Prevalence of Rift Valley fever in domestic ruminants in the central and northern regions of Burkina Faso.

The seroprevalence of Rift Valley fever was determined in cattle, sheep and goats in selected areas of northern and central Burkina Faso. A total of 520 serum samples were screened for anti-Rift Valley fever virus immunoglobulin G (IgG) antibodies using an inhibition enzyme-linked immunosorbent assay (ELISA). An average seroprevalence of 7.67% (range 5% to 20%) was found in ruminants in Seno and Soum provinces, and prevalences of 20% and 22.5% in cattle in Yatenga and Oubritenga provinces, respectively. The location, species and age of the animals were found to influence the seroprevalence. All the ELISA IgG-positive samples were tested for IgM in a competitive ELISA and were found negative, thus ruling out recent infections. The IgG-positive samples, including weak positives, were further tested in a serum neutralisation test for neutralising antibodies and 54.5% of these samples tested positive. The results show that the virus is in circulation in central and northern regions of Burkina Faso, suggesting the need for improved surveillance and control systems to prevent future outbreaks and the consequent economic impact of the disease in Burkina Faso livestock.

Water requirements for livestock production: a global perspective.

Water is a vital but poorly studied component of livestock production. It is estimated that livestock industries consume 8% of the global water supply, with most of that water being used for intensive, feed-based production. This study takes a broad perspective of livestock production as a component of the human food chain, and considers the efficiency of its water use. Global models are in the early stages of development and do not distinguish between developing and developed countries, or the production systems within them. However, preliminary indications are that, when protein production is adjusted for biological value in the human diet, no plant protein is significantly more efficient at using water than protein produced from eggs, and only soybean is more water efficient than milk and goat and chicken meat. In some regions, especially developing countries, animals are not used solely for food production but also provide draught power, fibre and fertiliser for crops. In addition, animals make use of crop by-products that would otherwise go to waste. The livestock sector is the fastest-growing agricultural sector, which has led to increasing industrialisation and, in some cases, reduced environmental constraints. In emerging economies, increasing involvement in livestock is related to improving rural wealth and increasing consumption of animal protein. Water usage for livestock production should be considered an integral part of agricultural water resource management, taking into account the type of production system (e.g. grain-fed or mixed crop-livestock) and scale (intensive or extensive), the species and breeds of livestock, and the social and cultural aspects of livestock farming in various countries.

Does thyroid-sparing total laryngectomy decrease the risk of hypothyroidism?

BACKGROUND: Thyroid lobectomy is recommended with total laryngectomy for laryngeal cancer in the National Comprehensive Cancer Network ('NCCN') guidelines. However, it is associated with a 32-89 per cent risk of hypothyroidism, with or without adjuvant radiotherapy. OBJECTIVE: The study aimed to determine whether preserving the whole thyroid, compared to a single lobe, does indeed significantly lower the incidence of hypothyroidism in the setting of total laryngectomy. METHOD: A retrospective study was conducted at Groote Schuur Hospital in Cape Town, South Africa. RESULTS: Eighty-four patients met the inclusion criteria. The overall incidence of hypothyroidism was 45.2 per cent. The incidence of hypothyroidism was significantly reduced in patients who underwent thyroid-sparing total laryngectomy compared to hemithyroidectomy (p = 0.037). Adjuvant radiotherapy was associated with a higher incidence of hypothyroidism (p = 0.001). CONCLUSION: Thyroid-preserving laryngectomy should be advocated in carefully selected patients with advanced laryngeal carcinoma, as it reduces the incidence of hypothyroidism.

New developments in the diagnosis of avian influenza.

Avian influenza has become a serious concern from both veterinary and public health points of view. National and international organisations, veterinary health authorities, research institutions, diagnostic laboratories and field services make enormous efforts worldwide to detect, combat and prevent this important disease. Accordingly, the standard diagnostic protocols are being supported by a wide variety of molecular detection techniques, including improved polymerase chain reaction assays, microarray-based detection and characterisation methods, very rapid sequencing, simple pen-side tests and other on-site approaches. These recently developed 'closer to the field' methods allow rapid detection of influenza viruses and the identification of pathogenicity variants. However, in order to harmonise the diagnosis worldwide, attention has to be paid to the validation and standardisation of these technologies, to avoid erroneous interpretation of assay results, and, consequently, inappropriate epidemiological measures. This review gives an overview of the current and potential future developments related to avian influenza diagnostics.

Veterinary vaccines and their use in developing countries.

