This study focuses on comprehensive characterization of the venom proteome of the beaked sea snak... more This study focuses on comprehensive characterization of the venom proteome of the beaked sea snake (Hydrophis schistosus) from Songkhla Lake, Thailand. H. schistosus can be considered as the deadliest sea snake commonly found in the Pacific and Indian oceans. Their envenomation causes muscular paralysis and rhabdomyolysis. To develop effective treatment for this snakebite, it is necessary to understand the detailed venom composition. In this study, multiple mass spectrometry-based approaches were employed. Bottom-up proteomics revealed that tryptic digestion in-solution provided a higher number of toxin proteins identified and a larger sequence coverage, compared to in-gel digestion. In addition, a venom gland transcriptome-derived database was constructed and used as a reference, which 43 known and novel toxin proteins were identified using this database and the UniProtKB. Three-finger toxin and phospholipase A2 were shown to be top two most abundant protein families. Minor compositions included other toxin families and a number of non-toxin proteins. Moreover, a hybrid de novo sequencing was performed to enhance identification of the small proteins/peptides. Using non-digested samples, there were 46 predicted toxin peptides. The finding from this study could lead to a better understanding in pathological effects of the snakebite and the future development of effective antivenoms. SIGNIFICANCE: This study provides a better understanding of the venom proteome composition of the beaked sea snake (H. schistosus) found in the Gulf of Thailand, using a combination of different sample preparation techniques, Serpentes protein database searching, transcriptome-derived protein database searching, and a hybrid de novo peptide sequencing strategy. It revealed 13 toxin protein families and novel proteins in the beaked sea snake venom including new species of phospholipase A2s (PLA2s) and three-finger toxins (3FTxs). It could serve as a basis for the development of snakebite treatments and for the discovery of novel pharmaceutical drugs from the toxin peptides.
วารสารเภสัชวิทยา (Thai Journal of Pharmacology), 1985
In this study, we assessed the effects of RVV on renal hemodynamics, urinary enzymes, plasma and ... more In this study, we assessed the effects of RVV on renal hemodynamics, urinary enzymes, plasma and urinary Tx in 7 dogs during and after a 40-min intrarenal arterial infusion of RVV (1.25 μg/kg/min). All parameters were determined before and at 20, 40 and 60 min after the beginning of RVV. The urinary enzymes measured included N-acetyl-s-D-glucosaminidase (NAG). γ -glutamyl, transpeptidase ( γ -GT), alanine-aminopeptidase(AAP) and s-galactosidase (GAL). Plasma and urinary Tx were measured by radioimmunoassay after extraction and purification steps. As compared with the control stage, significant change was observed for a decrease in the mean arterial pressure (MAP) during the first 20 min after RVV administration (128.4 ± 7.7 vs. 118 ± 10.2 mm Hg. mean±SE, p < 0 .025) as well as the fractional excretion of sodium (), potassium ( ), and chloride () (p <0.05). The heart rate (HR), renal blood flow (RBF) and glomerular tiltration rate (GFR) did·not change significantly for all period of observation. Urine flow (V) increased by approximately 15.4-65.4% in 5 dogs and decreased 6.6-32.9% in 2 dogs. increased significantly 40 min after the commencement of RVV infusion (727 ±301 VS. 1484 ± 386 units/min, p < 0.005). A 7-to 9-fold elevation of was detected for all periods of observation but was significantly increased during the first 20 min after RVV infusion This study showed : (1) The changes of RBF and GFR could not explain the rise of V, excretion of electrolytes and . They should rather be accounted for by direct tubular dysfunction (2) Plasma increased in parallel with suggesting an augmented synthesis of and/or disturbance of active reabsorption of by tubular cells.(3) The elevation of NAG indicated direct nephrotoxicity of RVV.
