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Topic:Physiology

The physiology of horses encompasses the study of the biological functions and processes that occur within the equine body. This includes the examination of various systems such as the cardiovascular, respiratory, musculoskeletal, digestive, and nervous systems. Understanding equine physiology is essential for comprehending how horses adapt to different environmental conditions, perform physical activities, and respond to health challenges. Research in this field often focuses on the mechanisms of energy metabolism, thermoregulation, and muscle function during exercise, as well as the physiological responses to stress and disease. This page compiles peer-reviewed research studies and scholarly articles that explore the diverse aspects of equine physiology, providing insights into the biological processes that support the health and performance of horses.
Correlation of racing performance with radiographic changes in the proximal sesamoid bones of 487 Thoroughbred yearlings.
Equine veterinary journal    July 26, 2003   Volume 35, Issue 4 350-353 doi: 10.2746/042516403776014262
Spike-Pierce DL, Bramlage LR.Proximal sesamoiditis has been a common and confusing radiographic diagnosis in Thoroughbred yearling surveys. This study categorised the radiographic appearance of yearling proximal sesamoid bones and determined their effect on the number of races started and earning potential for racing at ages 2 and 3 years. Objective: Yearlings with moderate to severe proximal sesamoiditis have decreased performance at age 2 and 3 years. Methods: Survey radiographs of 487 Thoroughbred yearlings were reviewed. Proximal sesamoid bones were examined and findings placed into 7 categories, 5 of which measured s...
Blood/gas partition coefficients of halothane, isoflurane and sevoflurane in horse blood.
British journal of anaesthesia    July 25, 2003   Volume 91, Issue 2 276-278 doi: 10.1093/bja/aeg151
Bergadano A, Lauber R, Zbinden A, Schatzmann U, Moens Y.Blood/gas partition coefficients (lambda(b/g)) for volatile agents in horse blood are reported for halothane but not for isoflurane and sevoflurane. We measured the lambda(b/g) of halothane, isoflurane and sevoflurane in the blood of fasted horses. The correlation with age, weight and some haematological and biochemical variables was studied. The temperature correction factor for isoflurane solubility was calculated. Methods: Twenty-four horses were randomly allocated to halothane (n=8), isoflurane (n=8) or sevoflurane (n=8). Blood samples were taken after 10 h' fasting. Calculation of lambda(...
Equine articular cartilage chondrocytes: opening the black box.
Equine veterinary journal    July 24, 2003   Volume 35, Issue 5 425-428 doi: 10.2746/042516403775600442
Hall AC, Bush PG, Davidson ME, Kempson SA.No abstract available
Effects of lipopolysaccharide and phenylbutazone on gastric contents in the horse.
Equine veterinary journal    July 24, 2003   Volume 35, Issue 5 472-475 doi: 10.2746/042516403775600488
Doherty TJ, Andrews FM, Blackford JT, Rohrbach BW, Sandin A, Saxton AM.Endotoxaemia causes a disruption of gastrointestinal motility in the horse but there is no information on its effects on gastric secretion. Lipopolysaccharide (LPS) administration is known to affect gastric secretion in other species. Objective: That LPS, a toxic component of Gram-negative bacteria, would reduce gastric acid secretion and that pretreatment with phenylbutazone (PBZ) would block the effects of LPS. Methods: The effects of LPS and PBZ on gastric contents were investigated in fasted, mature horses, with permanent gastric cannulae. Horses were pretreated with either saline or PBZ 1...
The effects of maternal age and parity on placental and fetal development in the mare.
Equine veterinary journal    July 24, 2003   Volume 35, Issue 5 476-483 doi: 10.2746/042516403775600550
Wilsher S, Allen WR.The normality of equine placentation is essential for fetal health and development. Substantial information exists on the gross morphological status of the placenta but few studies have addressed the problem of degenerative lesions that interfere with placental morphology and placental efficiency. Objective: Degenerative changes in the endometrium with increasing age and parity are reflected in the morphology and density of the placental microcotyledons. Objective: To assess placental efficiency on the basis of foal birthweight as a function of total microscopic area of fetomaternal contact. M...
Homeostasis of intracellular Ca2+ in equine chondrocytes: response to hypotonic shock.
