A comparison of two intramuscular doses of xylazine–ketamine combination and tolazoline reversal in llamas

The anesthetic and cardiorespiratory effects of a low dose (LD, 0.4 mg kg^?1 xylazine and 4 mg kg^?1 ketamine) and a high dose (HD, 0.8 mg kg^?1 xylazine and 8 mg kg^?1 ketamine) of IM xylazine–ketamine combination were compared in a randomized cross‐over study using six castrated male llamas. Three llamas in each dosage group (LDT, HDT) were assigned to receive IM tolazoline (2 mg kg^?1) after 30 minutes of recumbency. All IM injections were given in the semitendinosus or semimembranosus muscles. Pulse, respiratory rate, and indirect arterial blood pressure were recorded every 10 minutes, and hemoglobin oxygen saturation was recorded every 5 minutes during lateral recumbency. Samples for arterial blood gas analysis were collected 5 minutes following recumbency and every 30 minutes thereafter. Base‐to‐apex ECG was monitored continuously. Analgesia was evaluated every 5 minutes by both a 30 minutes skin pinch and a needle prick of the toe. Most llamas breathed room air throughout anesthesia. Two llamas that developed severe hypoxemia (SpO₂ < 75%) received 5 minutes of nasal oxygen insufflation, but were maintained on room air for the rest of the anesthetic period. anova for repeated measures and Tukey's test were used to analyze cardiorespiratory data. Fischer's exact test was used to compare the ability of each to provide >30 minutes of lateral recumbency and analgesia. A p‐value < 0.05 was considered significant. Both dosages provided reasonably rapid induction following injection (LD: 10.8 ± 6.3 minutes; HD: 5.0 ± 1.1 minutes; p = 0.07). Duration of lateral recumbency and analgesia were 34.7 ± 6.7 and 27.3 ± 4.6 minutes, respectively, in the LDT llamas. None of the three remaining LD llamas remained in lateral recumbency for longer than 12 minutes. Duration of lateral recumbency and analgesia were 87.3 ± 18.5 and 67.7 ± 16.0 minutes, respectively, for the HD llamas that did not receive tolazoline. The HDT llamas were recumbent for a significantly shorter time (43.3 ± 0.6 minutes; p = 0.05). The ability to provide >30 minutes of recumbency and analgesia was better in the HD group (6/6) than in the LD group (2/6) (p = 0.03). No differences between dosages were seen in pulse rate, respiratory rate, or arterial pressures. No ECG abnormalities were seen. Transient hypoxemia was seen in the first 10 minutes of lateral recumbency in the HD group by both hemoglobin oxygen saturation (84 ± 9.5%) and by blood gas PaO₂ (44.5 ± 5.8 mm Hg). It was concluded that the HD provided more consistent results than the LD, but induced transient hypoxemia. Tolazoline shortened the recovery time in llamas receiving the HD.     

Effects of sun compass error on spatial search by Clark's nutcrackers

Animals employ compasses during navigation, but little attention has been paid to how accuracy is maintained in the face of compass error, which is inevitable in biological systems. The use of multiple landmarks may minimize the effect of compass error. We allowed Clark's nutcrackers to cache seeds in an outdoor aviary with either one or four landmarks present, and subsequently subjected them to small clock‐shifts mimicking the effects of compass error. As predicted, the results showed a significant decrease in search accuracy following the clock‐shift when one landmark was present but not when four landmarks were present. These results support that nutcrackers encode information from the sun as well as terrestrial landmarks, and these spatial cues are used in a flexible manner. Overall, our results are important as they support the hypothesis that multiple landmarks may be used during situations where the sun compass has even a small amount of error.     

Immunohistochemical Localization of Oestrogen Receptors α and β, Progesterone Receptor and Aromatase in the Equine Placenta

The functions of placental oestrogens during equine pregnancy are still unclear. Yet, they may act predominantly as local regulators of growth and differentiation in the microplacentomes. Thus, expression patterns of oestrogen receptors (ERs) α and β were investigated in the microcotyledonary placenta from pregnant mares at 110, 121, 179, 199 and 309 days of gestation by immunohistochemistry. In microplacentomes, both the ER isoforms were detected in trophoblast (T) cells, chorionic villous stroma (FS), microcaruncular epithelium (ME) and microcaruncular stroma (MS). Proportions of positive cells were 38–91% (T), 11–41% (FS), 55–89% (ME), 17–51% (MS) for ERα and 66–76% (T), 21–37% (FS), 41–68% (ME) and 24–55% (MS) for ERβ. Between days 110 and 199, proportions of cells positive for progesterone receptor (PR) varied between 19% and 62% (T), 3% and 50% (CS), 15% and 46% (ME), and 4% and 33% (MS). At day 309, PR was virtually absent in T, CS and ME (percentages < 0.1), whereas in MS 14.3% of cells were still positive. The expression of ERs and PR in equine microplacentomes gives evidence for a role of placental steroids as regulators of placental growth, differentiation and function. The detection of ERα, ERβ and PR in foetal and maternal vascular tissue suggests that placental steroids are also involved in the control of placental angiogenesis and /or vascular functions. The co-localization of ERs with aromatase in T suggests auto- or intracrine functions of oestrogens in this cell type.     

