Accumulation of the pathogenic fungus Batrachochytrium dendrobatidis on the regressing tail of midwife toads Alytes obstetricans undergoing metamorphosis

Batrachochytrium dendrobatidis (Bd) is a fungus infecting the skin of amphibians. On metamorphosing animals the infection is difficult to detect because of the limited information concerning the location of Bd on the animals during this stage. Histological investigation revealed that Bd accumulated on the reabsorbing tail of metamorphosing animals. This observation may facilitate the detection of Bd in metamorphosing amphibians.

Alfaxalone anesthesia by immersion in oriental fire-bellied toads (Bombina orientalis).

OBJECTIVES To establish an effective alfaxalone concentration to be used for bath immersion of fire-bellied toads (Bombina orientalis) and to describe its effects. STUDY DESIGN Prospective experimental study. ANIMALS Thirteen oriental fire-bellied toads. METHODS The study was carried out in two phases. The pilot phase involved five animals and aimed to identify an alfaxalone concentration capable of producing induction of anesthesia, defined as immobility with a head down position and loss of responsiveness to stimulation with a stick. The following trial in an additional eight toads used the effective alfaxalone concentration established during the pilot phase. Data from 11 animals (three toads in the pilot study and the eight additional toads) were analyzed. Twenty minutes after immersion in the anesthetic solution, the toads were removed from the bath, and heart rate, respiratory rate, the righting, myotactic and the nociceptive withdrawal reflexes were evaluated every 5 minutes. The loss of both righting and nociceptive withdrawal reflexes was considered indicative of a surgical depth of anesthesia. The time elapsed from anesthetic induction to return of righting reflex, the quality of recovery and the occurrence of undesired effects were observed and recorded. RESULTS Immersion was found to be a suitable anesthetic technique for oriental fire-bellied toads and 200 mg L(-1) alfaxalone concentration produced anesthetic induction in 10 out of 11 toads. Side effects, such as skin irritation, erythema and changes in cutaneous pigmentation, were not observed in any animal. The duration of anesthesia ranged from 10 to 30 minutes after removal of the toads from the alfaxalone bath, and surgical depth of anesthesia was never achieved. CONCLUSIONS AND CLINICAL RELEVANCE It was concluded that alfaxalone anesthesia induced by immersion in a concentration of 200 mg L(-1) is only suitable for toads undergoing non-invasive short procedures.

Alfaxalone versus alfaxalone-dexmedetomidine anaesthesia by immersion in oriental fire-bellied toads (Bombina orientalis)

OBJECTIVE To determine a dexmedetomidine concentration, to be added to an alfaxalone-based bath solution, that will enhance the anaesthetic and analgesic effects of alfaxalone; and to compare the quality of anaesthesia and analgesia provided by immersion with either alfaxalone alone or alfaxalone with dexmedetomidine in oriental fire-bellied toads (Bombina orientalis). STUDY DESIGN Pilot study followed by a prospective, randomized, experimental trial. ANIMALS Fourteen oriental fire-bellied toads. METHODS The pilot study aimed to identify a useful dexmedetomidine concentration to be added to an anaesthetic bath containing 20 mg 100 mL(-1) alfaxalone. Thereafter, the toads were assigned to one of two groups, each comprising eight animals, to be administered either alfaxalone (group A) or alfaxalone-dexmedetomidine (group AD). After immersion for 20 minutes, the toads were removed from the anaesthetic bath and the righting, myotactic and nociceptive reflexes, cardiopulmonary variables and von Frey filaments threshold were measured at 5 minute intervals and compared statistically between groups. Side effects and complications were noted and recorded. RESULTS In the pilot study, a dexmedetomidine concentration of 0.3 mg 100 mL(-1) added to the alfaxalone-based solution resulted in surgical anaesthesia. The toads in group AD showed higher von Frey thresholds and lower nociceptive withdrawal reflex scores than those in group A. However, in group AD, surgical anaesthesia was observed in two out of eight toads only, and induction of anaesthesia was achieved in only 50% of the animals, as compared with 100% of the toads in group A. CONCLUSIONS AND CLINICAL RELEVANCE The addition of dexmedetomidine to an alfaxalone-based solution for immersion anaesthesia provided some analgesia in oriental fire-bellied toads, but failed to potentiate the level of unconsciousness and appeared to lighten the depth of anaesthesia. This limitation renders the combination unsuitable for anaesthetizing oriental fire-bellied toads for invasive procedures.