Equine veterinary journal    July 24, 2003   Volume 35, Issue 5 439-443 doi: 10.2746/042516403775600541
Wilkins RJ, Fairfax TP, Davies ME, Muzyamba MC, Gibson JS.Ca2+ homeostasis in articular chondrocytes affects synthesis and degradation of the cartilage matrix, as well as other cellular functions, thereby contributing to joint integrity. Although it will be affected by mechanical loading, the sensitivity of intracellular Ca2+ concentration ([Ca2+]i) in equine articular chondrocytes to many stimuli remains unknown. Objective: An improved understanding of Ca2+ homeostasis in equine articular chondrocytes, and how it is altered during joint loading and pathology, will be important in understanding how joints respond to mechanical loads. Methods: [Ca2+]i...
Dynamic compressive strain inhibits nitric oxide synthesis by equine chondrocytes isolated from different areas of the cartilage surface.
Equine veterinary journal    July 24, 2003   Volume 35, Issue 5 451-456 doi: 10.2746/042516403775600532
Wiseman M, Henson F, Lee DA, Bader DL.Chondrocytes within articular cartilage respond to the mechanical stresses associated with normal joint loading via a series of signalling pathways. Specific biomolecules, such as nitric oxide (NO), have been implicated in these mechanotransduction processes. It has been shown that the synthesis of NO can be inhibited by dynamic compressive strain of chondrocytes in vitro which, in turn, leads to an up-regulation of specific metabolic parameters. Objective: Chondrocytes isolated from different joint locations and seeded in agarose constructs respond in a distinct manner to the application of d...
Spontaneous repair of the atrophic contralateral ovary without ovariectomy in the case of a granulosa theca cell tumor (GTCT) affected mare.
The Journal of veterinary medical science    July 18, 2003   Volume 65, Issue 6 749-751 doi: 10.1292/jvms.65.749
Hoque S, Derar RI, Osawa T, Taya K, Watanabe G, Miyake Y.A 21 year old thoroughbred mare with granulosa theca cell tumor (GTCT) in the right side and atrophic contralateral ovary was investigated in this study. After arrival at our laboratory on 10th December 1999, the clinical diagnosis of GTCT was examined by rectal palpation and ultrasonographic image of ovaries. Plasma from peripheral blood was collected in the breeding and non-breeding seasons for hormonal analysis. The results showed that the contralateral ovary regained normal activity without any treatment of the GTCT affected ovary and contained follicles showing different sizes 19 months l...
Effects of hypothyroidism and withholding of feed on plasma lipid concentrations, concentration and composition of very-low-density lipoprotein, and plasma lipase activity in horses.
American journal of veterinary research    July 15, 2003   Volume 64, Issue 7 823-828 doi: 10.2460/ajvr.2003.64.823
Frank N, Sojka JE, Latour MA.To evaluate selected concentrations of blood lipids and lipase activities in euthyroid and hypothyroid horses deprived of feed for 96 hours. Methods: 4 healthy adult mares and 4 thyroidectomized adult mares. Methods: Horses were deprived of feed for 96 hours. Blood samples were collected at 24-hour intervals and analyzed to determine concentrations of non-esterified fatty acid (NEFA), triglyceride (TG), total cholesterol (TC), and very-low-density lipoprotein (VLDL) as well as composition of VLDL. Plasma lipase activities were measured after feed was withheld for 96 hours and 12 days after res...
Assessment of three variations of the 1,9-dimethylmethylene blue assay for measurement of sulfated glycosaminoglycan concentrations in equine synovial fluid.
American journal of veterinary research    July 15, 2003   Volume 64, Issue 7 900-906 doi: 10.2460/ajvr.2003.64.900
Oke SL, Hurtig MB, Keates RA, Wright JR, Lumsden JH.To determine whether 3 variations of the 1,9-dimethylmethylene blue (DMMB) assay yield comparable results when measuring sulfated glycosaminoglycan (sGAG) concentrations in equine synovial fluid (SF). Methods: 25 samples of SF collected from affected joints of 13 horses and 13 samples of SF collected from nonaffected (control) joints of 4 horses. Methods: Sulfated glycosaminoglycan concentrations were measured by the direct spectrophotometric (ie, Farndale), microplate, and indirect DMMB assays in samples of SF collected from normal and affected joints and in samples digested with nucleases, p...
