Dynamics of African horse sickness virus nucleic acid and antibody in horses following immunization with a commercial polyvalent live attenuated vaccine.

African horse sickness (AHS) is a fatal disease of equids relevant to the global equine industry. Detection of AHS virus (AHSV) during outbreaks has become more rapid and efficient with the advent of group specific reverse transcriptase quantitative polymerase chain reaction (GS RT-qPCR) assays to detect AHSV nucleic acid. Use of GS RT-qPCR together with recently described type specific (TS RT-qPCR) assays cannot only expedite diagnosis of AHS but also facilitate further evaluation of the dynamics of AHSV infection in the equine host. A potential limitation to the application of these assays is that they detect viral nucleic acid originating from any AHS live attenuated vaccine (LAV), which is the vaccine type routinely administered to horses in South Africa. The aim of this study was to contrast the dynamics and duration of the RNAaemia to the serological responses of horses following immunization with a commercial polyvalent AHSV-LAV using GS and TS RT-qPCR assays and serum neutralisation tests. The results of the study showed extended RNAemia in vaccinated horses, and that more horses tested positive on GS RT-qPCR with lower Cq values after receiving the AHSV-LAV containing types 1, 3 and 4 prior to the vaccine containing types 2, 6, 7 and 8, rather than when the vaccine combinations were reversed. Furthermore, lower Cq values were obtained when vaccines were administered 4weeks apart as compared with a longer interval or 12weeks apart. These findings are of particular relevance in regions where AHSV-LAVs are used as the use of these vaccines may complicate the accurate interpretation of diagnostic testing results.

Electromechanical abdominal massage and colonic function in individuals with a spinal cord injury and chronic bowel problems.

STUDY DESIGN: A prospective intervention of noninvasive abdominal massage using an electromechanical apparatus on bowel function in individuals with spinal cord injury (SCI). OBJECTIVES: To evaluate the effects of noninvasive abdominal massage using an electromechanical apparatus on bowel function in individuals with SCI and chronic bowel problems. This easy-to-use apparatus can be applied by the patients at home without the help of a therapist. SETTING: Homes of community-living individuals. METHODS: Twenty-one subjects with SCI were instructed to use the massage apparatus daily for 20 min during a 10-week period. Compliance, effects, side effects and user satisfaction were assessed using questionnaires. RESULTS: Fifteen subjects completed the 10-week period. Although some characteristics of defecation changed positively for some of the subjects (time to result, amount, consistency), none felt better or more confident after using the massage device. In addition, some individuals experienced negative side effects (predominantly pain or discomfort). The overall satisfaction with the device is ambiguous, with half of the group judging the device as insufficient and the other half as at least adequate. CONCLUSION: The use of an electromechanical massage device does not improve bowel function in most individuals with SCI who have chronic bowel problems. Why some subjects benefit and others do not should be investigated in future studies.

Serological response of foals to polyvalent and monovalent live-attenuated African horse sickness virus vaccines.

African horse sickness (AHS) is typically a highly fatal disease in susceptible horses and vaccination is currently used to prevent the occurrence of disease in endemic areas. Similarly, vaccination has been central to the control of incursions of African horse sickness virus (AHSV) into previously unaffected areas and will likely play a significant role in any future incursions. Horses in the AHSV-infected area in South Africa are vaccinated annually with a live-attenuated (modified-live virus [MLV]) vaccine, which includes a cocktail of serotypes 1, 3, 4 (bottle 1) and 2, 6-8 (bottle 2) delivered in two separate doses at least 21 days apart. In this study, the neutralising antibody response of foals immunized with this polyvalent MLV AHSV vaccine was evaluated and compared to the response elicited to monovalent MLV AHSV serotypes. Naïve foals were immunized with either the polyvalent MLV AHSV vaccine, or a combination of monovalent MLV vaccines containing individual AHSV serotypes 1, 4, 7 or 8. There was a marked and consistent difference in the immunogenicity of individual virus serotypes contained in the MLV vaccines. Specifically, foals most consistently seroconverted to AHSV-1 and responses to other serotypes were highly variable, and often weak or not detected. The serotype-specific responses of foals given the monovalent MLV vaccines were similar to those of foals given the polyvalent MLV preparation suggesting that there is no obvious enhanced immune response through the administration of a monovalent vaccine as opposed to the polyvalent vaccine.

Development of a real time polymerase chain reaction assay for equine encephalosis virus.

Equine encephalosis virus (EEV) is the cause of equine encephalosis. The disease is similar to mild forms of African horse sickness (AHS) and the two diseases are easily confused. Laboratory identification and serotyping of EEV is based on viral isolation in BHK-21 cells and a viral plaque inhibition neutralisation test. These procedures are time-consuming and therefore a more rapid diagnostic assay for EEV that can distinguish EEV from African horse sickness virus (AHSV) infections was developed. The S7 (VP7) gene from 38 EEV isolates representing all seven serotypes was amplified and sequenced. A conserved region at the 5' end of the gene was identified and used to design group-specific EEV primers and a TaqMan(®) MGB™ hydrolysis probe. The efficiency of the EEV real-time RT-PCR assay was 81%. The assay was specific, as it did not detect any of the nine serotypes of AHSV, nor 24 serotypes of bluetongue virus (BTV) and sensitive, with a 95% limit of detection of 10(2.9) TCID50/ml blood (95% confidence interval: 10(2.7) to 10(3.3)). The real-time format was selected because of its convenience, sensitivity and ability to produce results rapidly.

