<strong>How to cite this article:</strong> Geysen, D. &amp; Berkvens, D., 2013, ‘Lack of evidence for safe vaccination with the Muguga cocktail in Sudan’, <em>Onderstepoort Journal of Veterinary Research</em> 80(1), Art. #571, 1 page. http:// dx.doi.org/10.4102/ojvr. v80i1.571

Objective: To evaluate agents used for delivery of small interfering RNAs (siRNAs) into feline corneal cells, toxicity of the delivery agents, and functionality of anti-feline herpesvirus 1 (FHV-1)–specific siRNA combinations. Sample: Feline primary corneal cells and 19 six-month-old colony-bred cats. Procedures: siRNA delivery into corneal cells via various delivery agents was evaluated via flow cytometric detection of labeled siRNAs. Cellular toxicity was evaluated with a proliferation assay. Functionality was tested via quantitative reverse transcriptase PCR assay, plaque assay, and flow cytometry. In vivo safety was evaluated with an ocular scoring method following topical application of delivery agents containing siRNAs into eyes. Corneal biopsy specimens were used to assess safety and uptake of siRNAs into corneal cells. Results: Use of 3 delivery agents resulted in > 95% transfection of primary corneal cells. Use of a peptide for ocular delivery yielded approximately 82% transfection of cells in vitro. In cultured corneal cells, use of the siRNA combinations resulted in approximately 76% to 89% reduction in FHV-1–specific mRNA, 63% to 67% reduction of FHV-1–specific proteins in treated cells, and 97% to 98% reduction in FHV-1 replication. The agents were nonirritating in eyes, caused no substantial clinical ocular signs, and were nontoxic. Histologically, corneal epithelium and stroma were normal in treated cats. However, none of the agents were effective in delivering siRNAs into the corneal cells in vivo. Conclusions and Clinical Relevance: The tested anti–FHV-1–specific siRNAs could potentially be used as a treatment for FHV-1 if a successful means of in vivo delivery can be achieved.

Objective: To develop a broad-range 28S ribosomal DNA quantitative PCR (qPCR) assay for detection of fungal DNA in equine endometrial samples. Sample: 12 fungal samples from a clinical diagnostic laboratory and 29 samples obtained from 17 mares. Procedures: The qPCR assay was optimized with commercially acquired fungal organisms and validated with samples obtained from the clinical diagnostic laboratory. Subsequently, 29 samples from 17 mares suspected of having fungal endometritis were evaluated via the qPCR assay and via traditional fungal culture and endometrial cytology. Amplicons from the qPCR assay were subjected to genetic sequencing to identify the organisms. Results: The qPCR assay theoretically had a detection threshold of 2 organisms of Candida albicans. Fungal DNA was amplified from all 12 fungal samples from the commercial diagnostic laboratory. Fungal identification by use of genetic sequencing was successful for 34 of 36 amplicons from the 12 samples assayed. A fungal agent was identified via qPCR assay and genetic sequencing in all 12 samples; in contrast, a fungal agent was identified in only 8 of 12 samples via standard fungal culture and biochemical analysis. The qPCR assay detected fungal DNA in samples from 12 of 17 mares suspected of having fungal endometritis. Conclusions and Clinical Relevance: A rapid, sensitive, and repeatable qPCR assay was developed for detection of fungal DNA from equine endometrial samples. The qPCR may prove to be clinically useful as an adjunct to microbial culture and cytologic examination to provide identification of fungal organisms in a timely manner.

Objective: To identify proteins with differential expression between healthy dogs and dogs with stifle joint osteoarthritis secondary to cranial cruciate ligament (CCL) disease. Sample: Serum and synovial fluid samples obtained from dogs with stifle joint osteoarthritis before (n = 10) and after (8) surgery and control dogs without osteoarthritis (9) and archived synovial membrane and articular cartilage samples obtained from dogs with stifle joint osteoarthritis (5) and dogs without arthritis (5). Procedures: Serum and synovial fluid samples were analyzed via liquid chromatography–tandem mass spectrometry; results were compared against a nonredundant protein database. Expression of complement component 3 in archived tissue samples was determined via immunohistochemical methods. Results: No proteins had significantly different expression between serum samples of control dogs versus those of dogs with stifle joint osteoarthritis. Eleven proteins (complement component 3 precursor, complement factor I precursor, apolipoprotein B-100 precursor, serum paraoxonase and arylesterase 1, zinc-alpha-2-glycoprotein precursor, serum amyloid A, transthyretin precursor, retinol-binding protein 4 precursor, alpha-2-macroglobulin precursor, angiotensinogen precursor, and fibronectin 1 isoform 1 preproprotein) had significantly different expression (> 2.0-fold) between synovial fluid samples obtained before surgery from dogs with stifle joint osteoarthritis versus those obtained from control dogs. Complement component 3 was strongly expressed in all (5/5) synovial membrane samples of dogs with stifle joint osteoarthritis and weakly expressed in 3 of 5 synovial membrane samples of dogs without stifle joint arthritis. Conclusions and Clinical Relevance: Findings suggested that the complement system and proteins involved in lipid and cholesterol metabolism may have a role in stifle joint osteoarthritis, CCL disease, or both.

