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Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12) Полный текст
2017
Botha, Christo J.(University of Pretoria Department of Paraclinical Sciences) | Clift, Sarah J.(University of Pretoria Department of Paraclinical Sciences) | Ferreira, Gezina C.H.(University of Pretoria Department of Paraclinical Sciences) | Masango, Mxolisi G.(Agricultural Research Council-Onderstepoort Veterinary Institute Food, Feed and Veterinary Public Health)
Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12) Полный текст
2017
Botha, Christo J.(University of Pretoria Department of Paraclinical Sciences) | Clift, Sarah J.(University of Pretoria Department of Paraclinical Sciences) | Ferreira, Gezina C.H.(University of Pretoria Department of Paraclinical Sciences) | Masango, Mxolisi G.(Agricultural Research Council-Onderstepoort Veterinary Institute Food, Feed and Veterinary Public Health)
Geigeria poisoning in sheep, locally known as 'vermeersiekte', is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in 'vermeersiekte' and to elucidate the subcellular effects of these myotoxins on cultured myoblasts.
Показать больше [+] Меньше [-]Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12) Полный текст
2017
Christo J. Botha | Sarah J. Clift | Gezina C.H. Ferreira | Mxolisi G. Masango
Geigeria poisoning in sheep, locally known as ‘vermeersiekte’, is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in ‘vermeersiekte’ and to elucidate the subcellular effects of these myotoxins on cultured myoblasts.
Показать больше [+] Меньше [-]Assessment of a portable lactate meter for field use in the white rhinoceros (Ceratotherium simum) Полный текст
2017
Cole, Georgina C.(University of Pretoria Department of Companion Animal Clinical Studies) | Tordiffe, Adrian S.W.(University of Pretoria Department of Paraclinical Sciences) | Steenkamp, Gerhard(University of Pretoria Department of Companion Animal Clinical Studies)
Assessment of a portable lactate meter for field use in the white rhinoceros (Ceratotherium simum) Полный текст
2017
Cole, Georgina C.(University of Pretoria Department of Companion Animal Clinical Studies) | Tordiffe, Adrian S.W.(University of Pretoria Department of Paraclinical Sciences) | Steenkamp, Gerhard(University of Pretoria Department of Companion Animal Clinical Studies)
Blood lactate is a predictor of mortality in critically ill humans and animals. Handheld lactate meters have the potential to be used in the field to evaluate the condition of severely injured rhinoceroses but have not been compared with laboratory-based methods. Agreement between a handheld lactate meter and a laboratory method was assessed, as was the stability of rhino blood lactate in the anticoagulant sodium fluoride/potassium oxalate (fluoride/oxalate). Blood samples were obtained from 53 white rhinos that had been immobilised for management reasons. Lactate was measured by means of a handheld meter using whole blood in heparin (WBHEP), whole blood in fluoride/oxalate (WBFO) and fluoride/oxalate plasma (PFO). Results were recorded in both blood (BL) and plasma (PL) modes and compared to an established laboratory method for measuring plasma lactate. To assess the stability of lactate over time, blood lactate in fluoride/oxalate was measured on the handheld meter at intervals for up to 91 h. Agreement was best using WBFO in PL mode, with small bias (−0.16), tight 95% limits of agreement (LOA) (−1.46, 1.14) and a Pc (95% CI) of 0.97 (0.92, 0.99). The agreement was improved for all sample types when using the PL mode compared to the blood lactate (BL) mode. Blood lactate was stable in fluoride/oxalate for 91 h, with a mean change from baseline of 0.15 (−0.178, 0.478) mmol/L (mean, 95% CI). The handheld meter was found to be suitable for field use in white rhinos but provided more reliable results with the device in PL mode. Furthermore, rhino blood lactate was found to be stable in fluoride/oxalate for as long as 3 days.