Background and Aim: African horse sickness (AHS) is a non-contagious, high mortality, and insect-... more Background and Aim: African horse sickness (AHS) is a non-contagious, high mortality, and insect-borne disease caused by a double-stranded RNA virus from the genus Orbivirus. The study aimed to develop inactivated vaccines serotype 1 inactivated AHS vaccine (IAV) and to compare the effect of IAV on antibody responses in young naïve horses and adult horses pre-immunized with live-attenuated AHS virus (AHSV) serotypes 1, 3, and 4 live-attenuated vaccine (LAV). Materials and Methods: A total of 27 horses were vaccinated in two trials. Twelve AHS naïve young horses and 15 adult horses were divided into three groups of 4 and 5 horses each, respectively. Horses in control Group 1 were treated with phosphate-buffered saline. Horses in Group 2 were subcutaneously vaccinated with 2 mL of formulated IAV with 10% Gel 01™ (Seppic, France) on day 0 and horses in Group 3 were subcutaneously vaccinated with 2 mL of IAV on day 0 and a booster on day 28. The IAV vaccine was prepared by isolating the...
Background and Aim: Activation of breathing, the hypothalamic-pituitary-adrenal (HPA) axis, and p... more Background and Aim: Activation of breathing, the hypothalamic-pituitary-adrenal (HPA) axis, and plasma antioxidant defense are adaptive mechanisms in lactating dairy goats fed during the summer season. However, an excess of these responses can interfere with the gas exchange. This study aimed to investigate the effect of natural high ambient temperature (HTa) on blood gas parameters and their relation to the HPA axis and antioxidant defense. Materials and Methods: Six mid-lactating goats were included in this study and were fed in individual pens for 2 weeks. The data on ambient conditions, physiological responses, and blood chemistry were measured for two sampling days (D7 and D14), 1 week apart during the late summer season. On this two-sampling day, the main physiological responses to HTa, including respiration rate (RR), rectal temperature (Tr), blood gas, and blood chemistry, were measured in the morning and afternoon. Results: Goats from both D7 and D14 increased RR and Tr sig...
Journal of Veterinary Pharmacology and Therapeutics, 1993
ABSTRACT Four dogs were given 5 mg/kg body weight enrofloxacin intravenously (i.v.) and orally (p... more ABSTRACT Four dogs were given 5 mg/kg body weight enrofloxacin intravenously (i.v.) and orally (p.o.) in a cross-over study. Plasma concentrations of the active ingredient enrofloxacin and its main metabolite ciprofloxacin were determined by a reversed phase liquid chromatographic method. Pharmacokinetic parameters of both substances were calculated by use of statistical moments and were compared to those of enrofloxacin described in the veterinary literature. Mean enrofloxacin t1/2 lambda z was 2.4 h, mean Cls was 27.1 ml/min.kg, and mean Vss was 7.0 l/kg. After i.v. and p.o. administration, concentrations of ciprofloxacin exceeding minimal inhibitory concentrations of several microorganisms were reached (Cmax = 0.2 microgram/ml, tmax = 2.2 h after intravenous administration; Cmax = 0.2 microgram/ml, tmax = 3.6 h after oral administration). A considerable part of the antimicrobial activity is due to ciprofloxacin, the main metabolite of enrofloxacin.
Envenoming by Russells viper causes a broad spectrum of renal impairment. Renal failure is an imp... more Envenoming by Russells viper causes a broad spectrum of renal impairment. Renal failure is an important complication in patients bitten by Russells viper. Experimental work in animals and in vitro has elucidated pathophysiological mechanisms that contribute to life threatening complications and have suggested possibilities for therapeutic intervention. The evidence in experimental animals regarding mechanisms of venom action in relation to changes in either extrarenal or intrarenal factors is presented. The cardiovascular system and renal hemodynamics are affected by venom. Reductions of renal function including renal hemodynamics are associated directly with changes in general circulation during envenomation. Possible endogenous mechanisms for releasing the hormone inducing renal vasoconstriction after envenomation are evident. Hormonal factor such as the catecholamine, prostaglandin and renin angiotensin systems induce these changes. Direct nephrotoxicity of venom action is studie...
