Hexosamine concentration, DNA concentration, and [35S]sulfate incorporation for articular cartilage obtained from various sites in the metacarpophalangeal and carpal joints of horses were measured. The same measurements were made on the repair tissue filling full-thickness articular defects in the intermediate carpal bone and on cartilage surrounding partial-thickness defects 6 weeks after the defects were created arthroscopically. Cellularity (measured as DNA concentration), proteoglycan content (measured as hexosamine concentration), and proteoglycan synthesis (measured as [35S]sulfate incorporation) varied according to the site sampled. Cartilage from the transverse ridge of the head of the third metacarpal bone and the radial facet of the third carpal bone had the lowest hexosamine concentration, whereas rate of proteoglycan synthesis was lowest in cartilage from the transverse ridge of the head of the third metacarpal bone and the distal articular surface of the radial carpal bone. Repair tissue filling a full-thickness cartilage defect at 6 weeks was highly cellular. It was low in proteoglycan content, but was actively synthesizing these macromolecules. In contrast, the cartilage surrounding a partial-thickness defect was unchanged 6 weeks after the original defect was made.

Evaluation of pudendal reflexes and effects of pudendal branch conditioning on those reflexes was carried out in 2 studies. In the first study of pudendal reflexes, 20 adult male and female mixed-breed cats underwent surgical isolation of the anal branch, urethral branch, and distal trunk (consisting primarily of the dorsal nerve of the penis/clitoris) of the pudendal nerve. Reflexes were tested in all possible ipsilateral and contralateral test-response combinations. Latency values and effects of increasing stimulus rate on response amplitude were recorded. Reflexes were detected in all combinations, with response latencies between 6.3 and 13.0 ms. Response amplitudes were diminished at stimulus rates of 3 to 5 Hz, and responses were apparently abolished at 4 to 16 Hz, suggesting that pudendal reflexes are polysynaptic. In the second study of conditioning effects, 9 adult male and female mixed-breed cats underwent preparation similar to that for study 1. A train of conditioning stimuli was applied to branches of the pudendal nerve prior to attempting to induce reflex responses, as performed in study 1. Conditioning completely abolished reflex responses for a period of 70 to 130 ms. Reflex responses were diminished in amplitude, compared with those observed during preconditioning trials, for 180 to 300 ms after conditioning.

The use of periosteal autografts to resurface osteochondral defects was investigated in 10 horses (2 to 3 years old), and the repair tissue was characterized morphologically. Middle carpal joint arthrotomies were made, and osteochondral defects were induced bilaterally on the distal articular surface of each radial carpal bone. Each defect measured approximatively 1 cm2 and extended 3 mm into the subchondral bone plate. Residual subchondral bone plate of control and principal defects was perforated by drilling. A sterile fibrin adhesive was made by mixing a fibrinogen component and a thrombin component. A periosteal autograft was harvested from the proximal portion of the tibia and was glued onto the recipient osseous surface, with its cambium facing the joint cavity. Control defects were glued, but not grafted. Horses were walked 1 hour daily on a walker, starting at postoperative week 7 and continuing for 9 weeks. Sixteen weeks after the grafting procedure was done, carpal radiography was performed, after which horses were euthanatized. Quality of repair tissue of control and grafted defects was evaluated and compared grossly, histologically, and histochemically. Using a reticule, the proportions of various repair tissue types filling each defect were quantitated. Seven weeks after the grafting procedure was done, bilateral arthroscopy revealed synovial adhesions and marginal pannus formation in control and grafted defects. None of the autograft was found floating unattached within the respective middle carpal joints. At 16 weeks, the gross appearance of most grafted and nongrafted defects was similar, and repair was dominated by a fibrous pannus. In 4 grafted defects, bone had formed either concentrically within the defect or eccentrically in the fibrous adhesions between the defect and the joint margin. Histologically, all grafted and nongrafted defects were repaired similarly by infiltration of a mixture of fibrous tissue, fibrocartilage, and bone. Fibrous tissue was the predominant tissue in most defects and its mean proportion was 56 and 59% in the grafted and nongrafted defects, respectively. Fibrocartilaginous tissue in the deeper layers approximated 20%, and woven bone at the base of the defect was 20% in all defects. Histochemically, difference in staining for proteoglycans was not observed between grafted and nongrafted defects. Little remaining original periosteal graft tissue was evident at the defect sites. The only distinguishing feature of grafted defects was the presence of islands of bone formation either at the defect site (n = 2 horses), or in somewhat dorsally displaced tissue that was incorporated in fibrous adhesions (n = 2 horses). It was concluded that use of periosteal autograft did not improve the healing of osteochondral defects of the distal portion of the radial carpal bone. The repair tissue produced in grafted and nongrafted defects was similar and was principally fibrous in nature.

