Abstract:SUMMARY
Caecal fluid samples collected 8 and 24 hours after carbohydrate overload were quantitatively compared to control samples in terms of aerobic and anaerobic bacteria. Concomitant increases in lactic acid‐producing bacteria and decreases in Gram negative bacteria were substantiated during the onset of acute laminitis. Progressive decreases in caecal fluid pH were also quantitated. Although endotoxin assays of caecal fluid and blood were not done, the caecal flora changes suggest its presence during the o… Show more
“…and Lactobacillus spp. (15,16). These changes often lead to the development of acute laminitis some 36 to 48 h later (14).…”
Section: Discussionmentioning
confidence: 99%
“…Gram-positive bacteria have been implicated in the pathogenesis of laminitis in the carbohydrate overload model since they have been shown to overgrow in the cecum of horses given corn starch by stomach tube (16). This overgrowth, particularly of streptococci and lactobacilli, was associated with lactic acid accumulation and the death of gram-negative bacteria leading to endotoxin release, both of which were postulated to be the cause of this condition (15,31).…”
Acute laminitis has been associated with the overgrowth of gram-positive bacteria within the equine hindgut, causing the release of factor(s) leading to ischemia-reperfusion of the digits. The products of fermentation which trigger acute laminitis are, as yet, unknown; however, vasoactive amines are possible candidates. The objectives of this study were to use an in vitro model of carbohydrate overload to study the change in populations of cecal streptococci and lactobacilli and to establish whether certain species of these bacteria were capable of producing vasoactive amines from amino acids. Cecal contents from 10 horses were divided into aliquots and incubated anaerobically with either corn starch or inulin (fructan; both at 1 g/100 ml). Samples were taken at 6-h intervals over a 24-h period for enumeration of streptococci, lactobacilli, and gram-negative anaerobes by a dilution method onto standard selective growth media. The effects of the antibiotic virginiamycin (1 mg/100 ml) and calcium hydrogen phosphate (CaHPO 4 ; 0.3 g/100 ml) were also examined. Fermentation of excess carbohydrate was associated with increases in numbers of streptococci and lactobacilli (2-to 3.5-log unit increases; inhibited by virginiamycin) but numbers of gram-negative anaerobes were not significantly affected. A screening agar technique followed by 16S rRNA gene sequence analysis enabled the identification of 26 different bacterial strains capable of producing one or more vasoactive amines. These included members of the species Streptococcus bovis and five different Lactobacillus spp. These data suggest that certain bacteria, whose overgrowth is associated with carbohydrate fermentation, are capable of producing vasoactive amines which may play a role in the pathogenesis of acute laminitis.Fermentation of carbohydrate in the hindgut has been recognized as one of the primary events associated with acute laminitis in the horse (14). The ability of the equine small intestine to digest starch may be overwhelmed if starch is present in excess (Ͼ0.4% body weight [35]), leading to its metabolism by cecal and colonic bacteria and the production of lactic acid. Fructans, the oligosaccharides present in grass which act as the storage form of carbohydrate, do not appear to be digested by the small intestine (21) and so are also substrates for fermentative bacteria. The variation of fructan levels in grasses under different climatic conditions has been suggested to account for the seasonal incidence of acute laminitis (A. C. Longland, A. J. Cairns, and M. O. Humphreys, Proc. 16th Equine Nutr. Physiol. Soc., p. 258-259, 1999).Gram-positive bacteria have been implicated in the pathogenesis of laminitis in the carbohydrate overload model since they have been shown to overgrow in the cecum of horses given corn starch by stomach tube (16). This overgrowth, particularly of streptococci and lactobacilli, was associated with lactic acid accumulation and the death of gram-negative bacteria leading to endotoxin release, both of which were postulated t...