Objective: To assess vascular changes induced by hyperadrenocorticism of hyperplastic adrenal glands in dogs via contrast-enhanced ultrasonography. Animals: 12 dogs with pituitary-dependent hyperadrenocorticism (PDH) and 7 healthy control dogs ≥ 7 years old. Procedures: Dogs were assigned to the PDH and control groups and to small-breed (n = 6), medium-breed (4), and large-breed (9) subgroups. Contrast-enhanced ultrasonography of both adrenal glands in each dog was performed with IV injections of contrast agent. Time-intensity curves for the adrenal cortex, adrenal medulla, and ipsilateral renal artery of both adrenal glands were generated. Perfusion variables (time to peak [TTP], upslope of wash-in phase, and downslope of washout phase) were calculated. Results: Contrast-enhanced ultrasonography revealed no qualitative difference between PDH and control groups. Quantitatively, TTPs were longer in the adrenal cortex and adrenal medulla of the PDH group, compared with values for the control group, particularly in the adrenal cortex and adrenal medulla of the small-breed subgroup. Washout downslopes were lower for the renal artery, adrenal cortex, and adrenal medulla of the small-breed subgroup between the PDH and control groups. No other perfusion variables differed between groups. Conclusions and Clinical Relevance: Contrast-enhanced ultrasonography of the adrenal glands in dogs with PDH revealed a delayed TTP in the adrenal cortex and adrenal medulla, compared with values for control dogs. Contrast-enhanced ultrasonography was able to detect vascular changes induced by hyperadrenocorticism. Further studies are needed to evaluate whether reference ranges for clinically normal dogs and dogs with PDH can be determined and applied in clinical settings.

Objective: To evaluate associations between economic and performance outcomes with the number of treatments after an initial diagnosis of bovine respiratory disease (BRD) in commercial feedlot cattle. Animals: 212,867 cattle arriving in a Midwestern feedlot between 2001 and 2006. Procedures: An economic model was created to estimate net returns. Generalized linear mixed models were used to determine associations between the frequency of BRD treatments and other demographic variables with economic and performance outcomes. Results: Net returns decreased with increasing number of treatments for BRD. However, the magnitude depended on the season during which cattle arrived at the feedlot, with significantly higher returns for cattle arriving during fall and summer than for cattle arriving during winter and spring. For fall arrivals, there were higher mean net returns for cattle that were never treated ($39.41) than for cattle treated once ($29.49), twice ($16.56), or ≥ 3 times (−$33.00). For summer arrivals, there were higher least squares mean net returns for cattle that were never treated ($31.83) than for cattle treated once ($20.22), twice ($6.37), or ≥ 3 times ($−42.56). Carcass traits pertaining to weight and quality grade were deemed responsible for differences in net returns among cattle receiving different numbers of treatments after an initial diagnosis of BRD. Conclusions and Clinical Relevance: Differences in economic net returns and performance outcomes for feedlot cattle were determined on the basis of number of treatments after an initial diagnosis of BRD; the analysis accounted for the season of arrival, sex, and weight class.

A total of 257 fishes from four families, Clariidae, Cichlidae, Cyprinidae and Schilbeidae were collected from three localities: the Sand River Dam, Swaziland; the Nylsvlei Nature Reserve, South Africa and the Vaal Dam and Vaal River Barrage, South Africa. Only fishes (n= 154) from Clariidae and Cichlidae were found to be infected with trypanosomes. A total of 221 Clarias gariepinus (Burchell 1822) were collected from the Vaal Dam and Vaal Barrage area, South Africa. Of these, 74%(89/121) were infected with trypanosomes from the Vaal Dam and 63%(63/100) from the Vaal River Barrage, with no seasonal infection pattern. A prevalence of 25%(1/4) was found in C. gariepinus from the Sand River Dam, Swaziland, and a 50% (1/2) prevalence was found in Tilapia sparrmanii from the Nylsvlei Nature Reserve, South Africa. Standard measurements conformed closely to the morphometric and morphological descriptions of Trypanosoma mukasai. This article provides new locality records for T. mukasai from the Vaal Dam, Vaal River Barrage and Nylsvlei Nature Reserve (South Africa) and the Sand River Dam (Swaziland). Tilapia sparrmanii collected in the Sand River Dam in Swaziland is also noted as a new host record.

