and Insulin-Secretive Response to Oral Glucose Load

Lipoapoptosis has been described in many organs and tissues, but never in enterocytes. We hypothesized that a high saturated-fat diet can induce duodenal enterocyte apoptosis and impair gastric inhibitory polypeptide (GIP) secretion. Forty male Wistar rats, approximately 4 months old, were randomized on standard laboratory or purified tripalmitin-based high-fat diet (59% calories). An oral-glucose tolerance test was performed after 30 and 90 days of diet to measure plasma glucose, insulin and GIP. Duodena were processed for histology and immunohistochemistry by transferase-mediated dUTP nick end-labeling (TUNEL) method. Apoptosis was confirmed by enzyme-linked immunosorbent assay. Glycemic response was significantly higher (P &lt; 0.01 vs controls) in rats after 90 days. Insulin curve was markedly increased at 30 days, while it was blunted at 90 days. GIP area under the curve was 425.6 +/- 67.6 ng ml(-1) at 30 days vs 150.2 +/- 33.4 ng ml(-1) in controls (P &lt; 0.001) and dropped to 53.8 +/- 25.8 ng ml(-1) at 90 days (P &lt; 0.0001). TUNEL-positive nuclei were 66.08+/-26.19 at 30 days 57 (34.58+/-17 in controls, P &lt; 0.05) and 216.99 +/- 129.42 nuclei per mm(3) at 90 days (38.75 +/- 18.36 in controls, P &lt; 0.0001). A high saturated-fat diet stimulates GIP secretion but with time induces apoptosis of duodenal villi epithelium, showing for the first time that enterocytes are also prone to lipoapoptosis. The reduction of circulating GIP levels might contribute to hypoinsulinemia and hyperglycemia