The burden of infectious diseases in livestock and other animals continues to be a major constraint to sustained agricultural development, food security, and participation of developing and in-transition countries in the economic benefits of international trade in livestock commodities. Targeted measures must be instituted in those countries to reduce the occurrence of infectious diseases. Quality veterinary vaccines used strategically can and should be part of government sanctioned-programmes. Vaccination campaigns must be part of comprehensive disease control programmes, which, in the case of transboundary animal diseases, require a regional approach if they are to be successful. This paper focuses on the salient transboundary animal diseases and examines current vaccine use, promising vaccine research, innovative technologies that can be applied in countries in some important developing regions of the world, and the role of public/private partnerships.

Aspects of kit validation for tests used for the diagnosis and surveillance of livestock diseases: producer and end-user responsibilities.

The Joint Food and Agriculture Organization/International Atomic Energy Agency (IAEA) Division of Nuclear Techniques in Food and Agriculture, based at the IAEA in Vienna, Austria, has extensive experience in helping to develop and validate assays and has provided strong support in developing World Organisation for Animal Health (OIE) norms. This paper will focus on enzyme-linked immunosorbent assay and polymerase chain reaction as the major technologies exploited in diagnosis and surveillance. Problems involving the terminology and factors in kit production, supply and validation are examined, in particular emphasising the importance of robustness and ruggedness of tests. The authors discuss the responsibilities of the various stakeholders (producers, distributors, users, and national/international organisations) in achieving quality controlled data to solve diagnostic and surveillance problems. The roles of internal quality control (internal proficiency testing) and external quality assurance (external proficiency testing) as well as aids to solving problems with kits are examined.

Discrimination between sheep-associated and wildebeest-associated malignant catarrhal fever virus by means of a single-tube duplex nested PCR.

A single-tube duplex nested polymerase chain reaction (sdn-PCR) was developed for the detection of and discrimination between ovine herpesvirus-2 (OvHV-2) and alcelaphine herpesvirus-1 (AIHV-1). These viruses respectively cause sheep- and wildebeest-associated malignant catarrhal fever (SA-MCF and WA-MCF). In the first step of the sdn-PCR, two primers with high annealing temperatures based on conserved regions of the tegument genes were used for DNA amplification. In the second step, two primer sets based on variable regions of the respective OvHV-2 and AIHV-1 genes and with annealing temperatures > 11 degrees C below the primers used in the first step, were used. Internal regions of different sizes from amplicons produced in the first step were amplified. This single-tube test obviates the need for two separate assays to detect both viral types, thereby reducing time, labour and cost.

Eliciting antigen-specific egg-yolk IgY with naked DNA.

Immunization with naked DNA was used to elicit chicken egg yolk antibodies (IgY). Layer hens were inoculated with plasmid DNA encoding the enhanced green fluorescent protein, the fusion protein of Newcastle disease virus, and VP2 of African horse sickness virus. IgY was extracted from egg yolks by polyethylene glycol precipitation. Specific antibodies were present in the yolks of eggs from hens immunized with each of the three different plasmids. This approach to raising polyclonal antibodies obviates the need to produce and purify large quantities of proteins for immunization and can potentially yield large amounts of diagnostically or therapeutically useful reagents.

Direct identification of recombinant vaccinia virus plaques by PCR.

A fast method for the identification of recombinant vaccinia viruses directly from individual plaques is described. Plaques are picked, resuspended in PBS-A and processed for PCR using two 'universal' primers. The amplified sequences are analyzed by agarose gel electrophoresis. This procedure allows discrimination between spontaneously arising TK-negative mutants, which do not carry the inserted gene, and the desired TK-negative recombinants resulting from insertional inactivation of the TK gene.

A comparison of different genomic probes in the detection of virus-specified RNA in Orbivirus-infected cells.

Different 32P-labelled genomic probes of bluetongue virus (BTV), epizootic haemorrhagic disease virus (EHDV) and equine encephalosis virus (EEV) were compared with respect to the detection of virus-specified RNA in infected cells. The probe derived from the genome segment that encodes nonstructural protein NS1 was found to be the most sensitive, detecting virus-specified RNA in glutaraldehyde-fixed cells as early as 2-3 h p.i. This comparison was based on the observation that the NS1 gene probe required a smaller number of infected cells to produce a positive hybridization signal than the other nucleic acid probes. The only exception was the EHDV NS2 gene probe which appeared to be as sensitive as the NS1 gene probe. The advantage of using the NS1 gene probe was particularly evident in the analysis of cells infected at very low multiplicities of infection. At a multiplicity of infection of 1 x 10(-5) plaque forming units/cell, virus-specified RNA could be detected 48 h after infection. The greater sensitivity of the NS1 gene-specific probe is ascribed to the fact that its target, the NS1 mRNA, is transcribed more frequently than the other target viral mRNAs. The major application of the cell-hybridization method is the rapid detection of small quantities of infectious virus particles.