Background and Aim: Milk synthesis by the mammary gland is negatively influenced in part by high ... more Background and Aim: Milk synthesis by the mammary gland is negatively influenced in part by high ambient temperature (AT). This study aimed to clarify the pathway of intramammary glucose utilization involved in mediating lactose synthesis during treatment with somatotropin under housing with misters and fans. Materials and Methods: A single subcutaneous injection of 500 mg of recombinant bovine somatotropin (rbST) was administered 3 times once every 14 days to 87.5% crossbred Holstein cattle in early-/mid-/late lactation, under housing in a normal shaded barn and in a shaded barn with a mist-fan cooling system. Results: The milk yields of the cooled cows tended to increase compared with those of uncooled cows and exhibited more potentiated effects in response to rbST treatment, coinciding with increases in mammary plasma flow and glucose uptake, but not in the mammary extraction of glucose. Treatment with rbST in the cooled cows resulted in a greater increase in the milk glucose con...
Background and Aim: Metabolism and environment are closely related. Under high ambient temperatur... more Background and Aim: Metabolism and environment are closely related. Under high ambient temperature (HTa), dairy cows may have different energy metabolism during summer and winter. The present study was carried out to investigate the effect of HTa on the milk yield and blood concentration of beta-hydroxybutyrate (BHBA) and glucose at the herd level. Materials and Methods: One large dairy farm in Thailand with more than 100 crossbred Holstein cows milked each month was selected. The first experiment was performed on non-lactating cows to determine the normal daily concentrations of blood BHBA and glucose. Under the HTa condition, there was no significant change in blood BHBA and glucose concentrations. The second experiment was performed using a prospective cohort clinical design to demonstrate the seasonal effect on milk yield and blood BHBA as an indication of energy metabolism at the herd level. Results: The temperature and humidity index for the winter (78.1±0.5) and summer (83.4±...
An experiment was conducted to examine the effect of moonlight on the time spent grazing and wall... more An experiment was conducted to examine the effect of moonlight on the time spent grazing and wallowing by buffalo heifers during the late winter, summer and rainy seasons (February to December) in Thailand. Eight swamp buffalo heifer s were stocked continuously on improved grass-dominant pasture. Grazing time was determined by visual observation at 10-minute intervals over ten 24-h periods during both the full and the new phase s of the moon. Pedometers fitted to the buffaloes were used to measure walking activity during the measurement periods. Parallel curve analysis showed that during nights with a full moon, eating time increased by 80 minutes longer (P<0.001) compared with when there was a new moon. However, the total grazing time during 24 h was not significantly affected by the phases of the moon, the mean time being 484 ±16.8 min d-1. During the first 5 months, the buffaloes walked significantly further during 24 hours, when there was a full moon compared with when there ...
Thermal environment is regarded as the prime factor governing the production because, both the fe... more Thermal environment is regarded as the prime factor governing the production because, both the feed consumption and heat production are related closely to the thermal environment. Therefore, heat stress is caused by a combination of environmental and animal factors that result in reduced performance as the animal attempts to cool itself. In this review, the physiological reactions of bodily functions during heat exposure in poultry are described. Thermal panting, the marked response of poultry to heat stress, is compensatory behavior to permit a high rate of evaporative cooling from the respiratory tract. Cardiorespiratory responses to thermal stress show different patterns among birds that are experienced or inexperienced with high ambient temperature. The effect of heat stress on changes of body fluids and renal function of poultry are reviewed. A number of methods to reduce heat stress on poultry production are presented along with various management considerations, including wat...
Acute kidney injury (AKI) following Eastern Russell’s viper (Daboia siamensis) envenoming is a si... more Acute kidney injury (AKI) following Eastern Russell’s viper (Daboia siamensis) envenoming is a significant symptom in systemically envenomed victims. A number of venom components have been identified as causing the nephrotoxicity which leads to AKI. However, the precise mechanism of nephrotoxicity caused by these toxins is still unclear. In the present study, we purified two proteins from D. siamensis venom, namely RvPLA2 and RvMP. Protein identification using LCMS/MS confirmed the identity of RvPLA2 to be snake venom phospholipase A2 (SVPLA2) from Thai D. siamensis venom, whereas RvMP exhibited the presence of a factor X activator with two subunits. In vitro and in vivo pharmacological studies demonstrated myotoxicity and histopathological changes of kidney, heart, and spleen. RvPLA2 (3–10 µg/mL) caused inhibition of direct twitches of the chick biventer cervicis muscle preparation. After administration of RvPLA2 or RvMP (300 µg/kg, i.p.) for 24 h, diffuse glomerular congestion and...