Submural histopathologic changes attributable to peracute laminitis in horses.
American journal of veterinary research    July 15, 2003   Volume 64, Issue 7 829-834 doi: 10.2460/ajvr.2003.64.829
Morgan SJ, Hood DM, Wagner IP, Postl SP.To describe submural histopathologic changes attributable to peracute laminitis in horses. Methods: 20 adult horses. Methods: A concurrent-control design was used to compare laminar lesions in 10 horses subjected to carbohydrate-induced laminitis with laminar characteristics of 10 sex- and aged-matched control horses with normal feet. Horses in the treatment group were administered an overload of carbohydrate. Tissues were obtained by biopsy 4 to 8 hours after onset of lameness or 72 hours after administration of the carbohydrate overload when lameness did not develop. Sections were stained wi...
Development of a solid-phase assay for measurement of sulfated glycosaminoglycan concentrations in equine synovial fluid.
American journal of veterinary research    July 15, 2003   Volume 64, Issue 7 894-899 doi: 10.2460/ajvr.2003.64.894
Oke SL, Hurtig MB, Keates RA, Wright JR.To develop a new 1,9-dimethylmethylene blue (DMMB) assay for measurement of sulfated glycosaminoglycan (sGAG) concentrations in equine synovial fluid (SF) by use of membrane technology and to compare the assay's ability to measure sGAG concentrations with that of 2 other established DMMB assays. Methods: 25 samples of SF collected from affected joints of 14 horses and 13 samples of SF collected from nonaffected (control) joints of 4 horses. Methods: A solid-phase DMMB assay was developed to measure sGAG concentrations in SE Results for the assay were then compared with results obtained by use ...
Influence of topically applied cold treatment on core temperature and cell viability in equine superficial digital flexor tendons.
American journal of veterinary research    July 15, 2003   Volume 64, Issue 7 835-844 doi: 10.2460/ajvr.2003.64.835
Petrov R, MacDonald MH, Tesch AM, Van Hoogmoed LM.To determine rate and degree of cooling for the superficial digital flexor tendon (SDFT) during a standard cryotherapy application in horses and evaluate in vitro effects of cooling on survival of tendon cells. Methods: 6 limbs of 5 adult horses and cultured cells obtained from SDFT of 3 adult horses during necropsy. Methods: In vivo data were acquired by use of a thermocouple temperature probe inserted into the SDFT of a forelimb of each standing sedated horse. After baseline temperatures were recorded, a commercial compression splint with circulating coolant was placed on each selected limb,...
Glucose clearance in grazing mares is affected by diet, pregnancy, and lactation.
Journal of animal science    July 12, 2003   Volume 81, Issue 7 1764-1771 doi: 10.2527/2003.8171764x
Hoffman RM, Kronfeld DS, Cooper WL, Harris PA.The glucose tolerance test in the horse may be used to determine metabolic responses to diet, disease, or physiologic state. The objective of this study was to determine the effect of reproductive stage (gestation and lactation) and supplemental dietary energy source (sugar and starch [SS] or fiber and fat [FF]) on glucose metabolism in grazing mares using an oral glucose tolerance test. Twelve mares, six on each supplement, were examined on three occasions: one in the third trimester of pregnancy, the second in early lactation, and the third in late lactation. During each test, venous samples...
Pregnancy outcome in mares following insemination deep in the uterine horn with low numbers of sperm selected by glass wool/Sephadex filtration, Percoll separation or absolute number.
Animal reproduction science    July 11, 2003   Volume 79, Issue 1-2 103-109 doi: 10.1016/s0378-4320(03)00086-1
Nie GJ, Johnson KE, Wenzel JG.Mares were inseminated deep in the uterine horn with 25 million sperm selected by glass wool/Sephadex (GWS) filtration, Percoll separation (PS) or absolute number (AN). Deep-horn insemination using a low-volume, smooth tipped, flexible pipette/catheter delivery system allowed more efficient use of stallion sperm and reduced post-breeding uterine reaction in mares. Mares were pregnant in 15/30, 13/30 and 10/30 cycles for GWS, PS and AN selection methods, respectively. Sperm selection method did not effect pregnancy outcome (P=0.422). However, sperm selected for deep-horn insemination by filtrat...