Alfaxalone-butorphanol versus alfaxalone-morphine combination for immersion anaesthesia in oriental fire-bellied toads (Bombina orientalis)

Oriental fire-bellied toads (Bombina orientalis) are small semi-aquatic anuran species popular as both pets and laboratory animals. Although they are commonly anaesthetized to undergo clinical and experimental procedures, very little is known about their anaesthetic management. The aims of this prospective, randomized, cross-over experimental trial were to establish effective butorphanol and morphine concentrations to be added to alfaxalone for immersion anaesthesia (pilot study), and to compare the anaesthetic and antinociceptive effects of the two drug mixtures (alfaxalone-butorphanol and alfaxalone-morphine), in Bombina orientalis toads. For the actual trial, the toads were randomly assigned to one of two treatment groups: AB and AM, with seven animals in each group, which received alfaxalone-butorphanol and alfaxalone-morphine combinations, respectively, at the concentrations established during the pilot study. Heart rate, respiratory rate, von Frey filament threshold and response to nociceptive withdrawal (NWR), righting and myotactic reflexes were measured at 5 min intervals until return of righting reflex was observed. The investigator who carried out all the measurements was blinded to the treatment. Any undesired effect or complication was noted and recorded. The two treatments were found to be comparable in terms of onset and duration of anaesthesia, and occurrence of undesired effects. However, group AM resulted in lower NWR scores and higher von Frey filament thresholds than group AB. It is concluded that, at the investigated concentrations and in combination with alfaxalone by immersion, morphine provides better antinociception than butorphanol in oriental fire-bellied toads.

Novel Features of the Prenatal Horn Bud Development in Cattle (Bos taurus).

Whereas the genetic background of horn growth in cattle has been studied extensively, little is known about the morphological changes in the developing fetal horn bud. In this study we histologically analyzed the development of horn buds of bovine fetuses between ~70 and ~268 days of pregnancy and compared them with biopsies taken from the frontal skin of the same fetuses. In addition we compared the samples from the wild type (horned) fetuses with samples taken from the horn bud region of age-matched genetically hornless (polled) fetuses. In summary, the horn bud with multiple layers of vacuolated keratinocytes is histologically visible early in fetal life already at around day 70 of gestation and can be easily differentiated from the much thinner epidermis of the frontal skin. However, at the gestation day (gd) 212 the epidermis above the horn bud shows a similar morphology to the epidermis of the frontal skin and the outstanding layers of vacuolated keratinocytes have disappeared. Immature hair follicles are seen in the frontal skin at gd 115 whereas hair follicles below the horn bud are not present until gd 155. Interestingly, thick nerve bundles appear in the dermis below the horn bud at gd 115. These nerve fibers grow in size over time and are prominent shortly before birth. Prominent nerve bundles are not present in the frontal skin of wild type or in polled fetuses at any time, indicating that the horn bud is a very sensitive area. The samples from the horn bud region from polled fetuses are histologically equivalent to samples taken from the frontal skin in horned species. This is the first study that presents unique histological data on bovine prenatal horn bud differentiation at different developmental stages which creates knowledge for a better understanding of recent molecular findings.

A 1.8-kb insertion in the 3'-UTR of RXFP2 is associated with polledness in sheep

Sheep breeds show a broad spectrum of different horn phenotypes. In most modern production breeds, sheep are polled (absence of horns), whereas horns occur mainly in indigenous breeds. Previous studies mapped the responsible locus to the region of the RXFP2 gene on ovine chromosome 10. A 4-kb region of the 3'-end of RXFP2 was amplified in horned and polled animals from seven Swiss sheep breeds. Sequence analysis identified a 1833-bp genomic insertion located in the 3'-UTR region of RXFP2 present in polled animals only. An efficient PCR-based genotyping method to determine the polled genotype of individual sheep is presented. Comparative sequence analyses revealed evidence that the polled-associated insertion adds a potential antisense RNA sequence of EEF1A1 to the 3'-end of RXFP2 transcripts.