The effects of method of seminal collection and a diuretic on retrograde flow of spermatozoa into the urinary bladder of rams were examined. In experiment 1, semen and urine were collected from 8 rams during the nonbreeding season. Prior to seminal collection, all rams were given furosemide and a sample of urine was obtained during micturition. Semen was then collected from each ram with an artificial vagina or by electroejaculation in alternate weeks for 4 weeks, and the urine released during the first postseminal collection micturition was collected in 4 consecutive samples. The volume of electroejaculates was larger (P < 0.0001) than the volume of ejaculates, but the total number of spermatozoa in the electroejaculate or in the ejaculate were not different (P > 0.1). Urine obtained before seminal collection was azoospermic or contained few, nonmotile spermatozoa (mean +/- SD = 0.053 +/- 0.114 x 10(6)/ml). The adjusted spermatozoal concentration (mean +/- SD = 1.630 +/- 2.258 X 10(6)/ml) in the urine collected after seminal collection was 31 times higher (P < 0.0001) and there were motile spermatozoa in most (97%) of the samples. The spermatozoal concentration in sequential samples of urine was not different (P > 0.1) between samples and was not affected (P > 0.1) by the method of seminal collection. There was a trend, approaching significance (P = 0.052), for an effect of method of seminal collection on the percentage of retrograde flow. Retrograde flow ranged from 0.21 to 19.38% when semen was collected with an artificial vagina and from 0.03 to 94.60% when semen was collected by electroejaculation and varied (P = 0.02) among rams within the 2 methods of seminal collection. In experiment 2, the 8 rams used in experiment 1 were given injections of 0.9% physiologic saline solution or furosemide in alternate weeks prior to seminal collection with an artificial vagina. Furosemide increased (P = 0.009) the volume of urine voided during the first postejaculation micturition, but did not influence (P > 0.1) the time from exposure of rams to the teaser to ejaculation, seminal characteristics, number of spermatozoa in the urine, or the percentage of retrograde flow. There was a trend (P < 0.1) for more rams to have motile spermatozoa in the postejaculation urine after treatment with furosemide. Administration of furosemide prior to seminal collection facilitates the noninvasive collection of pre- and postejaculation samples of urine for the determination of retrograde flow.

Polymerase chain reaction was used to detect an economically important herpesvirus, channel catfish virus (CCV). A segment of the viral DNA was sequenced and oligonucleotide primers were produced from that sequence. After the primers were tested for the possibility of hybridization to catfish DNA, they were used to prime the polymerase chain reaction, using pure CCV DNA, CCV DNA added to catfish DNA, and DNA from catfish infected and not infected with CCV. In all cases, the method proved to be simple and sensitive in its detection of CCV DNA. When catfish DNA was present, < 0.1 pg of CCV DNA was detectable. Channel catfish virus DNA in a latent carrier of CCV was readily detectable.

Enzyme-linked immunosorbent assay screening of antibody produced against aflatoxin was accomplished by a new and simple procedure. To demonstrate the new indirect ELISA technique used, antibody against aflatoxin M1 was produced in female BALB/CJ mice by immunization with an aflatoxin M1-bovine serum albumin conjugate. Instead of coating test-plate wells with purified antibody (direct ELISA) or synthesizing a second protein-aflatoxin conjugate (aflatoxin M1-poly-L-lysine) to coat test-plate wells, wells were coated with the readily available aflatoxin M1-bovine serum albumin and aflatoxin B1-bovine vine serum albumin. This method, applicable for any aflatoxin conjugated by the common cyclopentano-carboxymethoxyl-oxime technique, eliminates the more time-consuming and technically difficult portions of earlier direct and indirect ELISA. The new technique can be valuable in continued efforts toward development of new and improved immunoassays against aflatoxin metabolites.

