Doping in Racing Pigeons (Columba livia domestica): A Review and Actual Situation in Belgium, a Leading Country in This Field.

Pigeon racing is a sport in which trained homing pigeons (Columba livia domestica) are released between 60 and 1200 km from their loft and then have to return home as quickly as possible. The first race was held in 1818 in Belgium and since then, Belgium has led the world in pigeon breeding. Unfortunately, as in other sports, doping has become a major issue and doping controls have been implemented. This review provides information about pigeon racing, rules from the Royal Federation Colombophile of Belgium, and laws applicable in Belgium as doping control issues cannot be understood without including them as part of pigeon racing. The main pharmacological data concerning corticoids, non-steroidal anti-inflammatory drugs, anabolic steroids, pain relievers and narcotic analgesics, bronchodilators and ß-agonists, drugs acting on the central nervous system and other performance-enhancing drugs, in addition to methods relevant to doping in pigeons are presented. Moreover, the chosen matrix and analytical methods are described.

Effects of a Single Opioid Dose on Gastrointestinal Motility in Rabbits (Oryctolagus cuniculus): Comparisons among Morphine, Butorphanol, and Tramadol.

The aim of this study was to evaluate and compare the effects of single doses of butorphanol, morphine, and tramadol on gastrointestinal motility in rabbits (Oryctolagus cuniculus) using non-invasive imaging methods, such as radiographic barium follow through and ultrasonographic contraction counts. Time-lapse radiographic and ultrasound examinations were performed before and after a single intramuscular dose of 5 mg kg-1 butorphanol, 10 mg kg-1 morphine, or 10 mg kg-1 tramadol. Pyloric and duodenal contraction counts by ultrasonography and radiographic repletion scores for the stomach and caecum were analysed using a mixed linear model. No significant effect was noted on ultrasound examinations of pyloric and duodenal contractions after administration of an opioid treatment. Morphine had a significant effect on the stomach and the caecum repletion scores, whereas butorphanol had a significant effect only on the caecum repletion score. Tramadol had no significant effect on the stomach or caecum repletion scores. The present findings suggest that a single dose of 5 mg kg-1 butorphanol or 10 mg kg-1 morphine temporarily slows gastrointestinal transit in healthy rabbits, preventing physiological progression of the alimentary bolus without the induction of ileus. In contrast, a single dose of 10 mg kg-1 tramadol has no such effects.

In vitro and in vivo assessment of eprociclovir as antiviral treatment against testudinid herpesvirus 3 in Hermann's tortoise (Testudo hermanni).

Tortoises belonging to the Testudinidae family are infected by Testudinid herpesviruses. Testudinid herpesvirus 3 (TeHV-3) is considered the most pathogenic and affects several tortoise species, particularly those from the Testudo genus. As most species of this genus are endangered contribute to ecological concerns over this virus. Here, we aimed to explore the rational development of an antiviral treatment against TeHV-3 using Hermann's tortoise (Testudo hermanni) as a host model. Ten antiviral compounds were tested in cell culture for their toxicity and their activity against TeHV-3. Eight compounds exhibited different levels of activity against TeHV-3 with either no or only minor cytotoxic effects on cells. Next, eprociclovir (EPV, ciprovir) was selected for further investigations in vivo. Its pharmacokinetic properties were investigated after a single sub-cutaneous administration at 5 or 10 mg/kg. Plasma concentrations remained above half maximal effective concentration (EC50) for 2.2 and 4.4 h after administration at 5 and 10 mg/kg, respectively. Finally, EPV toxicity was investigated after administration at the dose of 10 mg/kg, BID for seven consecutive days. As early as one day after initiation of the treatment up to its end, EPV plasma concentration remained under the EC50. Apathy and anorexia developed after 7 days. Biochemical and anatomopathological examinations revealed nephrotoxic effects of EPV. Altogether, these data suggest that EPV is not a suitable molecule for the treatment of TeHV-3. Further studies are required to determine whether the other molecules identified here for their anti-TeHV-3 activity represent potential candidates for the development of efficacious treatments.

Influence of a single dose of buprenorphine on rabbit (Oryctolagus cuniculus) gastrointestinal motility.