Amyloid protofilaments from the calcium-binding protein equine lysozyme: formation of ring and linear structures depends on pH and metal ion concentration.
Journal of molecular biology    July 10, 2003   Volume 330, Issue 4 879-890 doi: 10.1016/s0022-2836(03)00551-5
Malisauskas M, Zamotin V, Jass J, Noppe W, Dobson CM, Morozova-Roche LA.The calcium-binding equine lysozyme has been found to undergo conversion into amyloid fibrils during incubation in solution at acidic pH. At pH 4.5 and 57 degrees C, where equine lysozyme forms a partially unfolded molten globule state, the protein forms protofilaments with a width of ca. 2 nm. In the absence of Ca(2+) the protofilaments are present as annular structures with a diameter of 40-50 nm. In the presence of 10 mM CaCl(2) the protofilaments of equine lysozyme are straight or curved; they can assemble into thicker threads, but they do not appear to undergo circularisation. At pH 2.0, ...
Initial orthostatic hypotension in a 37-year old horse rider.
Clinical autonomic research : official journal of the Clinical Autonomic Research Society    July 5, 2003   Volume 12, Issue 5 404 doi: 10.1007/s10286-002-0062-6
Krediet CT.No abstract available
Regional antibody and cellular immune responses to equine influenza virus infection, and particle mediated DNA vaccination.
Veterinary immunology and immunopathology    July 5, 2003   Volume 94, Issue 1-2 47-62 doi: 10.1016/s0165-2427(03)00060-6
Soboll G, Horohov DW, Aldridge BM, Olsen CW, McGregor MW, Drape RJ, Macklin MD, Swain WF, Lunn DP.We have previously demonstrated that hemagglutinin (HA) gene vaccination and influenza virus infection generate protective antibody responses in equids. However, these antibody responses differ substantially in that particle mediated DNA vaccination does not induce an immunoglobulin A (IgA) response. A study was performed to investigate the regional immunoregulatory mechanisms associated with these different immune responses. Ponies were either vaccinated with equine HA DNA vaccines at skin and mucosal sites, infected with influenza virus or left untreated and influenza-specific antibody respo...
Day of cycle affects changes in equine intrauterine pressure in response to teasing.
Theriogenology    July 2, 2003   Volume 60, Issue 4 727-733 doi: 10.1016/s0093-691x(03)00076-1
Stecco R, Paccamonti D, Gutjahr S, Pinto CR, Eilts B.Oxytocin is released in response to teasing during both estrus and diestrus in mares, and at least during estrus, teasing results in an increase in electromyographic activity in the uterus. Exogenous oxytocin causes an increase in intrauterine pressure and prior studies have shown that this response is correlated to the day of the estrous cycle. To determine if teasing causes an increase in intrauterine pressure and if this response varies by day of the cycle, intrauterine pressure was measured while mares were teased with a stallion 2 days before ovulation, on the day ovulation was detected a...
Effect of varying initial drink volume on rehydration of horses.
Physiology & behavior    July 2, 2003   Volume 79, Issue 2 135-142 doi: 10.1016/s0031-9384(03)00085-4
Butudom P, Axiak SM, Nielsen BD, Eberhart SW, Schott HC.Body mass (BM), water intake (WI), and plasma osmolality (P(osm)) and electrolyte concentrations were measured in six 2-year-old Arabian horses provided either 4 l, 8 l, or an unlimited amount of water (UW) for drinking during the initial 5 min of recovery from 45-km of treadmill exercise. After weighing, horses were placed in a stall and further WI between 20 and 60 min of recovery was measured. During exercise, horses lost 3.3+/-0.3%, 3.2+/-0.1%, and 3.3+/-0.2% (P>.05) of BM and P(osm) increased by 7.2+/-0.5, 7.9+/-0.8, and 7.7+/-0.5 mOsm/kg (P>.05) for 4 l, 8 l, and UW, respectively. ...
Transrectal Doppler sonography of uterine blood flow during early pregnancy in mares.
