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Evaluation of plant-produced Clostridium perfringens type D epsilon toxoid in a vaccine against enterotoxaemia in sheep Full text
2017
Mokoena, Tinyiko(Council for Scientific and Industrial Research ,University of Pretoria Forestry and Agricultural Biotechnology Institute Department of Plant Science) | Chakauya, Ereck(Council for Scientific and Industrial Research) | Crampton, Michael(Council for Scientific and Industrial Research) | Weyers, Boet(Onderstepoort Biological Products) | Tselanyane, Malefa(Council for Scientific and Industrial Research) | Tsekoa, Tsepo(Council for Scientific and Industrial Research) | Chikwamba, Rachel(Council for Scientific and Industrial Research)
Evaluation of plant-produced Clostridium perfringens type D epsilon toxoid in a vaccine against enterotoxaemia in sheep Full text
2017
Mokoena, Tinyiko(Council for Scientific and Industrial Research ,University of Pretoria Forestry and Agricultural Biotechnology Institute Department of Plant Science) | Chakauya, Ereck(Council for Scientific and Industrial Research) | Crampton, Michael(Council for Scientific and Industrial Research) | Weyers, Boet(Onderstepoort Biological Products) | Tselanyane, Malefa(Council for Scientific and Industrial Research) | Tsekoa, Tsepo(Council for Scientific and Industrial Research) | Chikwamba, Rachel(Council for Scientific and Industrial Research)
Enterotoxaemia (pulpy kidney) is a common bacterial disease of sheep caused by Clostridium perfringens type D epsilon toxin. It has mortality rates of up to 30% in non-vaccinated animals. Current vaccines from whole cell cultures are expensive to manufacture and can induce local inflammatory responses in sheep. They usually have reduced immunogenicity because of the difficulty of standardising the inactivation step in vaccine manufacturing. In the current study, we evaluated the safety and potency of a recombinant plant-made epsilon toxoid protein (r-Etox) as an affordable and safer alternative vaccine for developing countries. Results of injection site reactions, rectal temperature and toxin neutralisation test in single and prime-boost inoculations of mice, guinea pigs and sheep suggest that the product is not toxic to animals and could protect sheep against enterotoxaemia.
Show more [+] Less [-]The sero-prevalence and sero-incidence of African horse sickness and equine encephalosis in selected horse and donkey populations in Zimbabwe Full text
2017
Gordon, Stuart J.G.(Massey University Institute of Veterinary, Animal and Biomedical Sciences) | Bolwell, Charlotte(Massey University Institute of Veterinary, Animal and Biomedical Sciences) | Rogers, Chris W.(Massey University Institute of Veterinary, Animal and Biomedical Sciences) | Musuka, Godfrey(Columbia University Mailman School of Public Health International Center for AIDS Care and Treatment Programs) | Kelly, Patrick(Ross University School of Veterinary Medicine) | Guthrie, Alan(University of Pretoria Faculty of Veterinary Science Equine Research Centre) | Mellor, Philip S.(The Pirbright Institute International Research Centre) | Hamblin, Christopher(The Pirbright Institute International Research Centre)
The sero-prevalence and sero-incidence of African horse sickness and equine encephalosis in selected horse and donkey populations in Zimbabwe Full text
2017
Gordon, Stuart J.G.(Massey University Institute of Veterinary, Animal and Biomedical Sciences) | Bolwell, Charlotte(Massey University Institute of Veterinary, Animal and Biomedical Sciences) | Rogers, Chris W.(Massey University Institute of Veterinary, Animal and Biomedical Sciences) | Musuka, Godfrey(Columbia University Mailman School of Public Health International Center for AIDS Care and Treatment Programs) | Kelly, Patrick(Ross University School of Veterinary Medicine) | Guthrie, Alan(University of Pretoria Faculty of Veterinary Science Equine Research Centre) | Mellor, Philip S.(The Pirbright Institute International Research Centre) | Hamblin, Christopher(The Pirbright Institute International Research Centre)
Sentinel herds and samples submitted by private equine practitioners were used to determine the sero-prevalence and sero-incidence of African horse sickness virus (AHSV) and equine encephalosis virus (EEV) in horse and donkey populations in the Highveld region of Zimbabwe. The sero-prevalence and sero-incidence of antibodies against these viruses were determined using the competitive enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibodies. In donkeys, the median sero-prevalence of AHSV antibodies, across the three rainy seasons under study, was 75% (inter quartile range [IQR] 67-83), with a seasonal median sero-incidence of 45% (IQR 40-63). In horses, the median sero-prevalence of EEV antibodies was 63% (IQR 21-73), with a median seasonal sero-incidence of 10.5% (IQR 10-14), while in donkeys the median sero-prevalence of EEV antibodies was 80% (IQR 67-90), with a median seasonal sero-incidence of 50% (IQR 40-60). This study highlighted the significant levels of exposure of donkeys to AHSV and horses and donkeys to EEV in Zimbabwe despite equine encephalosis remaining unreported by Zimbabwean veterinarians to date. Most seroconversions in sentinel herd animals to AHSV and EEV occurred towards the end of the rainy season in March, April and May corresponding to the time of the year when the Culicoides vectors are in high abundance. In order to determine the clinical significance of these infections, blood and spleen samples, submitted by private equine veterinary practitioners over a 5-year period, from horses showing characteristic clinical signs of African horse sickness were tested for the presence of viral antigen using the antigen capture ELISA. The median sero-prevalence of AHSV antigen in horses recorded from these samples was 38% (IQR 33-88). The predominant AHSV antigen from these samples was serotype 7 (33%) followed by serotype 2 (26%) and serotypes 4 and 8 (16% each). African horse sickness virus serotypes 3 and 9, identified in this study, had not been previously reported in Zimbabwe.