Histological and Immunohistochemical Characterization of Equine Anovulatory Haemorrhagic Follicles (AHFs)

Anovulatory haemorrhagic follicles (AHFs) are often the reason for ovulation failure in the mare. As the underlying factors that lead to AHF development are not well understood, it was of interest to investigate the vascularization of AHFs compared with normal follicles and corpora lutea (controls). In the present study, the ovarian cell populations investigated immunohistochemically included granulosa and luteal cells as well as various vascular structures. None of these cell types showed differences in the expression of vascular endothelial growth factor A (VEGF-A) between control ovaries containing normal follicles and corpora lutea and ovaries with AHFs. In contrast, a considerable reduction in the proportion of Flk-1-expressing cells, together with a decreased intensity of staining, was apparent in the AHFs. This greatly reduced expression of Flk-1 in the luteinized cells and the vascular structures of AHFs may lead to a distinct decrease in the potential pro-angiogenic activity of VEGF-A in these structures compared with the situation in normal follicles and corpora lutea. Furthermore, the authors suspect that the distinct expression of angiopoietin2 and VEGF-A seen in the cells within the inner fibrous layers of the AHFs was caused by hypoxia resulting from deficient vascularization, as suggested by the irregularity of the capillaries present in the luteinized wall of the AHF. In addition, whereas LH-receptor (LH-R) expression occurred uniformly in all stages of development of the corpora lutea in normal control ovaries, there was highly variable labelling for LH-R in all the AHFs examined, thereby indicating a possible numerical deficiency of LH-receptors in AHFs. The authors concluded that, despite the apparent expression of sufficient VEGF-A in the AHFs allows ovulation and corpus luteum formation, a relative lack of receptor, Flk-1, effects the pro-angiogenic activity of VEGF-A which could be a reason for ovulation failure associated with AHF formation.     

The Development and Application of the Modern Reproductive Technologies to Horse Breeding

Although the horse was probably the first animal to experience and benefit from artificial insemination, it trailed the field somewhat with regard to the application of embryo transfer and other oocyte and embryo-related modern breeding technologies. But with a late run it is now back in mid-field and gaining fast on the other large domestic species in the application of the many technological advances of the past 20 years to sound breeding practice. Improvements in extenders and cryoprotectants have resulted in a veritable upsurge in the transport and insemination of cooled and frozen stallion semen, and parallel improvements in ovulation induction and synchrony, exogenous gonadotrophic stimulation of multiple fertile ovulations and simplified, more efficient methods for non-surgical transfer of embryos to recipient mares, coupled with relaxation of breed society registration restrictions, have together contributed to a similar upsurge in the application of embryo transfer to all breeds and athletic types of horses worldwide, with the continuing and notable exception of the Thoroughbred. Although conventional in vitro fertilization remains something of an unjumped fence in equids, other modern breeding technologies like hysteroscopic low-dose insemination, fluorescence-activated sex sorting of stallion spermatozoa, between-species embryo transfer, embryo freezing and bisection, transvaginal ultrasound-guided oocyte collection, intracytoplasmic sperm injection for fertilization (ICSI), gamete intrafallopian transfer (GIFT) and now nuclear transfer (cloning), have all been applied to equids with encouraging success. Cloning, especially, holds enormous promise for the Sporthorse industry to re-create champion geldings in stallion form for breeding purposes.     

In Vitro Pullout Strength of Screws Inserted in Adult Equine Third Metacarpal Bone After Overdrilling a 4.5-mm Threaded Insertion Hole