Theriogenology    July 2, 2003   Volume 60, Issue 4 597-605 doi: 10.1016/s0093-691x(03)00080-3
Bollwein H, Mayer R, Stolla R.Transrectal color Doppler sonography was used for the noninvasive investigation of uterine blood flow in five mares. Both the left and right uterine arteries were scanned to obtain blood flow velocity waveforms during two consecutive estrous cycles and two early pregnancies in each mare. Blood flow was expressed as the time-averaged maximum velocity (TAMV) and the resistance index (RI). In all pregnancies the embryonic vesicle could be detected for the first time on Day 11 (day of ovulation: Day 0). No differences in mean TAMV and RI values of both uterine arteries were observed in comparison ...
Clostridium perfringens alpha-toxin-induced hemolysis of horse erythrocytes is dependent on Ca2+ uptake.
Biochimica et biophysica acta    July 2, 2003   Volume 1613, Issue 1-2 79-86 doi: 10.1016/s0005-2736(03)00140-8
Ochi S, Oda M, Nagahama M, Sakurai J.Clostridium perfringens alpha-toxin is able to lyse various erythrocytes. Exposure of horse erythrocytes to alpha-toxin simultaneously induced hot-cold hemolysis and stimulated production of diacylglycerol and phosphorylcholine. When A23187-treated erythrocytes were treated with the toxin, these events were dependent on the concentration of extracellular Ca2+ . Incubation with the toxin of BAPTA-AM-treated horse erythrocytes caused no hemolysis or production of phosphorylcholine, but that of the BAPTA-treated erythrocytes did. When Quin 2-AM-treated erythrocytes were incubated with the toxin i...
Assessment of corneal thickness, intraocular pressure, optical corneal diameter, and axial globe dimensions in Miniature Horses.
American journal of veterinary research    June 28, 2003   Volume 64, Issue 6 661-665 doi: 10.2460/ajvr.2003.64.661
Plummer CE, Ramsey DT, Hauptman JG.To determine corneal thickness, intraocular pressure (IOP), and horizontal and vertical corneal diameter (HCD and VCD) and to obtain axial measurements of the anterior chamber depth (ACD), crystalline lens thickness (CLT), vitreous chamber depth (VtCD), and axial globe length (AGL) in eyes of Miniature Horses. Methods: 41 healthy Miniature Horses. Methods: Ocular component measurements were obtained via ultrasonic pachymetry, applanation tonometry, ultrasound, and by use of a Jameson caliper. Results: Mean IOP and corneal thickness for all eyes were 26.0 mm Hg and 785.6 microm, respectively. T...
Evaluation of beta3-adrenoceptor-mediated relaxation in intact and endotoxin-treated equine digital veins.
American journal of veterinary research    June 28, 2003   Volume 64, Issue 6 708-714 doi: 10.2460/ajvr.2003.64.708
Mallem MY, Gogny M, Gautier F, Bucas V, Desfontis JC.To investigate the functional expression of beta3-adrenoceptors (beta3-ARs) in equine digital veins (EDVs) and to examine whether beta3-AR relaxation was altered in EDVs incubated with endotoxin. Methods: Forelimbs obtained from 30 horses. Methods: Forelimbs were obtained from horses in an abattoir. Equine digital veins were carefully removed from distal portions of the forelimbs. Rings of dissected EDVs were mounted in 5-mL organ baths to record isometric tension in the presence of various beta3-AR agonists (SR 58611A, ZD 2079, and ZM 215001). Results: In intact EDVs, isoprenaline, SR 58611A,...
Comparison of nociceptive withdrawal reflexes and recruitment curves between the forelimbs and hind limbs in conscious horses.
American journal of veterinary research    June 28, 2003   Volume 64, Issue 6 700-707 doi: 10.2460/ajvr.2003.64.700
Spadavecchia C, Arendt-Nielsen L, Andersen OK, Spadavecchia L, Doherr M, Schatzmann U.To compare nociceptive withdrawal reflexes (NWRs) evoked from the distal aspect of the left forelimb and hind limb in conscious standing horses and to investigate NWR recruitment for graded electrical stimulation intensities. Methods: 20 adult horses. Methods: Surface electromyographic (EMG) activity evoked by transcutaneous electrical stimulation of the digital palmar (or plantar) nerve was recorded from the common digital extensor and cranial tibial muscles. Stimuli consisted of 25-millisecond train-of-5 constant current pulses. Current intensity was gradually increased until NWR threshold i...
Influence of glucosamine on matrix metalloproteinase expression and activity in lipopolysaccharide-stimulated equine chondrocytes.