Sixty-three drugs, belonging to 10 chemical classes, were tested in vitro to determine effects on phagocytosis of 32P-labeled Staphylococcus aureus by neutrophils isolated from milk. Within each class, the number of antibiotics tested were: nonsteroidal anti-inflammatory drugs (NSAID; 8), peptolids (2), aminoglycosides (8), tetracyclines and fusidic acid (4), beta-lactam antibiotics (25), secretolytic agents (2), macrolides (5), polypeptides (2), and antibacterial quinolones (8). Percentage of phagocytosis was determined after incubating (2 hours at 37 C) 12.5 X 10(6) viable neutrophils, 200 X 10(6) 32P-labeled S aureus with antibiotics and 5% skimmed milk. Concentrations of antibiotics tested were 1,000, 500, and 10 microgram/ml of incubation media. When compared with nonantibiotic controls at the highest drug concentration, the NSAID acetylsalicylic acid and centrophenoxine increased phagocytosis 23.2 and 8.8%, respectively, and benzydamine, indomethacin, phenylbutazone, ibuprofen, and acetominophen decreased phagocytosis 22.8, 14.2, 9.8, 27.0, and 18.2%, respectively. The peptolids novobiocin and pristinamycin decreased phagocytosis 24.5 and 22.0%, respectively. The aminoglycosides tobramycin, amikacin, and gentamicin decreased phagocytosis 21.1, 15.4, and 19.2%, respectively. For the tetracyclines and fusidic acid, minocycline and doxycycline decreased phagocytosis 39.8 and 54.2%, respectively. The beta-lactam antibiotics carfecillin, cephapirin sodium, and cephacetrile sodium decreased phagocytosis 11.2, 12.8, and 23.8%, respectively. The secretolytic agent, bromhexin, increased phagocytosis 10.8%. These data indicate that the potential for enhanced phagocytosis exists through use of some NSAID, and for depressed phagocytosis through use of aminoglycosides, peptolids, tetracyclines, and beta-lactams, as well as certain other NSAID.

We developed a single-pedicle, axial pattern tubed skin flap that could be transferred to an in vitro perfusion apparatus. On the basis of results of prosections, angiography, contact radiography, and surviving-length studies, it was concluded that a single-pedicle, axial pattern skin flap measuring 4 cm X 12 cm incorporating the caudal superficial epigastric artery would survive to its entire length. Subsequently, a surgical (stage 1) procedure was developed for the routine preparation of single-pedicle, axial pattern tubed skin flaps. Healing after the stage-1 procedure was evaluated by visual inspection and fluorescein angiography. Stage-1 procedures were performed successfully 149 of 160 (93%) times. A second surgical (stage 2) procedure was developed for routine cannulation of the caudal superficial epigastric artery and harvest of the tubed skin flap. Stage-2 procedures were performed successfully 136 of 144 (94%) times.

Thirty-six of 50 young equids examined at necropsy for gross pathologic and histopathologic evidence of osteochondrosis were determined to have lesions characteristic of this disorder in the distal joints of the tarsus. Abnormalities ranged from retained endochondral cores underlying undisturbed articular cartilage surfaces to clefts, subchondral osseous cyst-like lesions, and cartilage ulceration. Our findings supported the conclusion that osteochondrosis may cause spavin in the juvenile equid.

The steady-state response characteristics of a pulse oximeter were evaluated on intestinal segments of seven clinically normal halothane-anesthetized horses. Arterial oxygen tension > 200 mm of Hg, end tidal carbon dioxide from 30 to 35 mm of Hg, and systemic mean arterial pressure > 70 mm of Hg were maintained throughout the recording periods. Values for percentage of pulse oximeter oxygen saturation, pulsatile blood flow, and percentage of signal strength were recorded from jejunum, ileum, cecum, left ventral colon, left dorsal colon, and descending colon. Probe placement on intestinal segments was recorded as over or not over visible subserosal or transmural vessels. There was no significant difference between median values on the basis of vessel codes for pulse oximeter oxygen saturations, pulsatile flow, and signal strength. Median values recorded for pulse oximeter oxygen saturation were 93% from jejunum and ileum and 95% from cecum, left ventral colon, left dorsal colon, and descending colon; median values for pulsatile flow were 576 from jejunum, 560 from ileum, 560 from cecum, 574 from left ventral colon, 578 from left dorsal colon, and 560 from descending colon; median values for signal strength were 50% from jejunum, 67.5% from ileum, 60% from cecum, 75% from left ventral colon, 50% from left dorsal colon, and 52.5% from descending colon. Median values obtained from each anatomic location were not significantly different for pulsatile flow or signal strength. Median pulse oximetry oxygen values recorded from jejunum and ileum were significantly lower than values obtained from other intestinal segments. When calculated arterial oxygen saturation was compared with oxygen saturation determined by the pulse oximeter, pulse oximeter oxygen saturation was consistently lower by 6.7% (jejunum and ileum) and 4.7% (cecum, left ventral colon, left dorsal colon, and descending colon). Equine and human absorption spectra were generated and compared for reduced hemoglobin and oxyhemoglobin at wavelengths of 600 nm (red) to 950 nm (infrared). Extinction coefficients calculated at wavelengths used by the pulse oximeter (660 nm and 940 nm) were nearly identical. The pulse oximeter is a self-calibrating instrument that displays oxygen saturation, heart rate, plethysmographic waveform, and signal strength indicator. Probe application was rapid and easy. Response time for the appearance of a plethysmographic waveform ranged from 5 to 25 seconds.