Показать больше [+] Меньше [-]Alterations in conjunctival bacteria and antimicrobial susceptibility during topical administration of ofloxacin after cataract surgery in dogs Полный текст
2017
OBJECTIVE To evaluate changes in conjunctival bacteria and antimicrobial susceptibility of bacteria after cataract surgery in dogs. ANIMALS 16 client-owned dogs. PROCEDURES Samples for aerobic and anaerobic culture were obtained from the conjunctival fornices of both eyes of dogs 24 hours before (week 0) and 1, 3, and 6 weeks after cataract surgery. Topical administration of ofloxacin (every 6 hours) was initiated 12 hours before surgery and continued for 3 weeks. In vitro antimicrobial susceptibility was determined by Kirby-Bauer disk diffusion and a commercially available test for ofloxacin. RESULTS Frequency of positive culture results was significantly higher at week 6 than at weeks 0 and 1. Bacterial load was more likely to be moderate or high at weeks 3 and 6 than at weeks 0 and 1. The most frequently cultured organism was Staphylococcus pseudintermedius (21/78 [26.9%]), followed by coagulase-negative Staphylococcus spp (19/78 [24.4%]). Staphylococcus pseudintermedius was the organism most frequently cultured at weeks 0 (5/12), 1 (4/12), and 6 (8/19), whereas frequency of this organism was lowest at week 3 (1/20). In contrast, coagulase-negative Staphylococcus spp were the most frequently cultured organisms at week 3 (10/20). There was a significant increase in the proportion of organisms resistant to ofloxacin at week 3, compared with the proportion at week 0. CONCLUSIONS AND CLINICAL RELEVANCE The number of bacterial organisms increased and the population of conjunctival bacteria was altered and had a higher proportion resistant to ofloxacin during the 6 weeks after cataract surgery for dogs treated by use of this protocol.
Показать больше [+] Меньше [-]Immunologic responses in corn snakes (Pantherophis guttatus) after experimentally induced infection with ferlaviruses Полный текст
2017
OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses. ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes. PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days −6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed. RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses. CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.
Показать больше [+] Меньше [-]Biomechanical comparison of a 3.5-mm conical coupling plating system and a 3.5-mm locking compression plate applied as plate-rod constructs to an experimentally created fracture gap in femurs of canine cadavers Полный текст
2017
Tremolada, Giovanni | Lewis, Daniel D. | Pragnani, Ken Luka | Conrad, Bryan P. | Kim, Stanley E. | Pozzi, Antonio
OBJECTIVE To compare stiffness and resistance to cyclic fatigue of two 3.5-mm locking system plate-rod constructs applied to an experimentally created fracture gap in femurs of canine cadavers. SAMPLE 20 femurs from cadavers of 10 mixed-breed adult dogs. PROCEDURES 1 femur from each cadaver was stabilized with a conical coupling plating system-rod construct, and the contralateral femur was stabilized with a locking compression plate (LCP)-rod construct. An intramedullary Steinmann pin was inserted in each femur. A 40-mm gap then was created; the gap was centered beneath the central portion of each plate. Cyclic axial loading with increasing loads was performed. Specimens that did not fail during cyclic loading were subjected to an acute load to failure. RESULTS During cyclic loading, significantly more LCP constructs failed (6/10), compared with the number of conical coupling plating system constructs that failed (1/10). Mode of failure of the constructs included fracture of the medial or caudal aspect of the cortex of the proximal segment with bending of the plate and pin, bending of the plate and pin without fracture, and screw pullout. Mean stiffness, yield load, and load to failure were not significantly different between the 2 methods of stabilization. CONCLUSIONS AND CLINICAL RELEVANCE Both constructs had similar biomechanical properties, but the conical coupling plating system was less likely to fail than was the LCP system when subjected to cyclic loading. These results should be interpreted with caution because testing was limited to a single loading mode.