This study focuses on comprehensive characterization of the venom proteome of the beaked sea snak... more This study focuses on comprehensive characterization of the venom proteome of the beaked sea snake (Hydrophis schistosus) from Songkhla Lake, Thailand. H. schistosus can be considered as the deadliest sea snake commonly found in the Pacific and Indian oceans. Their envenomation causes muscular paralysis and rhabdomyolysis. To develop effective treatment for this snakebite, it is necessary to understand the detailed venom composition. In this study, multiple mass spectrometry-based approaches were employed. Bottom-up proteomics revealed that tryptic digestion in-solution provided a higher number of toxin proteins identified and a larger sequence coverage, compared to in-gel digestion. In addition, a venom gland transcriptome-derived database was constructed and used as a reference, which 43 known and novel toxin proteins were identified using this database and the UniProtKB. Three-finger toxin and phospholipase A2 were shown to be top two most abundant protein families. Minor compositions included other toxin families and a number of non-toxin proteins. Moreover, a hybrid de novo sequencing was performed to enhance identification of the small proteins/peptides. Using non-digested samples, there were 46 predicted toxin peptides. The finding from this study could lead to a better understanding in pathological effects of the snakebite and the future development of effective antivenoms. SIGNIFICANCE: This study provides a better understanding of the venom proteome composition of the beaked sea snake (H. schistosus) found in the Gulf of Thailand, using a combination of different sample preparation techniques, Serpentes protein database searching, transcriptome-derived protein database searching, and a hybrid de novo peptide sequencing strategy. It revealed 13 toxin protein families and novel proteins in the beaked sea snake venom including new species of phospholipase A2s (PLA2s) and three-finger toxins (3FTxs). It could serve as a basis for the development of snakebite treatments and for the discovery of novel pharmaceutical drugs from the toxin peptides.
วารสารเภสัชวิทยา (Thai Journal of Pharmacology), 1985
In this study, we assessed the effects of RVV on renal hemodynamics, urinary enzymes, plasma and ... more In this study, we assessed the effects of RVV on renal hemodynamics, urinary enzymes, plasma and urinary Tx in 7 dogs during and after a 40-min intrarenal arterial infusion of RVV (1.25 μg/kg/min). All parameters were determined before and at 20, 40 and 60 min after the beginning of RVV. The urinary enzymes measured included N-acetyl-s-D-glucosaminidase (NAG). γ -glutamyl, transpeptidase ( γ -GT), alanine-aminopeptidase(AAP) and s-galactosidase (GAL). Plasma and urinary Tx were measured by radioimmunoassay after extraction and purification steps. As compared with the control stage, significant change was observed for a decrease in the mean arterial pressure (MAP) during the first 20 min after RVV administration (128.4 ± 7.7 vs. 118 ± 10.2 mm Hg. mean±SE, p < 0 .025) as well as the fractional excretion of sodium (), potassium ( ), and chloride () (p <0.05). The heart rate (HR), renal blood flow (RBF) and glomerular tiltration rate (GFR) did·not change significantly for all period of observation. Urine flow (V) increased by approximately 15.4-65.4% in 5 dogs and decreased 6.6-32.9% in 2 dogs. increased significantly 40 min after the commencement of RVV infusion (727 ±301 VS. 1484 ± 386 units/min, p < 0.005). A 7-to 9-fold elevation of was detected for all periods of observation but was significantly increased during the first 20 min after RVV infusion This study showed : (1) The changes of RBF and GFR could not explain the rise of V, excretion of electrolytes and . They should rather be accounted for by direct tubular dysfunction (2) Plasma increased in parallel with suggesting an augmented synthesis of and/or disturbance of active reabsorption of by tubular cells.(3) The elevation of NAG indicated direct nephrotoxicity of RVV.