Passive transfer and rate of decay of maternal antibody against African horse sickness virus in South African Thoroughbred foals.

REASONS FOR PERFORMING STUDY: African horse sickness is an insect-transmitted, noncontagious disease of equids caused by African horse sickness virus (AHSV). Mortality can exceed 90% in fully susceptible horse populations. A live-attenuated (modified live) cell-culture-adapted (MLV) polyvalent AHSV vaccine is widely used to control African horse sickness in endemic areas in southern Africa. Field studies detailing antibody responses of vaccinated horses are lacking. OBJECTIVES: To determine antibody titres to the 9 known serotypes of AHSV in a cohort of broodmares that were regularly vaccinated with the MLV AHSV vaccine and to measure the passive transfer and rate of decay of maternal antibody to the individual virus serotypes in foals. METHODS: Serum was collected from 15 mares before foaling and from their foals after foaling and monthly thereafter for 6 months. Antibody titres to each of the 9 AHSV serotypes were determined by serum virus neutralisation assay. RESULTS: There was marked variation in the antibody response of the mares to individual AHSV serotypes even after repeated vaccination, with consistently higher titre responses to some virus serotypes. Likewise, the duration of maternally derived antibodies in foals differed among serotypes. CONCLUSIONS: Data from this study confirm variation of the neutralising antibody response of individual mares to repeated vaccination with polyvalent AHSV vaccine. Virus strains of individual AHSV serotypes included in the vaccine may vary in their inherent immunogenicity. Passively acquired maternal antibodies to AHSV vary markedly among foals born to vaccinated mares, with further variation in the duration of passive immunity to individual AHSV serotypes. POTENTIAL RELEVANCE: These data are relevant to the effective utilisation of live-attenuated AHSV vaccines in endemic regions, and potentially to the use of vaccines in response to future incursions of AHSV into previously free regions. Further studies involving a larger population will be required to determine the optimal time for vaccinating foals.

African horse sickness in naturally infected, immunised horses.

To determine whether subclinical cases, together with clinical cases, of African horse sickness (AHS) occur in immunised horses in field conditions, whole blood samples were collected and rectal temperatures recorded weekly from 50 Nooitgedacht ponies resident in open camps at the Faculty of Veterinary Science, University of Pretoria, Onderstepoort, during 2008-2010. The samples were tested for the presence of African horse sickness virus (AHSV) RNA by a recently developed real-time RT-PCR. It was shown that 16% of immunised horses in an AHS endemic area were infected with AHSV over a 2 year period, with half of these (8%) being subclinically infected. The potential impact of such cases on the epidemiology of AHS warrants further investigation.

Development and optimisation of a duplex real-time reverse transcription quantitative PCR assay targeting the VP7 and NS2 genes of African horse sickness virus.

Nucleotide sequences of 52 South African isolates of African horse sickness virus (AHSV) collected during 2004-2005 and including viruses of all nine AHSV serotypes, were used to design and develop a duplex real-time reverse transcription quantitative PCR (RT-PCR) assay targeting the VP7 (S8) and NS2 (S9) genes of AHSV. The assay was optimized for detection of AHSV in fresh and frozen blood of naturally infected horses. Assay performance was enhanced using random hexamers rather than gene-specific primers for RT, and with denaturation of double-stranded RNA in the presence of random hexamers. The assay was efficient with a linear range of at least five orders of magnitude. The analytical sensitivity of the assay was 132 copies of the target genes (4125 copies per ml of blood), and the assay was at least 10-fold more sensitive than virus isolation on BHK-21 cells. The assay was also highly specific because it did not detect related orbiviruses, such as bluetongue and equine encephalosis viruses.

Pressure ulcer care: nutritional therapy need not add to costs.

Fewer patients with pressure ulcers in Dutch nursing homes receive nutritional therapy via sip feeds, possibly because of cost concerns. But this therapy would not cost more if it reduced the duration of nursing care by even one day, this paper argues.

Blood chemical and electrolyte concentrations in the mountain reedbuck Redunca fulvorufula.

Concentrations of sodium, potassium, chloride, inorganic phosphorus, total magnesium, total calcium, iron, urea, creatinine, total protein, albumin, total bilirubin, alkaline phosphatase (ALP), alanine transaminase (ALT), lactate dehydrogenase (LD), creatine kinase (CK), gamma-glutamyltransferase (GGT) and aspartate transaminase (AST) were determined in serum specimens collected from 53 free-ranging mountain reedbuck (Redunca fulvorufula) during live capture using nets. Considerable variations in the concentrations of the enzymes ALP, LDH, CK, GGT and AST were found as well as in the concentrations of creatinine, bilirubin and iron. This wide variation in results seriously questions the usefulness of similar blood investigations on heterogenous groups of mechanically restrained animals.