Показать больше [+] Меньше [-]Pharmacokinetics of cefazolin for prophylactic administration to dogs Полный текст
2017
Gonzalez, Omar J. | Renberg, Walter C. | Roush, James K. | Kukanich, Butch | Warner, Matt
OBJECTIVE To evaluate pharmacokinetics of cefazolin after IV injection of cefazolin (22 mg/kg) and after simultaneous IV and IM injections of cefazolin (total dose, 44 mg/kg) to dogs. ANIMALS 12 adult Beagles. PROCEDURES Dogs (6/group) were assigned to receive a single injection of cefazolin (IV group; 22 mg/kg, IV) or simultaneous injections (IV + IM group; 22 mg/kg, IV, and 22 mg/kg, IM). Interstitial fluid was collected over a 5-hour period by use of ultrafiltration probes for pharmacokinetic analysis. RESULTS Mean cefazolin concentration in the interstitial fluid at 1, 1.5, 2, 3, 4, and 5 hours after injection was 39.6, 29.1, 21.2, 10.3, 6.4, and 2.7 μg/mL, respectively, for the IV group and 38.3, 53.3, 46.4, 31.7, 19.1, and 8.9 μg/mL, respectively, for the IV + IM group. Mean area under the concentration-time curve extrapolated to infinity, maximum concentration, half-life, and time to maximum concentration was 74.99 and 154.16 h·μg/mL, 37.3 and 51.5 μg/mL, 0.96 and 1.11 hours, and 1.28 and 1.65 hours, respectively, for the IV and IV + IM groups. CONCLUSIONS AND CLINICAL RELEVANCE Cefazolin concentrations in interstitial fluid of dogs were maintained at > 4 μg/mL for 4 hours after a single IV injection and for 5 hours after simultaneous IV and IM injections. Therefore, simultaneous IV and IM administration of cefazolin 30 to 60 minutes before surgery should provide interstitial fluid concentrations effective against the most common commensal organisms (Staphylococcus spp and Streptococcus spp) on the skin of dogs for surgical procedures lasting ≤ 4 hours.
Показать больше [+] Меньше [-]Body weight, blood pressure, and systemic changes following low-dosage prednisolone administration in dogs Полный текст
2017
Yi, Sang-gwŏn | Park, Seungjo | Cheon, Byunggyu | Kim, Hyun Woo | Yu, Dohyeon | Choi, Jihye
OBJECTIVE To investigate systemic changes following low-dosage prednisolone administration in dogs. ANIMALS 4 healthy purpose-bred adult male Beagles. PROCEDURES Dogs were administered prednisolone PO at a dosage of 2 mg/kg/d for 2 weeks, 1 mg/kg/d for 4 weeks, and 0.5 mg/kg/d for 3 weeks. Body weight, blood pressure, hepatic size and echogenicity, percentage of vacuolated hepatocytes, serum hepatic enzyme activities and glucose concentration, adrenal gland size, and pancreatic echogenicity were evaluated weekly for 9 weeks. RESULTS The only significant change identified was an increase in hepatic echogenicity, assessed by measuring liver-kidney contrast on ultrasonographic images. Increases in hepatic size and percentage of vacuolated hepatocytes were identified, but values did not differ from baseline values. Similarly, serum hepatic enzyme activities increased, but changes were mild and not significantly different from baseline values. Body weight, pancreatic echogenicity, and serum glucose concentration did not show noticeable changes. Mild systemic hypertension was seen, but blood pressure was not significantly different from the baseline value. Similarly, adrenal gland size steadily decreased during the first 6 weeks and increased again after the prednisolone dosage was decreased to 0.5 mg/kg/d. However, mean adrenal gland size was not significantly different from the baseline value at any time. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that in dogs, administration of prednisolone at a low dosage was associated with minimal systemic effects.