“…and Lactobacillus spp. (15,16). These changes often lead to the development of acute laminitis some 36 to 48 h later (14).…”
Section: Discussionmentioning
confidence: 99%
“…Gram-positive bacteria have been implicated in the pathogenesis of laminitis in the carbohydrate overload model since they have been shown to overgrow in the cecum of horses given corn starch by stomach tube (16). This overgrowth, particularly of streptococci and lactobacilli, was associated with lactic acid accumulation and the death of gram-negative bacteria leading to endotoxin release, both of which were postulated to be the cause of this condition (15,31).…”
Acute laminitis has been associated with the overgrowth of gram-positive bacteria within the equine hindgut, causing the release of factor(s) leading to ischemia-reperfusion of the digits. The products of fermentation which trigger acute laminitis are, as yet, unknown; however, vasoactive amines are possible candidates. The objectives of this study were to use an in vitro model of carbohydrate overload to study the change in populations of cecal streptococci and lactobacilli and to establish whether certain species of these bacteria were capable of producing vasoactive amines from amino acids. Cecal contents from 10 horses were divided into aliquots and incubated anaerobically with either corn starch or inulin (fructan; both at 1 g/100 ml). Samples were taken at 6-h intervals over a 24-h period for enumeration of streptococci, lactobacilli, and gram-negative anaerobes by a dilution method onto standard selective growth media. The effects of the antibiotic virginiamycin (1 mg/100 ml) and calcium hydrogen phosphate (CaHPO 4 ; 0.3 g/100 ml) were also examined. Fermentation of excess carbohydrate was associated with increases in numbers of streptococci and lactobacilli (2-to 3.5-log unit increases; inhibited by virginiamycin) but numbers of gram-negative anaerobes were not significantly affected. A screening agar technique followed by 16S rRNA gene sequence analysis enabled the identification of 26 different bacterial strains capable of producing one or more vasoactive amines. These included members of the species Streptococcus bovis and five different Lactobacillus spp. These data suggest that certain bacteria, whose overgrowth is associated with carbohydrate fermentation, are capable of producing vasoactive amines which may play a role in the pathogenesis of acute laminitis.Fermentation of carbohydrate in the hindgut has been recognized as one of the primary events associated with acute laminitis in the horse (14). The ability of the equine small intestine to digest starch may be overwhelmed if starch is present in excess (Ͼ0.4% body weight [35]), leading to its metabolism by cecal and colonic bacteria and the production of lactic acid. Fructans, the oligosaccharides present in grass which act as the storage form of carbohydrate, do not appear to be digested by the small intestine (21) and so are also substrates for fermentative bacteria. The variation of fructan levels in grasses under different climatic conditions has been suggested to account for the seasonal incidence of acute laminitis (A. C. Longland, A. J. Cairns, and M. O. Humphreys, Proc. 16th Equine Nutr. Physiol. Soc., p. 258-259, 1999).Gram-positive bacteria have been implicated in the pathogenesis of laminitis in the carbohydrate overload model since they have been shown to overgrow in the cecum of horses given corn starch by stomach tube (16). This overgrowth, particularly of streptococci and lactobacilli, was associated with lactic acid accumulation and the death of gram-negative bacteria leading to endotoxin release, both of which were postulated t...
“…A lowered gut pH environment favors the growth of species such as Streptococcus bovis and L. acidophilus. In the horse, low hindgut pH decreased numbers of cellulolytic and hemicellulolytic bacteria (Garner et al, 1978, Goodson et al, 1988, which significantly decreased forage utilization McLean et al, 2000;. A neutral fecal pH around 7.0 in the horse is known to support fiber digestion , however a cecal pH drop from 6.7 to 6.4 was due to altered fermentation patterns and microbial ecology in the hindgut of the horse fed high-starch concentrates .…”
This research evaluated the viability of lactic acid bacteria (LAB) intended for in vivo application as direct-fed microbial (DFM) supplements in two experiments during feed processing (Exp. 1) and storage (Exp. 2) and determined the efficacy of DFM on the digestibility and hindgut fermentation of horses during and after an abrupt increase in starch (Exp. 3). In Exp. 1, lactobacilli survived feed processing and a commercial enumeration method was validated. In Exp. 2, viable colony forming units of LAB were assessed and remained viable during 12 weeks of storage. Controls in both experiments had high levels of naturally-occurring bacteria present. In Exp. 3, a high-starch concentrate caused fecal pH to decrease, and fecal propionate and digestibility of many nutrients to increase. The DFM induced minimal improvements in digestibility or fermentation parameters and data provided no clear evidence to support the use of a multiple versus a single strain DFM preparation.
“…There is a resultant decrease in hindgut pH, which causes death of Gram negative bacteria and liberation of endotoxin (Garner et al 1977;Garner et al 1978). There is an increase in intestinal permeability and destruction of the caecal mucosal barrier (Krueger et al 1986;.…”
Section: Theories On Laminitis Pathogenesismentioning
confidence: 99%
“…In this model laminitis is induced by nasogastric administration of a bolus dose of corn starch and wood flour or rye flour. The excess starch rapidly ferments within the hindgut with a subsequent increase in Gram positive bacteria and lactic acidemia (Garner et al 1978). …”
Section: Theories On Laminitis Pathogenesismentioning
Laminitis, a serious and debilitating disease of the equine foot, is a significant cause of wastage in the equine industry. There is no scientifically validated treatment and laminitis prophylaxis is currently limited to digital cryotherapy. Several key steps in laminitis pathophysiology have been elucidated, resulting in identification of drugs that may prevent laminitis (anti-laminitis drugs (ALDs)). Many of these pharmaceuticals are not suitable for systemic delivery but may be amenable to either local or nanoparticle delivery mechanisms.The aim of this thesis was to evaluate local, systemic and nanoparticle delivery mechanisms, using the matrix metalloproteinase (MMP) inhibitor marimastat, to establish a method of drug delivery that yields sustained therapeutic lamellar concentrations for experimental and potential clinical use. In order to establish marimastat concentrations necessary for inhibition of lamellar MMPs, zymography was performed using lamellar homogenates incubated in increasing concentrations of marimastat. Based on this assay, target marimastat concentrations of 177 ng/mL were set (the concentration necessary to inhibit 90% of lamellar MMP-2 and MMP-9).
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