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Topic:In Vivo

In vivo studies involving horses refer to research conducted within living organisms to understand various biological processes, disease mechanisms, and therapeutic interventions in equine species. These studies provide insights into the physiological and pathological responses of horses under natural conditions. In vivo research encompasses a range of investigations, including pharmacokinetics, disease modeling, and the evaluation of treatment efficacy. It often involves the use of controlled environments to simulate real-life scenarios while observing the effects of specific variables on equine health. This page gathers peer-reviewed research articles and scholarly studies that explore the methodologies, findings, and implications of in vivo research in horses, contributing to the advancement of equine science and veterinary medicine.
Immunoepidemiology of the equine tapeworm Anoplocephala perfoliata: age-intensity profile and age-dependency of antibody subtype responses.
Parasitology    January 1, 1997   Volume 114 ( Pt 1) 89-94 doi: 10.1017/s0031182096008086
Proudman CJ, Holmes MA, Sheoran AS, Edwards SE, Trees AJ.The equine intestinal cestode Anoplocephala perfoliata has been the subject of recent epidemiological and immunological studies because of its suspected association with intestinal disease in the horse. We have previously shown that the IgG(T) subtype antibody response to the 12/13 kDa component of the parasite excretory/secretory (E/S) antigen is positively correlated with parasite intensity. In this study, we utilize that correlation to examine the changes in natural infection intensity with age. Infection intensity based on IgG(T) responses showed a triphasic age-dependency pattern with pea...
Effect of treatment with erythromycin on bronchoalveolar lavage fluid cell populations in foals.
American journal of veterinary research    January 1, 1997   Volume 58, Issue 1 56-61 
Lakritz J, Wilson WD, Watson JL, Hyde DM, Mihalyi J, Plopper CG.To determine whether oral administration of erythromycin alters the inflammatory response to bronchoalveolar lavage (BAL) in young horses. Methods: 12 healthy, unweaned, mixed-breed foals of either sex, between 2 and 4 months old. Methods: BAL was performed; 250 ml of phosphate-buffered saline solution (300 mOsm, pH 7.4) was administered in 50-ml aliquots. Foals were carefully monitored for 4 days, then erythromycin base (25 mg/kg of body weight, PO, q 12 h) was given to foals of the treated group. After 4 days, foals were reanesthetized, and the same lung was relavaged. Cytologic examination ...
Study on the gastrointestinal parasite fauna of ponies in northern Germany.
Applied parasitology    December 1, 1996   Volume 37, Issue 4 239-244 
Cirak VY, Hermosilla C, Bauer C.Quantitative faecal and post-mortem examinations of 16 ponies, 1 to 2 1/2 years of age, originating from 3 farms in northern Germany were performed in February 1995 to determine the prevalence and intensity of gastrointestinal parasites in these animals. A total of 33 species of metazoan parasites was recovered: three tapeworm species (Anoplocephala perfoliata, A. magna, Paranoplocephala mamillana), Strongylus vulgaris, S. edentatus, small strongyles (including four Triodontophorus spp., Craterostomum acuticaudatum and 19 cyathostome species), Oxyuris equi, Parascaris equorum, Habronema majus ...
The effect of inflammation on the disposition of phenylbutazone in thoroughbred horses.
Journal of veterinary pharmacology and therapeutics    December 1, 1996   Volume 19, Issue 6 475-481 doi: 10.1111/j.1365-2885.1996.tb00085.x
Mills PC, Ng JC, Auer DE.The effect of inflammation on the disposition of phenylbutazone (PBZ) was investigated in Thoroughbred horses. An initial study (n = 5) in which PBZ (8.8 mg/kg) was injected intravenously twice, 5 weeks apart, suggested that the administration of PBZ would not affect the plasma kinetics of a subsequent dose. Two other groups of horses were given PBZ at either 8.8 mg/kg (n = 5) or 4.4 mg/kg (n = 4). Soft tissue inflammation was then induced by the injection of Freud's adjuvant and the administration of PBZ was repeated at a dose level equivalent to, but five weeks later than, the initial dose. ...
Equine synovial villi: distinctive structural organization of vasculature and novel nerve endings.