Sequence-independent amplification and cloning of large dsRNA virus genome segments by poly(dA)-oligonucleotide ligation.

A strategy was developed for sequence-independent synthesis and amplification of full-length cDNA of 3-4 kb genes of dsRNA viruses. The method of single primer amplification (Lambden et al., 1992) was adapted by the inclusion of a 3' poly(A) tail to an oligonucleotide ligated to dsRNA genome segments as a template for oligo(dT)-primed cDNA synthesis. Full-length copies of the largest genome segments, 1 (4 kb) and 2 (3 kb), of African horse sickness virus (AHSV) have been cloned, terminally sequenced and expressed in vitro.

The prevalence, risks, and management of Chlamydia trachomatis infections in fertile and infertile patients from the high socioeconomic bracket of the South African population.

OBJECTIVE: The main objective of the study was to evaluate the prevalence of Chlamydia trachomatis endocervicitis in an infertile population. DESIGN: Forty consecutive patients were enrolled in the study group and 41 in the control group. SETTING: The study was undertaken in the Department of Obstetrics and Gynaecology of the University of the Orange Free State, Bloemfontein, Republic of South Africa. PATIENTS: Infertile white females, visiting an infertility clinic in an academic hospital and fertile white female patients visiting an antenatal clinic. INTERVENTIONS: Endocervical swabs were taken, and monoclonal direct immunofluorescence for C. Trachomatis were done on each. MAIN OUTCOME MEASURES: A difference was expected between the prevalence of C. trachomatis infection in the fertile and infertile population. RESULTS: In the study group, 14(35.9%) positive, 25(64.1%) negative, and 1 fallout were obtained. In the control group, 3 patients (7.32%) tested positive. CONCLUSION: Although no correlation was found between C. trachomatis infection of the female genital tract and the clinical history, it showed a significant correlation with infertility. This justifies routine screening tests and antibiotic treatment of positive infertile couples. Analysis of cost-effectiveness showed that empirical treatment of new infertile couples is justified in some populations.

PIP: Health workers at the Department of Obstetrics and Gynecology of the University of the Orange Free State in Bloemfontein, South Africa enrolled 40 consecutive infertile white couples 41 consecutive pregnant white females into a case control study to determine the prevalence of Chlamydia trachomatis infections in an infertile population. Both groups were from the middle to upper socioeconomic class. Laboratory personnel used the monoclonal direct immunofluorescence test to each cervical cytology smear. They had to repeat the test on 5% of the smears. Prevalence of C. trachomatis in the study group stood much higher than it did in the control group (35.9% vs. 7.3%; p.002). No association existed between clinical history and presence of C. trachomatis in the fertile group. 19.5% of the fertile patients had taken antibiotics during the 3 months prior to the study. None reported earlier episodes of salpingitis and/or pelvic inflammatory disease. The researchers proposed a possible reason for the very high rate of C. trachomatis in infertile patients. Perhaps the infertile clinic only examined unresolved infertile cases who may have had an exceptionally high rate of C. trachomatis. The infertility clinic chose to treat all new couples with lymecycline because studies showed that it is always effective against C. trachomatis. Indeed this treatment proved to be the most beneficial at the lowest cost.

Isolation of a neurotoxin from the salivary glands of female Rhipicephalus evertsi evertsi.

A quantitative study of the changes in the protein pattern of the salivary glands of female Rhipicephalus evertsi evertsi during the entire repletion process was undertaken. These results, in conjunction with the previously determined toxic phase, indicated the presence of a toxic protein. The development of a sensitive in vitro assay using a Xenopus nerve-muscle preparation, made it possible to identify toxic phases during feeding and to assay fractions of salivary gland extracts during toxin isolation. Sufficient amounts of electrophoretically and chromatographically homogeneous toxin could be obtained through the use of chromatofocusing, enabling its characterization with respect to molecular weight (68 kDa; determined by gel permeation chromatography), pI (6.00), and amino acid composition. The toxin was inactivated by pronase digestion as well as by antiserum.

Early and late transcriptional phases in the replication of lumpy-skin-disease virus.

