Evaluation of the sublingual microcirculation with sidestream dark field video microscopy in horses anesthetized for an elective procedure or intestinal surgery.

OBJECTIVE: To compare the sublingual microcirculation between healthy horses anesthetized for elective procedures and horses with colic anesthetized for abdominal surgery and to determine the effect of mean arterial blood pressure (MAP) on the microcirculation. ANIMALS: 9 horses in the elective group and 8 horses in the colic group. PROCEDURES: Sublingual microcirculation was assessed with sidestream dark field video microscopy. Videos were captured at 3 time points during anesthesia. Recorded microvasculature parameters were De Backer score (DBS), total density of perfused vessels (PVD) and small vessels (PVD-S), total proportion of perfused vessels (PPV) and small vessels (PPV-S), vascular flow index (MFI), and heterogeneity index (HI). Blood pressure during hypotensive (MAP < 60 mm Hg) and normotensive (MAP ≥ 60 mm Hg) episodes was also recorded. RESULTS: During normotensive episodes, the elective group had significantly better PPV and PPV-S versus the colic group (median PPV, 76% vs 50%; median PPV-S, 73% vs 51%). In both groups, PPV decreased during anesthesia (elective group, -29%; colic group, -16%) but significantly improved in the elective group 15 minutes before the end of anesthesia (59%). During hypotensive episodes, PVD-S was better preserved in the colic group (11.1 vs 3.8 mm/mm2). No differences were identified for the microcirculatory parameters between normo- and hypotensive episodes in the colic group. CONCLUSIONS AND CLINICAL RELEVANCE: Sublingual microcirculation was better preserved in healthy horses anesthetized for elective procedures than in horses with colic anesthetized for abdominal surgery despite resuscitation maneuvers. Results indicated that the macrocirculation and microcirculation in critically ill horses may be independent.

Evaluation of jejunal microvasculature of healthy anesthetized dogs with sidestream dark field video microscopy.

OBJECTIVE: To determine the feasibility of sidestream dark field (SDF) video microscopy for the evaluation of the jejunal microvasculature of healthy dogs. ANIMALS: 30 healthy sexually intact female shelter dogs anesthetized for ovariohysterectomy. PROCEDURES: Preoperative physical and clinicopathologic assessments were performed to confirm health status. Then healthy dogs were anesthetized, and the abdomen was incised at the ventral midline for ovariohysterectomy and jejunal microvasculature evaluation. An SDF video microscope imaged the microvasculature of 2 sites of a portion of the jejunum, and recorded videos were analyzed with software capable of quantitating parameters of microvascular health. Macrovascular parameters (heart rate, respiratory rate, and hemoglobin oxygen saturation) were also recorded during anesthesia. RESULTS: Quantified jejunal microvascular parameters included valid microvascular density (mean ± SD, 251.72 ± 97.10 µm/mm), RBC-filling percentage (66.96 ± 8.00%), RBC column width (7.11 ± 0.72 µm), and perfused boundary region (2.17 ± 0.42 µm). The perfused boundary region and RBC-filling percentage had a significant negative correlation. Strong to weak positive correlations were noted among the perfused boundary regions of small-, medium-, and large-sized microvessels. No significant correlations were identified between microvascular parameters and age, body weight, preoperative clinicopathologic results, or macrovascular parameters. CONCLUSIONS AND CLINICAL RELEVANCE: Interrogation of the jejunal microvasculature of healthy dogs with SDF video microscopy was feasible. Results of this study indicated that SDF video microscopy is worth additional investigation, including interrogation of diseased small intestine in dogs.

Mechanical Coupling between Endoderm Invagination and Axis Extension in Drosophila.