The Journal of veterinary medical science    December 1, 1996   Volume 58, Issue 12 1193-1204 doi: 10.1292/jvms.58.12_1193
Izumisawa Y, Yamaguchi M, Bertone AL, Tangkawattana P, Masty J, Yamashita K, Kotani T.The structural arrangement and cellular distribution of endothelial and lining cells of the synovial villi were studied in the equine palmar/plantar recess of the metacarpo- and metatarsophalangeal joints by light microscopy and electron microscopy. The extent and distribution of blood vessels varied with villous shape and length. The majority of vessels formed concentric circles in cross and longitudinal sections and probably are arranged in a convoluted, spiral or helical pattern. The villi do not contain smooth muscle cells or typical capillaries as observed in other organs. Under the elect...
Pharmacokinetics and pharmacodynamics of ketoprofen enantiomers in the horse.
Journal of veterinary pharmacology and therapeutics    December 1, 1996   Volume 19, Issue 6 466-474 doi: 10.1111/j.1365-2885.1996.tb00084.x
Landoni MF, Lees P.Pharmacokinetic and pharmacodynamic parameters were established for enantiomers of the non-steroidal anti-inflammatory drug (NSAID) ketoprofen (KTP), each administered separately at a dose level of 1.1 mg/kg to a group of six New Forest geldings, in a three-period cross-over study using a tissue cage model of inflammation. For both S(+)-and R(-)-KTP, penetration into tissue cage fluid (transudate) and inflamed tissue cage fluid (exudate) was rapid, and clearances from exudate and transudate were much slower than from plasma. AUC values were, therefore, higher for exudate and, to a lesser degre...
Alterations in equine guttural pouch morphology with head position: observations using a new technique for producing accurate casts.
The Anatomical record    December 1, 1996   Volume 246, Issue 4 579-584 doi: 10.1002/(SICI)1097-0185(199612)246:43.0.CO;2-L
Babptiste KE, Holladay SD, Freeman LE.An improved technique for preparing casts of the equine guttural pouch is described. The new technique is simplified and inexpensive in comparison to previous techniques and produces highly accurate anatomic models which are both durable and flexible. Methods: Silicone rubber bathtub caulk was injected into the guttural pouches of horse cadavers. The silicone was allowed to cure overnight, and casts were removed by dissection. The new technique was then used to evaluate changes in the shape of the guttural pouch with changes in head position. Results: With flexion of the atlanto-occipital join...
Measurement of extravascular lung water by the double indicator dilution method using heat and sodium in horses under general anesthesia.
The Journal of veterinary medical science    December 1, 1996   Volume 58, Issue 12 1205-1209 doi: 10.1292/jvms.58.12_1205
Ito S, Ishimaru M, Hobo S, Fujinaga T.Rapid infusion is believed to be harmful to the lung, however, the pathological status of pulmonary edema resulting from excessive fluid therapy in horses has not been clarified because the quantitative diagnosis of pulmonary edema is impossible. To evaluate the precision of the double indicator dilution method using heat and sodium in horses, which allows the quantitative diagnosis of pulmonary edema, we compared extravascular lung water volume measured using a lung water computer based on the theory of the double indicator dilution method with that determined by the direct method. The value ...
Effect of bethanechol or erythromycin on gastric emptying in horses.
American journal of veterinary research    December 1, 1996   Volume 57, Issue 12 1771-1775 
Ringger NC, Lester GD, Neuwirth L, Merritt AM, Vetro T, Harrison J.To investigate the prokinetic effect of bethanechol and erythromycin in the upper gastrointestinal tract of healthy horses by measuring the gastric emptying (GE) rate of a radioactive meal. Methods: 4 healthy adult horses. Methods: After food was withheld for 12 hours, horses were given 370 MBq of 99mTc-labeled sulfur colloid incorporated into egg albumen and 37 MBq of 111In-labeled diethyltriaminepentaacetic acid in 120 ml of water via nasogastric intubation. Intravenously administered treatments were 0.9% NaCl solution, erythromycin (0.1 or 1.0 mg/kg of body weight), or bethanechol (0.25 mg/...
Collagen fiber organization is related to mechanical properties and remodeling in equine bone. A comparison of two methods.