Objective—To determine and compare the in vitro pullout strength of 5.5-mm cortical versus 6.5-mm cancellous bone screws inserted in the diaphysis and metaphysis of adult equine third metacarpal (MCIII) bones, in threaded 4.5-mm cortical bone screw insertion holes that were then overdrilled with a 4.5-mm drill bit to provide information relevant to the selection of a replacement screw if a 4.5-mm cortical screw is stripped. Study Design—In vitro pullout tests of 5.5-mm cortical and 6.5-mm cancellous screws in equine MCIII bones. Sample Population—Two independent cadaver studies each consisting of 14 adult equine MCIII bones. Methods—Two 4.5-mm cortical screws were placed either in the middiaphysis (study 1) or distal metaphysis (study 2) of MCIII bones. The holes were then overdrilled with a 4.5-mm drill bit and had either a 5.5-mm cortical or a 6.5-mm cancellous screw inserted; screw pullout tests were performed at a rate of 0.04 mm/second until screw or bone failure occurred. Results—In diaphyseal bone, the screws failed in all tests. Tensile breaking strength for 5.5-mm cortical screws (997.5 ± 49.3 kg) and 6.5-mm cancellous screws (931.6 ± 19.5 kg) was not significantly different. In metaphyseal bone, the bone failed in all tests. The holding power for 6.5-mm cancellous screws (39.1 ± 4.9 kg/mm) was significantly greater than 5.5-mm cortical screws (23.5 ± 3.5 kg/mm) in the metaphysis. There was no difference in the tensile breaking strength of screws in the diaphysis between proximal and distal screw holes; however, the holding power was significantly greater in the distal, compared with the proximal, metaphyseal holes. Conclusions—Although tensile breaking strength was not different between 5.5-mm cortical and 6.5-mm cancellous screws in middiaphyseal cortical bone, holding power of 6.5-mm cancellous screws was greater than 5.5-mm cortical screws in metaphyseal bone of adult horses. Clinical Relevance—If a 4.5-mm cortical bone screw strips in MCIII diaphyseal bone of adult horses, either a 5.5-mm cortical or 6.5-mm cancellous screw, however, would have equivalent pullout strengths. A 6.5-mm cancellous screw, however, would provide greater holding power than a 5.5-mm cortical screw in metaphyseal bone.     

Biomechanical Comparison of Six Suture Configurations Using a Large Diameter Polyester Prosthesis in the Muscular Process of the Equine Arytenoid Cartilage

Objective— To compare biomechanical properties of 6 suture configurations using a large diameter polyester prosthesis in the muscular process (MP) of the arytenoid cartilage and to determine failure mode.
Study Design— Experimental study.
Sample Population— Cadaveric equine larynges (n=121).
Methods— Suture configurations (4 single, 2 double) were inserted, and then constructs were tested in a single-cycle to failure at a 100 mm/min distraction. Load deformation curves were generated to assess the biomechanical properties of each construct. A 1-way ANOVA was used to compare the mean differences in construct failure force, cricoarytenoid joint (CAJ) disarticulation force, and energy stored at failure. A 2-sample t-test was used to compare single versus double suture patterns and a Fisher's exact test was used to compare failure mode.
Results— Both construct and CAJ failure force were significantly greater ( P <.05) for double suture patterns compared with single suture patterns; however, there were no significant differences in energy stored at construct failure. Failure at the MP accounted for ≥50% of construct failures for 3 of the single suture patterns and 1 of the double suture patterns tested. The remaining 2 patterns had an increased frequency of clamp failures as well as failure of the cricoid cartilage.
Conclusions— Sutures that sufficiently engage the spine of the MP alone or in conjunction with a second suture were found to be biomechanically superior.
Clinical Relevance— Engaging the spine of the MP appears to result in the most biomechanically sound laryngeal construct.     

Immunohistochemical Localization of Vascular Endothelial Growth Factor (VEGF) and its Two Receptors (Flt-I and KDR) in the Endometrium and Placenta of the Mare During the Oestrous Cycle and Pregnancy

Polyclonal antisera for vascular endothelial growth factor (VEGF) and its two main receptor molecules, VEGF-I (Flt) and VEGF-II (KDR), were used in a conventional immunocytochemical staining method to localize these angiogenic ligand molecules in the endometrium and placenta of the mare during the oestrous cycle and pregnancy. The anti-VEGF and anti-Flt sera both labelled the lumenal and glandular epithelia of the endometrium throughout the oestrous cycle and both the invasive trophoblast cells of the endometrial cups and the non-invasive trophoblast of the allantochorion in pregnancy. The anti-KDR serum likewise stained the maternal and foetal epithelial layers during the oestrous cycle and pregnancy and it also labelled fibroblast-like cells in the endometrial and allantoic stromas and the endothelium of foetal and maternal capillaries. The results demonstrated that constant supplies of the principal vasculogenic and angiogenic factor, VEGF, and its two major receptors, Flt and KDR, are available on both the maternal and foetal sides of the placental barrier throughout gestation in the mare. They are presumed to facilitate the continuing development of the extensive foetal and maternal capillary networks that are such prominent features within the microplacentomes of the diffuse, epitheliochorial equine placenta.