How genetic programs generate cell-intrinsic forces to shape embryos is actively studied, but less so how tissue-scale physical forces impact morphogenesis. Here we address the role of the latter during axis extension, using Drosophila germband extension (GBE) as a model. We found previously that cells elongate in the anteroposterior (AP) axis in the extending germband, suggesting that an extrinsic tensile force contributed to body axis extension. Here we further characterized the AP cell elongation patterns during GBE, by tracking cells and quantifying their apical cell deformation over time. AP cell elongation forms a gradient culminating at the posterior of the embryo, consistent with an AP-oriented tensile force propagating from there. To identify the morphogenetic movements that could be the source of this extrinsic force, we mapped gastrulation movements temporally using light sheet microscopy to image whole Drosophila embryos. We found that both mesoderm and endoderm invaginations are synchronous with the onset of GBE. The AP cell elongation gradient remains when mesoderm invagination is blocked but is abolished in the absence of endoderm invagination. This suggested that endoderm invagination is the source of the tensile force. We next looked for evidence of this force in a simplified system without polarized cell intercalation, in acellular embryos. Using Particle Image Velocimetry, we identify posteriorwards Myosin II flows towards the presumptive posterior endoderm, which still undergoes apical constriction in acellular embryos as in wildtype. We probed this posterior region using laser ablation and showed that tension is increased in the AP orientation, compared to dorsoventral orientation or to either orientations more anteriorly in the embryo. We propose that apical constriction leading to endoderm invagination is the source of the extrinsic force contributing to germband extension. This highlights the importance of physical interactions between tissues during morphogenesis.

Video Telescope Operating Microscopy.

Exotic pet veterinarians frequently have to operate on small animals, and magnification is commonly used. Existing endoscopy equipment can be used with a mechanical arm and telescope to enable video telescope operating microscopy. The additional equipment items and their specifics are described, and several case examples are provided.

Assessment of microcirculatory perfusion in healthy anesthetized cats undergoing ovariohysterectomy using sidestream dark field microscopy.

OBJECTIVE: To: (1) determine the feasibility of using sidestream dark field microscopy (SDM) to measure microcirculatory parameters in healthy, anesthetized cats and (2) determine if surgical tissue manipulation and anesthesia time alter these parameters during ovariohysterectomy. DESIGN: Prospective observational study. SETTING: University teaching hospital. ANIMALS: Eighteen healthy female cats. INTERVENTIONS: Sublingual mucosa microcirculatory videos were obtained under general anesthesia preoperatively, intraoperatively, and postoperatively using an SDM device in healthy cats presenting for ovariohysterectomy. At each video acquisition point, macrovascular parameters (heart rate, blood pressure, pulse oximetry, end-tidal CO2) were recorded. Vascular analysis software was used to calculate standard microcirculatory parameters. Multivariate analysis was performed to compare microvascular and macrovascular parameters, as well as correlation with the effect of surgical manipulation and time under anesthesia. MEASUREMENTS AND MAIN RESULTS: Twelve of 18 cats were included in final video analysis; 6 were removed for poor video quality. Values for total vessel density (TVD, 47.7 ± 8.39 mm/mm(2)), proportion of perfused vessels (PPV, 88.2 ± 5.95%), perfused vessel density (PVD, 43.0 ± 9.00 mm/mm(2)), microcirculatory flow index (MFI, 2.33 ± 0.33) were determined preoperatively. There were no significant changes in TVD, PPV, and PVD across intervention points. The MFI increased significantly from preoperative to intra- and postoperative data collection points. No correlation between microcirculatory parameters and length of anesthesia or macrocirculatory values was found. CONCLUSIONS AND CLINICAL RELEVANCE: This study demonstrated that SDM can be utilized to obtain sublingual microvascular parameters in healthy, anesthetized cats. Limitations include difficulty in obtaining high quality images, presumed need for general anesthesia, and need for off-line video analysis. This technology has potential as a tool in experimental and clinical monitoring of microcirculatory changes in felines.

Evaluation of a rapid bedside scoring system for microcirculation videos acquired from dogs.