American journal of veterinary research    June 28, 2003   Volume 64, Issue 6 666-671 doi: 10.2460/ajvr.2003.64.666
Byron CR, Orth MW, Venta PJ, Lloyd JW, Caron JP.To characterize potential mechanisms of action of glucosamine inhibition of matrix metalloproteinase (MMP) expression and activity in lipopolysaccharide (LPS)-stimulated equine chondrocytes. Methods: Chondrocytes cultured from samples of metacarpophalangeal articular cartilage collected from cadaveric limbs of horses. Methods: The effect of glucosamine on MMP activity in conditioned medium from LPS-stimulated cartilage explants was determined by a colorimetric assay with azocoll substrate. Treatments consisted of negative and positive controls, glucose (50 mM), and glucosamine (50, 25, 6.25, 3...
Reactive oxygen species and cryopreservation promote DNA fragmentation in equine spermatozoa.
Journal of andrology    June 27, 2003   Volume 24, Issue 4 621-628 doi: 10.1002/j.1939-4640.2003.tb02714.x
Baumber J, Ball BA, Linfor JJ, Meyers SA.The objective of this study was to examine the effect of reactive oxygen species (ROS) and cryopreservation on DNA fragmentation of equine spermatozoa. In experiment 1, equine spermatozoa were incubated (1 hour, 38 degrees C) according to the following treatments: 1) sperm alone; 2) sperm + xanthine (X, 0.3 mM)-xanthine oxidase (XO, 0.025 U/mL); 3) sperm + X (0.6 mM)-XO (0.05 U/mL); and 4) sperm + X (1 mM)-XO (0.1 U/mL). In experiment 2, spermatozoa were incubated (1 hour, 38 degrees C) with X (1 mM)-XO (0.1 U/mL) and either catalase (200 U/mL), superoxide dismutase (SOD, 200 U/mL), or reduced...
3-methylindole induces transient olfactory mucosal injury in ponies.
Veterinary pathology    June 26, 2003   Volume 40, Issue 4 363-370 doi: 10.1354/vp.40-4-363
Miller MA, Kottler SJ, Ramos-Vara JA, Johnson PJ, Ganjam VK, Evans TJ.Response to 3-methylindole (3MI) varies among species. Mice recover from 3MI-induced bronchiolar epithelial injury but sustain persistent olfactory mucosal injury with scarring and epithelial metaplasia. In contrast, 3MI induces obliterative bronchiolitis in horses and ponies, but olfactory mucosal injury has not been reported. To evaluate the effect of 3MI on equine olfactory mucosa, ponies were dosed orally with 100 mg 3MI/kg (n = 9) or corn oil vehicle (n = 6). All ponies treated with 3MI developed obliterative bronchiolitis with mild olfactory injury. By 3 days after 3MI dosing, olfactory ...
MultiPipMaker and supporting tools: Alignments and analysis of multiple genomic DNA sequences.
Nucleic acids research    June 26, 2003   Volume 31, Issue 13 3518-3524 doi: 10.1093/nar/gkg579
Schwartz S, Elnitski L, Li M, Weirauch M, Riemer C, Smit A, Green ED, Hardison RC, Miller W.Analysis of multiple sequence alignments can generate important, testable hypotheses about the phylogenetic history and cellular function of genomic sequences. We describe the MultiPipMaker server, which aligns multiple, long genomic DNA sequences quickly and with good sensitivity (available at http://bio.cse.psu.edu/ since May 2001). Alignments are computed between a contiguous reference sequence and one or more secondary sequences, which can be finished or draft sequence. The outputs include a stacked set of percent identity plots, called a MultiPip, comparing the reference sequence with sub...
Histochemical detection of glycoconjugates in the male reproductive system of the horse.
Journal of veterinary science    June 24, 2003   Volume 4, Issue 1 21-28 
Ha TY, Ahn MJ, Lee YD, Yang JH, Kim HS, Shin TK.Lectins are glycoproteins of plant and animal origin that have the ability to bind specific carbohydrate residues of cell glycoconjugates, particularly in terminal positions. In this study, the binding of lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I), was studied in the reproductive systems of male thoroughbred horses.DBA was detected in the stereocilia of the caput and corpus epididymis, and in the vas deferens. It was weakly detected in connective...