Journal of biomechanics    December 1, 1996   Volume 29, Issue 12 1515-1521 
Martin RB, Lau ST, Mathews PV, Gibson VA, Stover SM.We studied birefringence as an indicator of collagen fiber orientation in the diaphysis of the equine third metacarpal bone. We had previously shown that tissue from the lateral cortex of this bone is stronger monotonically, but less fatigue resistant, than tissue from the medial and dorsal regions. To learn whether collagen fiber orientation might play a role in this regional specialization, we tested three hypotheses using the same specimens: (1) collagen fiber orientation is regionally dependent; (2) remodeling changes collagen fiber orientation; (3) longitudinal collagen fibers correlate p...
The equine periodic paralysis Na+ channel mutation alters molecular transitions between the open and inactivated states.
The Journal of physiology    December 1, 1996   Volume 497 ( Pt 2), Issue Pt 2 349-364 doi: 10.1113/jphysiol.1996.sp021773
Hanna WJ, Tsushima RG, Sah R, McCutcheon LJ, Marban E, Backx PH.1. The Na+ channel mutation associated with equine hyperkalaemic periodic paralysis (HPP) affects a highly conserved phenylalanine residue in an unexplored region of the alpha-subunit. This mutation was introduced into the rat skeletal muscle Na+ channel gene at the corresponding location (i.e. F1412L) for functional expression and characterization in Xenopus oocytes. 2. In comparison with wild-type (WT) channels, equine HPP channels showed clear evidence for disruption of inactivation: increased time-to-peak current, slowed rates of whole-cell current decay, significant increases in sustained...
Distribution of penicillins into subcutaneous tissue chambers in ponies.
Journal of veterinary pharmacology and therapeutics    December 1, 1996   Volume 19, Issue 6 439-444 doi: 10.1111/j.1365-2885.1996.tb00080.x
Ensink JM, Klein WR, Barneveld A, Vulto AG, Van Miert AS, Tukker JJ.The distribution of penicillins into a tissue chamber implanted subcutaneously in ponies was studied. Ampicillin sodium (equivalent to 15 mg/kg ampicillin) was administered intravenously. Pivampicillin, a prodrug of ampicillin, was administered by nasogastric tube to fed ponies at a dose of 19.9 mg/kg (equivalent to 15 mg/kg ampicillin). Procaine penicillin G was administered intramuscularly at a dose of 12 mg/kg (equivalent to 12000 IU/kg). Six ponies were used for each medication. Antibiotic concentrations in plasma and tissue chamber fluid (TCF) were measured for 24 h after administration. ...
A 19 kDa protein secreted by the endometrium of the mare is a novel member of the lipocalin family.
The Biochemical journal    November 15, 1996   Volume 320 ( Pt 1), Issue Pt 1 137-143 doi: 10.1042/bj3200137
Crossett B, Allen WR, Stewart F.Large quantities of an unusual 19 kDa protein (p19) are secreted into the lumen of the uterus of the mare (Equus caballus) during the oestrous cycle and early pregnancy. p19 associates strongly with the acellular capsule that surrounds the young horse conceptus and is believed to be important in maintaining pregnancy. Here we report the complete cDNA sequence encoding p19, its expression patterns in horse tissues and a Southern blot analysis of the gene in horse DNA. The predicted amino acid sequence of the p19 cDNA demonstrated a signal peptide of 18 residues and a mature protein of 162 resid...
The role of ADP in endotoxin-induced equine platelet activation.
European journal of pharmacology    November 14, 1996   Volume 315, Issue 2 203-212 doi: 10.1016/s0014-2999(96)00637-1
Jarvis GE, Evans RJ, Heath MF.We have shown previously that endotoxin induces platelet aggregation in equine heparinised whole blood in a platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) dependent manner. ADP is an agonist of platelets and is present in platelet dense granules with ATP in high concentrations. An investigation was carried out to establish whether endotoxin-induced platelet activation was associated with release of platelet ATP and ADP. ADP-scavenging enzyme systems significantly inhibited endotoxin-induced aggregation. Plasma levels of adenine nucleotides were measured using ...
Use of diagnostic ultrasonography in horses with signs of acute abdominal pain.