OBJECTIVE: To appraise a novel scoring system (Bedside Evaluation of Microcirculation [BEM]) to provide qualitative and semiquantitative assessment of canine microcirculatory videos generated by sidestream dark field imaging. DESIGN: Prospective observational study. SETTING: University teaching hospital. ANIMALS: No animals were used in this study. Twenty microcirculatory videos (>20 s in length) acquired from the mucosa adjacent to the upper canine tooth of dogs were selected from a database of sidestream dark field microcirculatory videos with available current standard analysis (CSA). INTERVENTION: Three observers were trained to evaluate 5 video quality parameters (stability, content, illumination, focus, and pressure) and four perfusion parameters (total vessel density [TVD], capillary vessel density [CVD], perfused vessel density [PVD] and microvascular flow index [MFI]). Quality parameters were scored (excellent [0], sufficient [1], and insufficient [2]) similar to CSA recommendations. Each perfusion parameter was subjectively scored (1 lowest - 5 highest) using sample clips from the training video for comparison. Videos passed quality analysis if no parameter was scored insufficient. Repeatability and reproducibility were evaluated by assessing all films in a random order three times daily for 3 days. Strength of correlation of BEM with CSA for both qualitative and semiquantitative parameters was assessed. MEASUREMENTS AND MAIN RESULTS: The qualitative evaluation pass/fail assessment matched CSA 86% of the time with individual observer agreements of 84-88%. Agreement with CSA did not change significantly over the study period (84%, 88%, and 84% on days 1, 2, and 3, respectively). No significant correlations were demonstrated between any BEM perfusion parameter and the corresponding CSA values. CONCLUSIONS: Rapid bedside assessment of microcirculatory video quality can be achieved. However, semiquantitative analysis by BEM demonstrated a lack of correlation with CSA for any of the perfusion parameters assessed.

Time-lapse videomicrographic observations of blastocyst hatching in cattle.

Morphological changes of cultured bovine blastocysts during hatching were observed using time-lapse videomicrography in order to investigate the patterns of the hatching process that occurred in the blastocysts and to determine whether the hatching patterns differed between blastocysts developed from fresh and cryopreserved embryos. Compacted morulae (CMs) were collected from superovulation-treated Japanese Black and Holstein dairy cattle and cultured in a medium in a CO(2) culture chamber equipped with an inverted microscope at 38.5 C. Images of resultant blastocysts during the period from blastocoel formation to completion of hatching were taken at 4-sec intervals by a CCD color camera connected to an inverted microscope and recorded by a time-lapse video cassette recorder. In blastocysts developed from fresh CMs, hatching was found to begin with protrusion of trophectoderm cells from zonae pellucidae at the expanded stage. Protrusion of the cells occurred in any site of the trophectoderm. After protrusion, a large or small slit was formed in the zona pellucida in all blastocysts as a result of blastocyst expansion or enlargement of the protrusion. Then, blastocysts completely escaped from the zona pellucida through the slit in the state of expansion. From these findings, the hatching patterns of cattle blastocysts could be classified into 5 types. In blastocysts developed from frozen-thawed CMs, the hatching pattern and length of time needed for hatching are similar to those in blastocysts developed from fresh CMs. In addition, the pregnancy rate of recipients following transfer of frozen-thawed CMs (52.4%) did not differ from that of recipients following transfer with fresh CMs (58.3%). These results suggested that the quality of frozen-thawed cattle embryos is comparable to that of fresh embryos and that there could be a relationship between the hatching pattern of blastocysts and the viability of embryos after transfer.

Evaluating the impacts of osmotic and oxidative stress on common carp (Cyprinus carpio, L.) sperm caused by cryopreservation techniques.

Cryopreservation causes osmotic changes and oxidative damage that have sublethal and lethal effects on spermatozoa. We examined these osmotic and oxidative effects on common carp spermatozoa motility; membrane integrity; levels of thiobarbituric-acid-reactive substance (TBARS) and carbonyl groups (CP); and the activity of superoxide dismutase (SOD), glutathione reductase, and glutathione peroxidase (GPx). Sperm was diluted in dimethyl sulfoxide (DMSO) and ethylene glycol-based extenders, followed by equilibration, freezing, and thawing. Equilibration in DMSO extender resulted in a significant reduction of spermatozoa motility, but motility was induced in those spermatozoa following dilution with saline buffer, which usually inhibits undiluted spermatozoa motility. Spermatozoa velocity and membrane integrity decreased with both extenders following freezing and thawing. No significant difference in levels of TBARS or CP, or in SOD activity, was seen in samples equilibrated with either extender. The freeze/thaw process induced significantly higher levels of TBARS, CP, and GPx activity, but did not affect the level of SOD. Glutathione reductase activity was inhibited in samples exposed to DMSO extender. Ethylene glycol should be considered a preferred cryoprotective agent for common carp spermatozoa to reduce osmotic and oxidative stress during cryopreservation.