Показать больше [+] Меньше [-]Bile acid composition of gallbladder contents in dogs with gallbladder mucocele and biliary sludge Полный текст
2017
Kakimoto, Toshiaki | Kanemoto, Hideyuki | Fukushima, Kenjiro | Ohno, Koichi | Tsujimoto, Hajime
OBJECTIVE To examine bile acid composition of gallbladder contents in dogs with gallbladder mucocele and biliary sludge. ANIMALS 18 dogs with gallbladder mucocele (GBM group), 8 dogs with immobile biliary sludge (i-BS group), 17 dogs with mobile biliary sludge (m-BS group), and 14 healthy dogs (control group). PROCEDURES Samples of gallbladder contents were obtained by use of percutaneous ultrasound-guided cholecystocentesis or during cholecystectomy or necropsy. Concentrations of 15 bile acids were determined by use of highperformance liquid chromatography, and a bile acid compositional ratio was calculated for each group. RESULTS Concentrations of most bile acids in the GBM group were significantly lower than those in the control and m-BS groups. Compositional ratio of taurodeoxycholic acid, which is 1 of 3 major bile acids in dogs, was significantly lower in the GBM and i-BS groups, compared with ratios for the control and m-BS groups. The compositional ratio of taurocholic acid was significantly higher and that of taurochenodeoxycholic acid significantly lower in the i-BS group than in the control group. CONCLUSIONS AND CLINICAL RELEVANCE In this study, concentrations and fractions of bile acids in gallbladder contents were significantly different in dogs with gallbladder mucocele or immobile biliary sludge, compared with results for healthy control dogs. Studies are needed to determine whether changes in bile acid composition are primary or secondary events of gallbladder abnormalities.
Показать больше [+] Меньше [-]Effects of a combination of acepromazine maleate and butorphanol tartrate on conventional and two-dimensional speckle tracking echocardiography in healthy dogs Полный текст
2017
Santraelli, Giorgia | Lopez, Jesus Talavera | Palacio, Josefa Ferandez Del
OBJECTIVE To determine effects of a combination of acepromazine maleate and butorphanol tartrate on conventional echocardiographic variables and on strain values obtained by use of 2-D speckle tracking echocardiography (STE) in healthy dogs. ANIMALS 18 healthy medium- and large-size adult dogs. PROCEDURES Transthoracic echocardiographic examination (2-D, M-mode, color flow, spectral Doppler, and tissue Doppler ultrasonography) and high-definition oscillometric blood pressure measurement were performed before and after dogs were sedated by IM administration of a combination of acepromazine (0.02 mg/kg) and butorphanol (0.2 mg/kg). Adequacy of sedation for echocardiographic examination was evaluated. Circumferential and longitudinal global and segmental strains of the left ventricle (LV) were obtained with 2-D STE by use of right parasternal short-axis and left parasternal apical views. Values before and after sedation were compared. RESULTS The sedation combination provided adequate immobilization to facilitate echocardiographic examination. Heart rate and mean and diastolic blood pressures decreased significantly after dogs were sedated. A few conventional echocardiographic variables differed significantly from baseline values after sedation, including decreased end-diastolic LV volume index, peak velocity of late diastolic transmitral flow, and late diastolic septal mitral and tricuspid annulus velocities, increased ejection time, and increased mitral ratio of peak early to late diastolic filling velocity; global strain values were not affected, but 1 segmental (apical lateral) strain value decreased significantly. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that acepromazine and butorphanol at the doses used in this study provided sedation adequate to facilitate echocardiography, with only mild influences on conventional and 2-D STE variables.
Показать больше [+] Меньше [-]Seroprevalence of Mycoplasma suis infection in pigs in eastern China as estimated by a blocking enzyme-linked immunosorbent assay Полный текст
2017
Zhongyang, Liang | Jiansong, Zhang | Yijuan, Shen | Yuting, Xia | Yufeng, Li | Jiarong, Xu
Porcine infectious anemia caused by Mycoplasma suis is a global disease and results in serious economic losses. To determine the prevalence of M. suis infection in eastern China, a cross-sectional serologic study was conducted with 3458 porcine serum samples randomly obtained from January 2014 to August 2016. The samples were tested with a blocking enzyme-linked immunosorbent assay developed in our laboratory. The seroprevalence was 33.3% in the complete sample set and was 25.9%, 37.8%, and 37.8% in 2014, 2015, and 2016, respectively. The seroprevalence was distinctly higher in summer (39.9%) and autumn (42.0%) than in spring (28.9%) and winter (23.3%). Shanghai was the region with the highest seroprevalence (54.2%) and Jiangsu the region with the lowest (23.0%). The seroprevalence was markedly higher in boars (47.1%), multiparous sows (47.0%), and replacement gilts (39.2%) than in piglets (24.2%), fattening pigs (17.2%), and nursery pigs (12.5%). These data demonstrate that the prevalence of M. suis infection is increasing yearly in eastern China.
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