Approximately 71 of the estimated 145 kilobase pairs of the genome of the South African isolate of lumpy-skin-disease virus, Neethling strain, was cloned into the plasmid vector pBluescribe. Selected clones were used in Northern blot analysis to investigate the replication cycle of the virus. The synthesis of early mRNA was initiated immediately after infection, and continued for approximately 9 h. The transition to late-gene transcription occurred approximately 10 h post-infection and required DNA replication. Transcription of endogenous late genes cannot occur when viral DNA replication is prevented, while it is activated after DNA replication has occurred. The cloned fragments of the genome are expected to help in the identification of important promoter elements.

Detection of African horsesickness virus and discrimination between two equine orbivirus serogroups by reverse transcription polymerase chain reaction.

A reverse transcription polymerase chain reaction (RT-PCR), based on the gene encoding the NS2 protein of African horsesickness virus (AHSV), was developed for rapid serogroup-specific detection of AHSV. The specificity of RT-PCR products was confirmed by Southern blot hybridization using a radioactively labelled cDNA probe specific for the NS2 gene. This RT-PCR could discriminate between all known members of the AHSV and equine encephalosis virus serogroups. AHSV RNA was detected in a sample representing 0.005 plaque forming units in a dilution series made of infected cell culture material. In an immune horse which had been vaccinated with a baculovirus expressed AHSV (serotype 4) VP2 subunit vaccine, viral RNA could be detected for up to 22 weeks post challenge. AHSV RNA was detected in various organs of an infected horse. Viral RNA was also detected by RT-PCR in nine suspected field cases of African horsesickness while virus isolation was successfully performed on eight of these cases.

Immune responses in a horse inoculated with the VP2 gene of African horsesickness virus.

The ability of a DNA vaccine to elicit an immune response in a horse was evaluated. The outer capsid protein VP2 of African horsesickness virus is known to elicit protective immunity in horses. Reverse transcribed DNA of the gene encoding VP2 was placed under the transcriptional control of the cytomegalovirus immediate-early enhancer/promoter and was injected on several occasions intramuscularly into a horse. Low antibody levels could be detected by ELISA. Antibodies directed against VP2 alone were shown by Western blot while low levels of neutralizing antibodies were detected by a 50% plaque reduction assay. In contrast to a relatively poor humoral response, a significant lymphoproliferative response in the presence of whole virus proteins, as well as a cytotoxic cellular reaction against virus-infected syngeneic target cells was shown.

Characterization of a pigeon paramyxovirus (PPMV-1) isolated from chickens in South Africa.

A paramyxovirus with a thermostability of 60 min (typical of velogenic viruses) and a mean death time of > 90 h (typical of lentogenic viruses) was isolated from layers near Mooi River, South Africa. Our results, based on comparative nucleotide sequence data indicated that the virus is pigeon paramyxovirus 1 (PPMV-1), a variant of Newcastle disease virus. The F0 cleavage site contains a 112RRKKRF117 motif, and the virus had 98% sequence identity with PPMV-1 strains from the Far East. PPMV-1 was last reported in South Africa during the 1980s, with this being the first report of PPMV-1 isolated from chickens in South Africa.

Development of a diagnostic one-tube RT-PCR for the detection of Rift Valley fever virus.

Diagnosis of Rift Valley fever (RVF) is based on serology and virus isolation. The disadvantages of the former include poor sensitivity, high cost, risks associated with using infectious virus as antigen, the lengthy duration of ELISA as well as cross-reactivity with other Phleboviruses. We developed, optimised and evaluated a one-tube reverse-transcription-polymerase chain reaction (RT-PCR) for the detection of Rift Valley fever virus (RVFV) in ruminants. The PCR primers for this assay were designed to anneal to a region within the M segment of the virus genome, encoding glycoproteins G1 and G2. A PCR amplicon of 363 bp was obtained. The sensitivity of the assay was determined to be 0.25 TCID50. This test should allow for the early and rapid detection of RVFV in both serum and whole blood. In addition, it could facilitate the quantification of antigen for the manufacture of current vaccines.

Purification of Cowdria ruminantium by density gradient centrifugation.

The isolation of Cowdria ruminantium by differential and isopycnic density gradient centrifugation is reviewed with special reference to the suitability of Percoll as density gradient medium. Infected sheep brain, Amblyomma hebraeum nymphae and various mouse organs were used as starting material. By these methods, partially purified viable populations of the organism with distinctly different densities were obtained. The conclusions are based upon results of analyses of density fractions by inoculation into sheep or mice, protein determination, electron microscopy and enzyme-linked immunosorbent assay. Morphological differences were observed in the density fractions obtained from infected brain tissue and A. hebraeum.