Measurements of microvascular perfusion in healthy anesthetized dogs using orthogonal polarization spectral imaging.

OBJECTIVE: To determine normal microvascular assessment parameters for healthy, anesthetized dogs. DESIGN: Prospective investigational descriptive study. SETTING: University Teaching Hospital. ANIMALS: Fifteen client-owned, systemically healthy dogs that were undergoing general anesthesia for an elective procedure. INTERVENTIONS: A sidestream dark-field videomicroscope probe was placed in the mouth at the mucogingival junction above the canine tooth and 3 video recordings of the microcirculation were made for later analysis by 2 independent, blinded reviewers. MEASUREMENTS AND MAIN RESULTS: The videos were analyzed to determine the total vessel density, proportion of perfused vessels, microcirculatory flow index, and perfused vessel density. A range of values for these indices were obtained and reported. CONCLUSIONS: The microcirculation of normal dogs is readily observable using the videomicroscope and recorded video segments can be used to determine microcirculatory measurements. These values may prove useful for comparison in future studies that examine canine microcirculatory parameters.

A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo.

Much of the current knowledge of human cardiovascular pathologies and treatment strategies has been gained from understanding the cardiac physiologies and functions in small animal models, such as mice, rats, and zebrafish. In this paper, we present the development of a high-frame-rate duplex ultrasound biomicroscopy (UBM) capable of B-mode imaging and pulsed-wave (PW) Doppler measurement for in vivo cardiovascular investigation in small animals. A frame rate of 200 frames per second (fps) was accomplished at a view of 5 mm x 8 mm, using a novel high-speed sector probe and specially designed lightweight transducers. In a reduced lateral view of 1.2 mm, a frame rate of 400 fps was achieved to examine more detailed cardiac motion. The UBM utilized transducers with different center frequencies (40-75 MHz) and geometries, which made it useful for various applications in small animal cardiac imaging. The highest spatial resolution the UBM achieved was 25 microm x 56 microm. In addition, the image-guided PW Doppler implemented in the UBM demonstrated the detection of the velocity of a moving wire as low as 0.1 mm/s, and flow in a polyimide tube as small as 200 microm in diameter. Furthermore, the UBM achieved a 15- microV minimal detectable signal and a 60-dB dynamic range using a low-cost PCB-based design. Finally, sample in vivo cardiac images of mouse and zebrafish hearts were given. These results showed that the UBM integrated with B-mode and PW Doppler is useful to investigate the pathophysiological mechanism in the cardiovascular studies.

Ticks, Amblyomma rotundatum (Acari: ixodidae), on toads, Chaunus schneideri and Chaunus granulosus (Anura: bufonidae), in northern Argentina.

This communication provides notes on 2 species of toads, Chaunus schneideri and Chaunus granulosus, infested with ixodid ticks, Amblyomma rotundatum, from the provinces of Corrientes and Formosa in northern Argentina. Chaunus schneideri is a new amphibian host record for A. rotundatum, a species previously reported to parasitize other anurans and also reptiles. We examined 74 ticks on 5 toads. All ticks were A. rotundatum; all adults were females, and all developmental stages were randomly attached to host body parts. Ticks remained attached to one of the toads for from 7 to 17 days after the host was captured. One toad, encumbered with 33 ticks, was moribund when found and died shortly thereafter.

The temporal relationship between oocyte maturation and early fertilisation events in relation to the pre-ovulatory LH peak and preimplantation embryo development in red deer (Cervus elaphus).