Journal of the American Veterinary Medical Association    November 11, 1996   Volume 209, Issue 9 1597-1601 
Klohnen A, Vachon AM, Fischer AT.To evaluate the use of abdominal ultrasonography as a diagnostic tool in horses with signs of colic. Methods: Prospective study. Methods: 226 horses with signs of acute abdominal pain were compared to 20 clinical normal horses. Methods: The following were performed in horses with signs of colic: physical examination, CBC, abdominal fluid analysis, placement of a nasogastric tube to obtain gastric reflux, abdominal palpation per rectum, and ultrasonography of the abdomen. Results of ultrasonography were compared with the surgical, necropsy, or medical findings. Results: Ultrasonography of horse...
Variation of an epitope of keratan sulphate and total glycosaminoglycans in normal equine joints.
Equine veterinary journal    November 1, 1996   Volume 28, Issue 6 490-493 doi: 10.1111/j.2042-3306.1996.tb01622.x
Fuller CJ, Barr AR, Dieppe PA, Sharif M.An epitope of keratan sulphate (KS) and total glycosaminoglycans (GAG) were measured in synovial fluid samples from joints of 53 horses immediately following humane destruction. Internal examination of the joints post mortem ensured that there was no gross evidence of osteoarthritis or other joint disease. Joints sampled were distal interphalangeal (DIP), proximal interphalangeal (PIP), metacarpophalangeal (MCP), metatarsophalangeal (MTP), tarsometatarsal (TMT), tarsocrural (TC), femoropatellar (FP) and antebrachiocarpal (ABC) joints. The age of each horse was assessed by examination of the te...
Lead toxicosis in 2 horses: similarity to equine degenerative lower motor neuron disease.
Journal of veterinary internal medicine    November 1, 1996   Volume 10, Issue 6 420-423 doi: 10.1111/j.1939-1676.1996.tb02091.x
Sojka JE, Hope W, Pearson D.No abstract available
Natural superinfection of Parascaris equorum in a stall-confined orphan horse foal.
Veterinary parasitology    November 1, 1996   Volume 66, Issue 1-2 119-123 doi: 10.1016/s0304-4017(96)00996-x
Lyons ET, Swerczek TW, Tolliver SC, Drudge JH.At 4 days of age, an orphan horse foal born on a pasture was placed in a parasite-free stall. It was euthanized at 205 days of age and examination revealed numerous. Parascaris equorum specimens, ranging from 3.0 mm to adult size, indicating reinfections in the stall over an extended period of time. Initial infection of the foal was from stages on pasture. The foal had been observed repeatedly eating its own feces and the question is posed as to whether "recycling' of P. equorum eggs several times by coprophagy allowed time for embryonation and reinfection to occur.
Effect of omeprazole on sodium and potassium output in pentagastrin-stimulated equine gastric contents.
American journal of veterinary research    November 1, 1996   Volume 57, Issue 11 1640-1644 
Merritt AM, Burrow JA, Horbal MJ, Madison JB, Tran T.To better characterize the source of the large nonparietal secretory response to pentagastrin (PG) expressed in gastric contents of cannulated horses. Methods: Adult cross-bred horses: 4 geldings and 1 mare. Methods: Horses were prepared by surgical insertion of a silastic gastric cannula from which gastric contents after feed was withheld could be continuously collected by gravity drainage. During experiments, the horses were lightly restrained in stocks, the gastric cannula was opened, and a catheter was inserted into a jugular vein. Over the next 5 hours, gastric contents were collected in ...
Use of peanut agglutinin to assess the acrosomal status and the zona pellucida-induced acrosome reaction in stallion spermatozoa.
Journal of andrology    November 1, 1996   Volume 17, Issue 6 674-682 
Cheng FP, Fazeli A, Voorhout WF, Marks A, Bevers MM, Colenbrander B.Peanut agglutinin (PNA) was used to assess the sperm acrosomal status and the acrosome reaction during gamete interaction in the equine species. PNA exclusively binds to the outer acrosomal membrane of stallion spermatozoa, as was established by transmission electron microscopy. Fluorescein isothiocyanate-PNA (FITC-PNA) labeling was used to monitor sperm acrosomal changes during a prolonged incubation period of 24 hours and during a 2-hours incubation in the presence of 5 microM calcium ionophore A23187. In addition, after a 4-hours preincubation in SP-TALP medium, sperm samples were incubated...