OBJECTIVE: To establish a noninvasive imaging protocol for rabbit gastrointestinal transit evaluation. To assess the effect of a single injection of buprenorphine on the digestive transit of rabbits via this new technique. STUDY DESIGN: Prospective, parallel study. ANIMALS: Fifteen specific pathogen-free male New Zealand White rabbits weighing 2.68 ± 0.28 kg. METHODS: A 10 mL kg-1 barium meal was administered and the rabbits were subjected to serial radiographic and ultrasound examinations without treatment and 1 week later following a single intramuscular dose of 100 µg kg-1 of buprenorphine. Radiographic data from the stomach and caecum were collected and assigned a retention score ranging from 0 (no barium) to 3 (large amount of barium). The resulting scores and pyloric and duodenal contraction counts were analysed using a mixed linear model and are expressed as least square mean (lsm) ± standard error. Transit was estimated based on the apparition time of faeces in the pelvic area and analysed using a Wilcoxon test. A p < 0.05 was considered significant. RESULTS: Buprenorphine treatment induced a higher lsm number of pyloric (1.73 ± 0.19 versus 0.78 ± 0.19, p < 0.01) and lsm duodenal contractions (17.35 ± 1.04 versus 13.44 ± 1.04, p < 0.01). Buprenorphine administration decreased the lsm barium retention score in the stomach (2.44 ± 0.05 versus 2.64 ± 0.05, p < 0.01), but had no effect on the lsm barium retention score in the caecum. The time to apparition of faeces in the pelvic area was not influenced by buprenorphine administration (p = 0.66). CONCLUSIONS AND CLINICAL RELEVANCE: A single high dose of buprenorphine appears to have no adverse effect on gastrointestinal motility in healthy rabbits.

The Genome of a Tortoise Herpesvirus (Testudinid Herpesvirus 3) Has a Novel Structure and Contains a Large Region That Is Not Required for Replication In Vitro or Virulence In Vivo.

UNLABELLED: Testudinid herpesvirus 3 (TeHV-3) is the causative agent of a lethal disease affecting several tortoise species. The threat that this virus poses to endangered animals is focusing efforts on characterizing its properties, in order to enable the development of prophylactic methods. We have sequenced the genomes of the two most studied TeHV-3 strains (1976 and 4295). TeHV-3 strain 1976 has a novel genome structure and is most closely related to a turtle herpesvirus, thus supporting its classification into genus Scutavirus, subfamily Alphaherpesvirinae, family Herpesviridae. The sequence of strain 1976 also revealed viral counterparts of cellular interleukin-10 and semaphorin, which have not been described previously in members of subfamily Alphaherpesvirinae. TeHV-3 strain 4295 is a mixture of three forms (m1, m2, and M), in which, in comparison to strain 1976, the genomes exhibit large, partially overlapping deletions of 12.5 to 22.4 kb. Viral subclones representing these forms were isolated by limiting dilution assays, and each replicated in cell culture comparably to strain 1976. With the goal of testing the potential of the three forms as attenuated vaccine candidates, strain 4295 was inoculated intranasally into Hermann's tortoises (Testudo hermanni). All inoculated subjects died, and PCR analyses demonstrated the ability of the m2 and M forms to spread and invade the brain. In contrast, the m1 form was detected in none of the organs tested, suggesting its potential as the basis of an attenuated vaccine candidate. Our findings represent a major step toward characterizing TeHV-3 and developing prophylactic methods against it. IMPORTANCE: Testudinid herpesvirus 3 (TeHV-3) causes a lethal disease in tortoises, several species of which are endangered. We have characterized the viral genome and used this information to take steps toward developing an attenuated vaccine. We have sequenced the genomes of two strains (1976 and 4295), compared their growth in vitro, and investigated the pathogenesis of strain 4295, which consists of three deletion mutants. The major findings are that (i) TeHV-3 has a novel genome structure, (ii) its closest relative is a turtle herpesvirus, (iii) it contains interleukin-10 and semaphorin genes (the first time these have been reported in an alphaherpesvirus), (iv) a sizeable region of the genome is not required for viral replication in vitro or virulence in vivo, and (v) one of the components of strain 4295, which has a deletion of 22.4 kb, exhibits properties indicating that it may serve as the starting point for an attenuated vaccine.

Improving adjuvant systems for polyclonal egg yolk antibody (IgY) production in laying hens in terms of productivity and animal welfare.