The temporal relationships among oocyte maturation, gamete transport and fertilisation following the pre-ovulatory luteinsing hormone surge in red deer were established; and secondly, early preimplantation development to the blastocyst stage in relation to the onset of oestrus was determined for red deer. In the first series of observations, oestrus was synchronised in April (N=22), for the fixed time recovery of gametes from 0 to 36 h after the estimated pre-ovulatory LH peak. Matings were observed and the time of the LH peak was determined from the retrospective analysis of blood plasma collected at 3h intervals. Gametes were recovered surgically and the meiotic status of follicular and ovulated oocytes assessed. Spermatozoa were recovered from the oviduct and their motility analysed by videomicroscopy. Nineteen of 22 hinds exhibited a pre-ovulatory LH surge and were observed to mate. Oocyte metaphase I occurred between 11 and 18 h, and metaphase II was completed within the follicle between 20 and 25 h following the pre-ovulatory LH peak. Fertilised ova were recovered from 30 to 36 h in both the ampulla and isthmic portions of the oviduct. Motile spermatozoa were first recovered from the isthmus and the ampulla at 13 and 21 h, respectively, after the LH peak. Hyperactive spermatozoa were observed in both the isthmus and the ampulla flushings but only from the eight hinds that had ovulated. In the second series of observations, 16 mature hinds were synchronised and allocated to groups for embryo collection on days 3, 5 and 7 after oestrus. Eight embryos were recovered; an 8-cell at 90 h, 3 morulae at 137, 138 and 186 h, and 4 blastocysts at 180, 182 and 190 h post-mating. Blastocysts were only recovered from the uterine horns and the mean+/-S.E.M. number of nuclei per blastocyst was 93.5+/-10.0 with a range of 66-114 cells. The results of this study will improve the application of assisted reproductive technologies to red deer as they indicate that oocyte maturation, fertilisation and early embryonic development of the red deer is similar to other domestic ruminants with the exception that the red deer embryo enters the uterus at the blastocyst stage.

Long-term observation of pial microcirculatory parameters using an implanted cranial window method in the rat.

The aim of this study was to confirm whether our improved closed cranial window (CCW) method could be used for long-term microscopical observation of pial microcirculation intravitally in the rat. We investigated chronological changes in three microcirculatory parameters: permeability of blood-brain barrier, leukocyte behavior, and plasma velocities in the pial venules, immediately after implantation (control group) and at one and four weeks after implantation in different age-matched rats (implanted group). No extravasation of sodium fluorescein from pial venules was confirmed in any observation period. The number of endothelial-adhering leukocytes in the implanted group kept within the physiological range, being similar to those of the control group. The velocities of fluorescent microspheres flowing in pial venules showed no noticeable changes between the two groups. These findings suggest that our CCW method allows long-term observation of the pial microcirculation without any pathophysiological changes in the evaluated parameters up to four weeks after the implantation.

Tritrichomonas foetus damages bovine oocytes in vitro.

Tritrichomonas foetus is an extracellular parasite of the urogenital tract in cattle. It causes infertility and abortion, but there is no documented information on the susceptibility of bovine oocytes to the parasite, except by one article that claimed no effects of T. foetus on oocytes or embryos. The aim of the present study was to study the effects provoked by T. foetus when in interaction with bovine oocytes. Oocytes were obtained from cow ovaries and divided into two groups: (1) one group contained cumulus cells, whereas (2) a second group was denuded from these cells. Light microscopy, video microscopy and scanning electron microscopy (SEM) revealed that exposure of oocytes to T. foetus caused rapid adhesion of the trichomonads to cumulus cells and to the zona pellucida (ZP). Motile parasites were observed for 12 h. The ZP was completely damaged, and the parasites were able to infiltrate beneath the ZP and reached the oocytes directly when the oocytes were denuded of the cumulus cells. Both the oocytes and the cumulus cells exhibited morphological characteristics compatible with apoptosis after interaction with T. foetus, such as chromatin condensation, the presence of several cytoplasmic vacuoles, with intact cellular membranes and organelles. The results from this study demonstrate that when a large number of T. foetus interacts with oocytes in vitro damage and apoptosis are provoked in the cow's reproductive cells. The behavior of this parasite as one of the causes of cattle infertility is discussed.

Measurement of eye-gaze in chimpanzees (Pan troglodytes).