Background and Aim: African horse sickness (AHS) is a non-contagious, high mortality, and insect-... more Background and Aim: African horse sickness (AHS) is a non-contagious, high mortality, and insect-borne disease caused by a double-stranded RNA virus from the genus Orbivirus. The study aimed to develop inactivated vaccines serotype 1 inactivated AHS vaccine (IAV) and to compare the effect of IAV on antibody responses in young naïve horses and adult horses pre-immunized with live-attenuated AHS virus (AHSV) serotypes 1, 3, and 4 live-attenuated vaccine (LAV). Materials and Methods: A total of 27 horses were vaccinated in two trials. Twelve AHS naïve young horses and 15 adult horses were divided into three groups of 4 and 5 horses each, respectively. Horses in control Group 1 were treated with phosphate-buffered saline. Horses in Group 2 were subcutaneously vaccinated with 2 mL of formulated IAV with 10% Gel 01™ (Seppic, France) on day 0 and horses in Group 3 were subcutaneously vaccinated with 2 mL of IAV on day 0 and a booster on day 28. The IAV vaccine was prepared by isolating the...
Background and Aim: Activation of breathing, the hypothalamic-pituitary-adrenal (HPA) axis, and p... more Background and Aim: Activation of breathing, the hypothalamic-pituitary-adrenal (HPA) axis, and plasma antioxidant defense are adaptive mechanisms in lactating dairy goats fed during the summer season. However, an excess of these responses can interfere with the gas exchange. This study aimed to investigate the effect of natural high ambient temperature (HTa) on blood gas parameters and their relation to the HPA axis and antioxidant defense. Materials and Methods: Six mid-lactating goats were included in this study and were fed in individual pens for 2 weeks. The data on ambient conditions, physiological responses, and blood chemistry were measured for two sampling days (D7 and D14), 1 week apart during the late summer season. On this two-sampling day, the main physiological responses to HTa, including respiration rate (RR), rectal temperature (Tr), blood gas, and blood chemistry, were measured in the morning and afternoon. Results: Goats from both D7 and D14 increased RR and Tr sig...
Journal of Veterinary Pharmacology and Therapeutics, 1993
ABSTRACT Four dogs were given 5 mg/kg body weight enrofloxacin intravenously (i.v.) and orally (p... more ABSTRACT Four dogs were given 5 mg/kg body weight enrofloxacin intravenously (i.v.) and orally (p.o.) in a cross-over study. Plasma concentrations of the active ingredient enrofloxacin and its main metabolite ciprofloxacin were determined by a reversed phase liquid chromatographic method. Pharmacokinetic parameters of both substances were calculated by use of statistical moments and were compared to those of enrofloxacin described in the veterinary literature. Mean enrofloxacin t1/2 lambda z was 2.4 h, mean Cls was 27.1 ml/min.kg, and mean Vss was 7.0 l/kg. After i.v. and p.o. administration, concentrations of ciprofloxacin exceeding minimal inhibitory concentrations of several microorganisms were reached (Cmax = 0.2 microgram/ml, tmax = 2.2 h after intravenous administration; Cmax = 0.2 microgram/ml, tmax = 3.6 h after oral administration). A considerable part of the antimicrobial activity is due to ciprofloxacin, the main metabolite of enrofloxacin.
Envenoming by Russells viper causes a broad spectrum of renal impairment. Renal failure is an imp... more Envenoming by Russells viper causes a broad spectrum of renal impairment. Renal failure is an important complication in patients bitten by Russells viper. Experimental work in animals and in vitro has elucidated pathophysiological mechanisms that contribute to life threatening complications and have suggested possibilities for therapeutic intervention. The evidence in experimental animals regarding mechanisms of venom action in relation to changes in either extrarenal or intrarenal factors is presented. The cardiovascular system and renal hemodynamics are affected by venom. Reductions of renal function including renal hemodynamics are associated directly with changes in general circulation during envenomation. Possible endogenous mechanisms for releasing the hormone inducing renal vasoconstriction after envenomation are evident. Hormonal factor such as the catecholamine, prostaglandin and renin angiotensin systems induce these changes. Direct nephrotoxicity of venom action is studie...