Objective-To compare effects of anesthetic induction with midazolam-propofol or midazolam-etomidate on intraocular pressure (IOP), pupillary diameter (PD), pulse rate, blood pressure, and respiratory rate in clinically normal dogs. Animals-18 dogs. Procedures-Dogs undergoing ophthalmic surgery received midazolam (0.2 mg/kg, IV) and either propofol or etomidate (IV) until intubatable. For all dogs, results of physical examinations, ophthalmic examinations of the nonoperated eye, and preanesthetic blood analyses were normal. Intraocular pressure, PD, blood pressure, pulse rate, and respiratory rate were measured in the nonoperated eye at 5 time points: just prior to the anesthetic induction sequence, after 5 minutes of preanesthetic oxygenation via face mask, after IV administration of midazolam, after IV anesthetic induction, and after endotracheal intubation. Results-PD decreased significantly from baseline by 4.4 +/- 0.4 mm (mean +/- SD) after anesthetic induction and 5.3 +/- 0.4 mm after intubation in the etomidate group and by 1. 2 +/- 0.4 mm after intubation in the propofol group. Intraocular pressure was increased significantly from baseline by 3.2 +/- 1.0 mm Hg after anesthetic induction in the etomidate group and by 4.7 +/- 1.2 mm Hg after anesthetic induction and 4.5 +/- 1. 2 mm Hg after intubation in the propofol group. Pulse rate was significantly lower by 28.6 +/- 12.6 beats/min after anesthetic induction in the etomidate group, compared with the propofol group. Conclusions and Clinical Relevance-At the studied doses, midazolam-etomidate caused clinically important miosis and increased IOP. Midazolam-propofol caused an even greater increase in IOP but had minimal effects on PD.

Objective-To assess the feasibility of measuring cord dorsum potentials (CDPs) in anesthetized clinically normal dogs after caudal nerve stimulation, determine the intervertebral site of maximum amplitude and best waveform of the CDP, and evaluate the effects of neuromuscular blockade. Animals-8 male and 4 female dogs (age, 1 to 5 years). Procedures-Dogs were anesthetized, and CDPs were recorded via needles placed on the dorsal lamina at intervertebral spaces L1-2 through L7-S1. Caudal nerves were stimulated with monopolar electrodes inserted laterally to the level of the caudal vertebrae. Dogs were tested without and during neuromuscular blockade induced with atracurium besylate. The CDP latency and amplitude were determined from the largest amplitude tracings. Results-CDPs were recorded in 11 of 12 dogs without neuromuscular blockade and in all dogs during neuromuscular blockade. The CDP was largest and most isolated at the L4-5 intervertebral space (3 dogs) or the L5-6 intervertebral space (9 dogs); this site corresponded to the segment of insertion of the first caudal nerve. Onset latencies ranged from 2.0 to 4.7 milliseconds, and there was no effect of neuromuscular blockade on latencies. Amplitudes of the CDPs were highly variable for both experimental conditions. Conclusions and Clinical Relevance-CDPs were recorded from all dogs tested in the study; neuromuscular blockade was not critical for successful CDP recording but reduced muscle artifact. This technique may be useful as a tool to assess the caudal nerve roots in dogs suspected of having compressive lumbosacral disease or myelopathies affecting the lumbar intumescence.

Objective—To evaluate effects of quaternary benzo(c)phenanthridine alkaloids (QBAs) against Salmonella spp and determine effects on growth performance, organism shedding, and gastrointestinal tract integrity in pigs inoculated with Salmonella enterica serovar Typhimurium. Sample—36 Salmonella isolates and twenty 5-week-old pigs. Procedures—Minimum inhibitory concentration of QBAs against the Salmonella isolates was determined. Pigs were allocated to 4 groups and inoculated with Salmonella organisms. Pigs received diets supplemented with 1.5 g of QBAs/1,000 kg of feed, 0.75 g of QBAs/1,000 kg of feed, or 59.4 g of chlortetracycline/1,000 kg of feed or a nonsupplemented (control) diet. Pigs were weighed on day 0 and then weekly for 40 days. Fecal samples were collected to quantify Salmonella organisms. Gastrointestinal tract integrity was evaluated by measuring transepithelial resistance. Results—In vitro, 9 of 36 (25%) Salmonella isolates were inhibited at 90 μg of QBAs/mL; all 36 were inhibited at 179 μg of QBAs/mL. Diets containing QBAs significantly decreased Salmonella spp shedding; shedding was lower 40 days after inoculation for pigs fed diets containing QBAs or chlortetracycline than for pigs fed the control diet. Growth performance was similar for pigs fed diets containing QBA or chlortetracycline. Gastrointestinal tract integrity was improved in pigs fed the diet containing 1.5 g of QBAs/1,000 kg of feed. Conclusions and Clinical Relevance—QBAs and chlortetracycline decreased Salmonella spp shedding but did not differ with regard to growth performance. Gastrointestinal tract integrity was better, albeit not significantly, in pigs fed diets containing QBAs. Further investigation into the role of QBAs and their mechanism as an immunomodulator is necessary.