Gaze cues are used as an index of social cognition in primates, yet the sensitivity to different forms of gaze, and consequently the cues required to test gaze-following abilities remain understudied. Whereas the eye is attributed special signal value in humans, the camouflaged ocular morphology of non-human primates has led to the consensus that head orientation may be a more salient cue. This study presents the first documentation of the surface eye movements of the chimpanzee, Pan troglodytes, in order to determine the behavioral forms of eye-gaze and their saliency as signals, document their functional variation, and address the signal value of the eyes distinct from head orientation. Movements of the eye were identified as Scan (continuous movement), Glance (a single movement <1 sec), or Fixate (no movement). Scans, glances, and fixations were reliably detected by humans during live observation and from video (Cohen's kappa over 0.70) and, therefore, are likely also to be detected by conspecifics. Eye-gaze comprised a nonunitary measure of visual attention, reflecting the attentional task demands of different activities. Specifically, chimpanzees spent significantly more time scanning while feeding and resting, than grooming, F(2,28) = 10.23, P<0.001, and spent significantly more time fixating while grooming, than feeding or resting, F(2,28) = 7.52, P<0.01. Further, eye-gaze was often incongruent with head movement, varying significantly with the form of eye-gaze: incongruence was found during 12-21% of fixations, during 42-49% of scans, and during 70-100% of glances, F(2,16) = 30.17, P<0.001. These findings provide the basis for discrimination of the adaptive significance of gaze-processing abilities with emphasis on sensitivity to eye-gaze distinct from head orientation. If we are to continue exploring gaze-processing abilities in primates, then we need greater consideration of the precise nature of the signals themselves. Here we present evidence for special consideration of the eyes as a salient signal in P. troglodytes.

Development of a novel CASA system based on open source software for characterization of zebrafish sperm motility parameters.

Although computer-assisted sperm analysis (CASA) outperforms manual techniques, many investigators rely on non-automated analysis due to the high cost of commercial options. In this study, we have written and validated a free CASA software primarily for analysis of fish sperm. This software is a plugin for the free National Institutes of Health software ImageJ and is available with documentation at . That it is open source makes possible external validation, should improve quality control and enhance the comparative value of data obtained among laboratories. In addition, we have improved upon the traditional velocity straight line (VSL) algorithm, eliminating inaccurate characterization of highly curved fish sperm paths. Using this system, the motion of zebrafish (Danio rerio) sperm was characterized relative to time post-activation and the impact of acquisition conditions upon data analysis determined. There were decreases in velocity and path straightness (STR), but not linearity (LIN), relative to time. From 30 to 300 frames/s, frame rate significantly affected curvilinear velocity (VCL) and STR measurements. Sperm density in the field of view did not affect any measured parameter. There was significant inter-male variation for VCL, VSL, velocity average path (VAP), percent motility, path character (STR, LIN), and duration of motility. Furthermore, relative sperm output (a measure reflecting both semen volume and concentration) was positively correlated to percent motility. For all motion parameters measured (except duration), the average CV was < or =10%, comparable to values obtained using commercial systems.

Redescription of Ascocotyle (Ascocotyle) felippei Travassos, 1928 (Digenea: Heterophiydae) with new synonymies.

The heterophyid trematode Ascocotyle (Ascocotyle) felippei Travassos, 1928, is redescribed and new data on its life cycle are provided, based on types and metacercariae found in the heart bulb and gills of naturally infected guppies, Poecilia vivipara (new fish intermediate host), from a coastal lagoon in Rio de Janeiro, Brazil. Examination of the type and all voucher specimens of A. (A.) felippei collected by Travassos in the type host and locality in Brazil has shown that they possess only 32 (16 + 16) circumoral spines, rather than 36 (18 + 18) spines as previously reported. Based on the identical number and arrangement of circumoral spines, shape of the body, the presence of a long preoral lobe and posterior muscular prolongation of the oral sucker, short and wide ceca, a simple gonotyl lacking refractile bodies, and the site of infection of metacercariae (predominantly heart bulb), A. (A.) puertoricensis Price, 1932 and A. (A.) tenuicollis Price, 1935, are proposed as new synonyms of A. (A.) felippei.

Site-specific adaptive remodeling of Greyhound metacarpal cortical bone subjected to asymmetrical cyclic loading.