The antibody production in the egg yolks of immunized laying hens is seen as a way of improving animal welfare compared with conventional production by mammals. Immunoglobulin Y (IgY) technology, however, has still to address welfare issues linked to the widespread use of an adjuvant in vaccines. Currently, Freund's adjuvants, complete (FCA) or incomplete (FIA), remain the standard. This study sought to evaluate various approaches used to enhance egg yolk antibody production in terms of both productivity and avian welfare. The outer membrane protein (OMP) of Salmonella Typhimurium was used as the prototype antigen. At 20 weeks of age, 56 ISA Brown hens, with specific-Salmonella-free status, were divided into seven groups (n=8) and received an initial intramuscular immunization. Hens in the two negative control groups received phosphate buffered saline (PBS) or FIA alone. Hens in the other groups received 80µg of Salmonella OMP emulsified with one of the following adjuvants: 200µl of FIA alone (T1); 200µl of FIA supplemented with 8µg of C-phosphate-guanosine oligodeoxynucleotides (CpG-ODN) (T2); and 280µl of Montanide ISA 70 VG (T4). Birds in the T3 group received the antigen in emulsion with FIA and were given the tested immunostimulatory component (l-carnitine) via their feed (100mg/kg). A positive control group (PC) received FCA for the first and final immunizations and FIA for the other boosters. Immunization was repeated after 20, 46, 82 and 221 days. Eggs were collected regularly until 242 days after the first immunization and the anti-Salmonella Typhimurium activities in the yolk were determined by ELISA. After 242 days, the birds were euthanized and the injection sites were evaluated for gross and microscopic lesions. Among the tested immunostimulatory approaches, supplementation of FIA with CpG-ODN led to a significant and long-lasting enhancement of the specific antibody response. This treatment was even higher than the positive benchmark using FCA in the first immunization. The study results showed that a clinical examination of injection sites is insufficient for drawing conclusions about the local tolerance of vaccines. Tissue damage was noticeable in all treatment groups. The birds receiving the Montanide adjuvant, however, had fewer and less severe lesions. Given these limited side-effects, Montanide ISA 70 VG could provide the depot effect needed to ensure the immunomodulatory efficiency of CpG-ODN. The association of these two adjuvants could prove a promising alternative to Freund's adjuvants (FA).

Detection of Usutu virus in a bullfinch (Pyrrhula pyrrhula) and a great spotted woodpecker (Dendrocopos major) in north-west Europe.

In October 2012, a 3-year-old bullfinch (Pyrrhula pyrrhula) held in captivity for its entire lifespan and a wild adult great spotted woodpecker (Dendrocopos major), both with neurological signs, were found 4 km from each other and 5 days apart in the Meuse Valley, Belgium. Non-suppurative encephalitis and mild degeneration and necrosis were identified in the brain and cerebellum, and Usutu virus antigen and RNA were detected by immunohistochemistry and real-time reverse transcriptase PCR, respectively. The two cases reported here represent the most western distribution of clinical disease in birds due to Usutu virus in Europe.

Epizootic rabbit enteropathy inoculum (TEC4): antibiograms and antibiotic fractionation.

Epizootic rabbit enteropathy (ERE) emerged and spread in Europe within the last 13 years causing major economical loss. The aims of the study was to evaluate antibiograms of TEC4, an inoculum composed of an extract of intestinal content of affected rabbits, and to test the potential of different antibiotic-based TEC4 fractions to reproduce the disease. Twenty nine different antibiotic discs were incubated for determining bacteria resistance. In a complementary study, nine tubes of liquid medium were inoculated with TEC4, incubated and added individually with amoxicillin/clavulanic acid, bacitracin, ceftiofur, doxycycline, novobiocin, streptomycyin, tylosin, vancomycin and 0.9% saline solution as control. The content of each tube was washed by centrifugation and suspended in saline. The three most effective antibiotics are florfenicol, amoxycillin/clavulanic acid and tylosin. A high concentration of Clostridium sordelli and Bacillus firmus were isolated in all fractions. Species never cultured from TEC4 were identified as Fusobacterium necrogenes (in vancomycin fraction), Cellulomonas sp (in novobiocin fraction) and Bacteroides distasonis (in doxycycline fraction). The ERE was reproduced when bacitracin, doxycycline and 0.9% fractions were inoculated. Rabbits showed ERE clinical signs with the specific drop in daily weight gain.