Show more [+] Less [-]The sero-prevalence and sero-incidence of African horse sickness and equine encephalosis in selected horse and donkey populations in Zimbabwe Full text
2017
Stuart J.G. Gordon | Charlotte Bolwell | Chris W. Rogers | Godfrey Musuka | Patrick Kelly | Alan Guthrie | Philip S. Mellor | Christopher Hamblin
Sentinel herds and samples submitted by private equine practitioners were used to determine the sero-prevalence and sero-incidence of African horse sickness virus (AHSV) and equine encephalosis virus (EEV) in horse and donkey populations in the Highveld region of Zimbabwe. The sero-prevalence and sero-incidence of antibodies against these viruses were determined using the competitive enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibodies. In donkeys, the median sero-prevalence of AHSV antibodies, across the three rainy seasons under study, was 75% (inter quartile range [IQR] 67–83), with a seasonal median sero-incidence of 45% (IQR 40–63). In horses, the median sero-prevalence of EEV antibodies was 63% (IQR 21–73), with a median seasonal sero-incidence of 10.5% (IQR 10–14), while in donkeys the median sero-prevalence of EEV antibodies was 80% (IQR 67–90), with a median seasonal sero-incidence of 50% (IQR 40–60). This study highlighted the significant levels of exposure of donkeys to AHSV and horses and donkeys to EEV in Zimbabwe despite equine encephalosis remaining unreported by Zimbabwean veterinarians to date. Most seroconversions in sentinel herd animals to AHSV and EEV occurred towards the end of the rainy season in March, April and May corresponding to the time of the year when the Culicoides vectors are in high abundance. In order to determine the clinical significance of these infections, blood and spleen samples, submitted by private equine veterinary practitioners over a 5-year period, from horses showing characteristic clinical signs of African horse sickness were tested for the presence of viral antigen using the antigen capture ELISA. The median sero-prevalence of AHSV antigen in horses recorded from these samples was 38% (IQR 33–88). The predominant AHSV antigen from these samples was serotype 7 (33%) followed by serotype 2 (26%) and serotypes 4 and 8 (16% each). African horse sickness virus serotypes 3 and 9, identified in this study, had not been previously reported in Zimbabwe.
Show more [+] Less [-]Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12) Full text
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) Full text
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.
Show more [+] Less [-]Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12) Full text
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.
Show more [+] Less [-]Assessment of a portable lactate meter for field use in the white rhinoceros (Ceratotherium simum) Full text
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) Full text
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.
Show more [+] Less [-]Assessment of a portable lactate meter for field use in the white rhinoceros (<i>Ceratotherium simum</i>) Full text
2017
Georgina C. Cole | Adrian S.W. Tordiffe | Gerhard Steenkamp
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.
Show more [+] Less [-]Alterations in conjunctival bacteria and antimicrobial susceptibility during topical administration of ofloxacin after cataract surgery in dogs Full text
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.
Show more [+] Less [-]Immunologic responses in corn snakes (Pantherophis guttatus) after experimentally induced infection with ferlaviruses Full text
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.
Show more [+] Less [-]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 Full text
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.
Show more [+] Less [-]Pharmacokinetics of cefazolin for prophylactic administration to dogs Full text
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.
Show more [+] Less [-]Body weight, blood pressure, and systemic changes following low-dosage prednisolone administration in dogs Full text
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.
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