OBJECTIVE: To quantify geometric, inertial, and histomorphometric properties at the mid-diaphyseal level of left and right metacarpal bones (MCB) of racing Greyhounds. SAMPLE POPULATION: MCB from 7 racing Greyhounds euthanatized for reasons unrelated to MCB abnormalities. PROCEDURES: Mid-diaphyseal transverse sections of left and right MCB were stained with H&E or microradiographed. Images of stained sections were digitized, and cross-sectional area, cortical area, and maximum and minimum area moments of inertia of each bone were determined. Histomorphometric data (osteonal density, osteonal birefringence, and endosteal new lamellar bone thickness) were collected in 4 quadrants (dorsal, palmar, lateral, medial). Values were compared between limbs and among bones and quadrants. RESULTS: Cross-sectional area, cortical area, and maximum and minimum moments of inertia of left MCB-IV and -V were significantly greater, compared with contralateral bones. Overall osteonal densities in the dorsal quadrants of left MCB were greater, compared with lateral and medial quadrants. Also, percentage of birefringent osteons was significantly greater in the dorsal quadrant of left MCB-III, -IV, and -V, compared with the palmar quadrant. Thickness of new endosteal lamellar bone was not significantly influenced by limb, bone, or quadrant. CONCLUSIONS AND CLINICAL RELEVANCE: Increased cortical thickness and geometric properties of left MCB-IV and -V of Greyhounds, together with altered turnover and orientation of osteons in the dorsal quadrants of left MCB, are site-specific adaptive responses associated with asymmetric cyclic loading as a result of racing on circular tracks. Site-specific adaptive remodeling may be important in the etiopathogenesis of fatigue fractures in racing Greyhounds.

Observations on the feeding behaviour of Paratrichodorus anemones in relation to tobravirus transmission.

Feeding by P. anemones, an efficient vector of tobacco rattle virus (TRV), was investigated by video-enhanced interference light microscopy. Four stages were observed after transfer of individual nematodes, extracted from soil, to Nicotiana tabacum seedling roots in agar: i) acclimatisation; ii) approach and scrutiny; iii) preparation; and iv) feeding. Prior to commencement of stage 'iv' about 4 cells perforated by rapid onchiostyle thrusting remained alive, each having been almost immediately abandoned by the nematode. During the stage iv) approximately 5% of perforated cells remained alive. Feeding on individual cells was similar to that previously reported for Trichodorus similis: cells from which cytoplasm was ingested after a prolonged period of salivation were invariably killed, with adjacent cells being unaffected. During feeding a number of cells perforated but soon afterwards abandoned by P. anemones remained alive, providing an effective pathway for successful transmission of TRV to plants by the nematode.

Factor XIIIa positive dendritic cells are a major accessory cell population in the elicitation phase of DNCB-induced contact hypersensitivity.

The phenotypes and distribution of accessory cells in the ear skin of lambs during the elicitation phase of dinitrochlorobenzene (DNCB)-induced contact hypersensitivity (CHS) were examined using indirect immunoperoxidase histochemistry (ABC method), and a panel of antibodies. Thirty lambs, between 21 and 26 weeks of age, were divided into groups of 10. The shaved right ear of one group was treated with DNCB. Two weeks later this group was challenged with DNCB. One group was treated with the vehicle alone and the remaining group was left untreated. The lambs were slaughtered 48 h after challenge, and tissue specimens were collected from the ears of the three groups. Factor XIIIa+ (FXIIIa+) cells were prominent in the superficial dermis and showed predominantly a perivascular and subepidermal distribution. The other markers were less prominent, and whereas CD1+ cells and CD68+ cells showed a reaction pattern similar to the FXIIIa+ cells, CD14+ cells were found scattered predominantly in the deep dermis. There appeared to be an increase in FXIIIa+ cells, CD1+ cells, and CD68+ cells in the dermis of the DNCB-treated lambs 48 h after challenge. Only CD1+ cells were detected in epidermis of normal controls, and these cells appeared to be decreased in number in the two treated groups. Computer-assisted morphometric analysis was used to estimate the relative presence of the accessory cell subpopulations in the superficial and deep dermis and the entire dermis. A statistical analysis of the relative area of immunostaining showed a significantly increased presence of FXIIIa+ cells and CD68+ cells in the dermis of the DNCB-treated lambs 48 h after challenge. Interestingly, FXIIIa+ cells and CD68+ cells were also significantly increased in the vehicle treated group compared with untreated controls. We found no significant difference in the presence of CD1+ cells or CD14+ cells in the DNCB treated group compared with the controls. The study showed that FXIIIa+ DDC are the major accessory cell population in normal ear skin of lambs and the major responsive population during the elicitation phase of CHS. The lack of response in the CD1+ cell population suggests a less prominent role for the LC-related DC in the skin during the elicitation phase.