Analysis of the equine tumor suppressor gene p53 in the normal horse and in eight cutaneous squamous cell carcinomas.
Cancer letters    October 1, 1996   Volume 107, Issue 1 125-130 doi: 10.1016/0304-3835(96)04359-5
Pazzi KA, Kraegel SA, Griffey SM, Theon AP, Madewell BR.Wild type equine p53 was amplified between exons 2 and 9 by the polymerase chain reaction using primers designed from conserved regions in other species. An 828 base pair region, corresponding to codons 25-313 of human p53, was sequenced in both directions. Human and equine amino acid sequences were 87% homologous in this region and 96% homologous in conserved domains II-V. Of eight equine cutaneous or mucocutaneous squamous cell carcinomas directly sequenced from exons 5-8, two had p53 point mutations resulting in single amino acid substitutions.
Mechanism of exercise-induced augmentation of lymphokine activated killer (LAK) cell activity in the horse.
Veterinary immunology and immunopathology    October 1, 1996   Volume 53, Issue 3-4 221-233 doi: 10.1016/S0165-2427(96)05610-3
Horohov DW, Keadle TL, Pourciau SS, Littlefield-Chabaud MA, Kamerling SG, Keowen ML, French DD, Melrose PA.Intense exercise affects various parameters of the immune system. The overall effect of exercise on immune function is dependent upon the physical condition of the subject, the intensity and duration of the exercise period, and the immune parameter assessed. Unconditioned horses subjected to a single bout of intensive exercise exhibit multiple alterations in immune function, including an augmentation of lymphokine activated killer (LAK) cell function. This increase in LAK cell activity is not due to an increase in circulating LAK precursors. While peripheral blood mononuclear cells from exerci...
Eosinophilic keratoconjunctivitis in seven horses.
Journal of the American Veterinary Medical Association    October 1, 1996   Volume 209, Issue 7 1283-1286 
Yamagata M, Wilkie DA, Gilger BC.Eosinophilic keratoconjunctivitis was diagnosed in 7 horses at The Ohio State University between 1976 and 1994. All horses had moderate-to-severe blepharospasm, chemosis, and conjunctival hyperemia; epiphora; and extensive yellow-to-white caseous mucoid discharge. Corneal ulcers associated with this disease were perilimbal and extended centrally. All ulcers were covered with a white necrotic plaque firmly attached to the underlying cornea. Other ophthalmic abnormalities were not detected. Corneal scrapings examined cytologically contained numerous eosinophils interspersed between epithelial ce...
Experimental inoculation of foals and pigs with an enterotoxigenic E. coli isolated from a foal.
Veterinary microbiology    October 1, 1996   Volume 52, Issue 3-4 249-257 doi: 10.1016/s0378-1135(96)80744-9
Holland RE, Grimes SD, Walker RD, Wilson RA.Hemolytic E. coli strain 807-13, O149:NM:K88(STb+, LT+), was isolated from the feces of a neonatal diarrheic foal. E. coli 807-13 was examined for adhesion to brush border membranes (BBM) from foals, adult horses and pigs, and its pathogenicity was assessed in neonatal foals and pigs. E. coli 807-13 did not adhere to equine BBM but adhered to pig BBM. It did not cause diarrhea nor did it colonize the intestinal epithelium of 3 colostrum-deprived and 3 suckled foals challenged at 24 h of age. Acute ulcerative gastritis and acute suppurative gastritis were observed in 2 colostrum-deprived challe...
Bioavailability of ketoprofen in horses after rectal administration.
Journal of veterinary pharmacology and therapeutics    October 1, 1996   Volume 19, Issue 5 359-363 doi: 10.1111/j.1365-2885.1996.tb00064.x
Corveleyn S, Deprez P, Van der Weken G, Baeyens W, Remon JP.Six healthy mares ranging in age from 6 to 12 years and weighing from 415 to 540 kg were used to determine the rectal bioavailability of ketoprofen. For the rectal administration, three different formulations, each containing 1 g of ketoprofen, were administered in a fatty and a hydrophilic suppository base and as a liquid suspension. An average elimination half-life of 1.3 h (+/-1.2) was found. The average value for the total plasma clearance (ClT) was 131.9mL/ min.kg (range 95-183.5). The volume of distribution Vd(area) was 255 mL/kg and the mean residence time (MRT) value was 0.47 h. After ...