Background and Aim: Milk synthesis by the mammary gland is negatively influenced in part by high ... more Background and Aim: Milk synthesis by the mammary gland is negatively influenced in part by high ambient temperature (AT). This study aimed to clarify the pathway of intramammary glucose utilization involved in mediating lactose synthesis during treatment with somatotropin under housing with misters and fans. Materials and Methods: A single subcutaneous injection of 500 mg of recombinant bovine somatotropin (rbST) was administered 3 times once every 14 days to 87.5% crossbred Holstein cattle in early-/mid-/late lactation, under housing in a normal shaded barn and in a shaded barn with a mist-fan cooling system. Results: The milk yields of the cooled cows tended to increase compared with those of uncooled cows and exhibited more potentiated effects in response to rbST treatment, coinciding with increases in mammary plasma flow and glucose uptake, but not in the mammary extraction of glucose. Treatment with rbST in the cooled cows resulted in a greater increase in the milk glucose con...
Background and Aim: Metabolism and environment are closely related. Under high ambient temperatur... more Background and Aim: Metabolism and environment are closely related. Under high ambient temperature (HTa), dairy cows may have different energy metabolism during summer and winter. The present study was carried out to investigate the effect of HTa on the milk yield and blood concentration of beta-hydroxybutyrate (BHBA) and glucose at the herd level. Materials and Methods: One large dairy farm in Thailand with more than 100 crossbred Holstein cows milked each month was selected. The first experiment was performed on non-lactating cows to determine the normal daily concentrations of blood BHBA and glucose. Under the HTa condition, there was no significant change in blood BHBA and glucose concentrations. The second experiment was performed using a prospective cohort clinical design to demonstrate the seasonal effect on milk yield and blood BHBA as an indication of energy metabolism at the herd level. Results: The temperature and humidity index for the winter (78.1±0.5) and summer (83.4±...
An experiment was conducted to examine the effect of moonlight on the time spent grazing and wall... more An experiment was conducted to examine the effect of moonlight on the time spent grazing and wallowing by buffalo heifers during the late winter, summer and rainy seasons (February to December) in Thailand. Eight swamp buffalo heifer s were stocked continuously on improved grass-dominant pasture. Grazing time was determined by visual observation at 10-minute intervals over ten 24-h periods during both the full and the new phase s of the moon. Pedometers fitted to the buffaloes were used to measure walking activity during the measurement periods. Parallel curve analysis showed that during nights with a full moon, eating time increased by 80 minutes longer (P<0.001) compared with when there was a new moon. However, the total grazing time during 24 h was not significantly affected by the phases of the moon, the mean time being 484 ±16.8 min d-1. During the first 5 months, the buffaloes walked significantly further during 24 hours, when there was a full moon compared with when there ...
Thermal environment is regarded as the prime factor governing the production because, both the fe... more Thermal environment is regarded as the prime factor governing the production because, both the feed consumption and heat production are related closely to the thermal environment. Therefore, heat stress is caused by a combination of environmental and animal factors that result in reduced performance as the animal attempts to cool itself. In this review, the physiological reactions of bodily functions during heat exposure in poultry are described. Thermal panting, the marked response of poultry to heat stress, is compensatory behavior to permit a high rate of evaporative cooling from the respiratory tract. Cardiorespiratory responses to thermal stress show different patterns among birds that are experienced or inexperienced with high ambient temperature. The effect of heat stress on changes of body fluids and renal function of poultry are reviewed. A number of methods to reduce heat stress on poultry production are presented along with various management considerations, including wat...
Acute kidney injury (AKI) following Eastern Russell’s viper (Daboia siamensis) envenoming is a si... more Acute kidney injury (AKI) following Eastern Russell’s viper (Daboia siamensis) envenoming is a significant symptom in systemically envenomed victims. A number of venom components have been identified as causing the nephrotoxicity which leads to AKI. However, the precise mechanism of nephrotoxicity caused by these toxins is still unclear. In the present study, we purified two proteins from D. siamensis venom, namely RvPLA2 and RvMP. Protein identification using LCMS/MS confirmed the identity of RvPLA2 to be snake venom phospholipase A2 (SVPLA2) from Thai D. siamensis venom, whereas RvMP exhibited the presence of a factor X activator with two subunits. In vitro and in vivo pharmacological studies demonstrated myotoxicity and histopathological changes of kidney, heart, and spleen. RvPLA2 (3–10 µg/mL) caused inhibition of direct twitches of the chick biventer cervicis muscle preparation. After administration of RvPLA2 or RvMP (300 µg/kg, i.p.) for 24 h, diffuse glomerular congestion and...
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