Phorbol ester stimulation of equine macrophage cultures alters expression of equine infectious anemia virus.
Veterinary microbiology    October 1, 1996   Volume 52, Issue 3-4 209-221 doi: 10.1016/s0378-1135(96)00071-5
Sellon DC, Walker KM, Russell KE, Perry ST, Fuller FJ.Equine infectious anemia virus (EIAV) is a lentivirus that replicates predominantly in mature tissue macrophages. Viral expression is strongly influenced by the state of differentiation of the host cell. While blood monocytes can be infected, viral transcription is limited until the cell differentiates into a mature macrophage. Activation of mature macrophages infected with EIAV might also alter viral expression, presumably through binding of cellular transcription factors to viral nucleic acid sequences within the long terminal repeat (LTR). Using DNA amplification techniques, we compared LTR...
Experimental contact of bighorn sheep (Ovis canadensis) with horses and cattle, and comparison of neutrophil sensitivity to Pasteurella haemolytica cytotoxins.
Journal of wildlife diseases    October 1, 1996   Volume 32, Issue 4 594-602 doi: 10.7589/0090-3558-32.4.594
Foreyt WJ, Lagerquist JE.Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin-dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils. All isolates of P. haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils. Thirteen of 18 R haemolytica biotype A isolates were cytotoxic (> 50% neutrophi...
Excitatory prejunctional beta 2-adrenoceptor distribution within equine airway cholinergic nerves.
Respiration physiology    October 1, 1996   Volume 106, Issue 1 81-90 doi: 10.1016/0034-5687(96)00062-x
Zhang XY, Zhu FX, Robinson NE.We examined the effect of activation of beta 2-adrenoceptor (AR) by isoproterenol (ISO) on acetylcholine (ACh) release evoked by electrical field stimulation (EFS: 20 V, 0.5 Hz, 0.5 msec) from cholinergic nerves in five regions of equine airways. We also tested if the effect of ISO was dependent on epithelium or prostanoids by examining the effect of ISO on ACh release in the presence and absence of epithelium or cyclooxygenase inhibition. Trachealis strips or bronchial rings were preincubated for 60 min with 10(-7) M atropine, 10(-6) M neostigmine, and 10(-5) M guanethidine. The ACh amount wa...
Nucleologenesis and ribonucleic acid synthesis in preimplantation equine embryos.
Biology of reproduction    October 1, 1996   Volume 55, Issue 4 769-774 doi: 10.1095/biolreprod55.4.769
Grøndahl C, Hyttel P.The nucleolus is believed to be the active site of rRNA synthesis in all eukaryotic cells. In preimplantation embryos, the embryonic genome is apparently more or less silent up to a species-specific developmental stage at which a major burst of transcription occurs. Here we report on nucleologenesis and some ultrastructural aspects of the onset of RNA synthesis in equine embryos during in vivo development. The zygotes and embryos up to blastocyst stages were surgically recovered from normally cycling mares. Mares were induced to ovulate by treatment with 3000 IU hCG and inseminated 20 and 34 h...
Nitric oxide and exercise in the horse.
The Journal of physiology    September 15, 1996   Volume 495 ( Pt 3), Issue Pt 3 863-874 doi: 10.1113/jphysiol.1996.sp021638
Mills PC, Marlin DJ, Demoncheaux E, Scott C, Casas I, Smith NC, Higenbottam T.1. The effects of exercise on the production rate of nitric oxide (NO) in exhaled air (VNO) and the effects of inhaled NO (80 p.p.m.) on cardiovascular and respiratory parameters were investigated in five Throughbred horses. 2. The concentration of NO ([NO]) in exhaled air collected from within the nasal opening was lower when collected at a high flow rate of 80 l min-1 than at a low flow rate of 20 l min-1: when trotting at 3.7 m s-1 the values were 0.78 +/- 0.15 and 1.23 +/- 9.14 p.p.b., respectively, and when cantering at 9 m s-1 the values were 1.69 +/- 0.31 and 2.25 +/- 0.32 p.p.b., respe...