Biomedical subjects
Publications and source records attributed to A Diallo.
The authors report 20 cases of trauma of the male external genital organs observed over a 10-year period, due to various causes, particularly road accidents, domestic quarrels, and self-mutilation. Lesions of the testis and its appendages were frequent and treatment was usually surgical.
Peste des petits ruminants (PPR) is an important viral disease of goats and sheep prevalent in West Africa and the Middle East. In recent years, PPR has emerged in India, first in the South India and later in North India. To study the genetic relationships between viruses of distinct geographical origin we have sequenced a 322 nucleotide cDNA fragment of the fusion protein gene generated using reverse transcription followed by polymerase chain reaction (PCR) amplification. Viruses from nineteen independent PPR outbreaks were compared; these included the prototype African strain from Senegal and viruses from disease outbreaks which have occurred at different times and locations across Africa, Arabia, the Near East and the Indian subcontinent. Four separate lineages of the virus were identified and the virus isolates from Asia over the past 2 years were all of one lineage which had not previously been identified in Africa or Asia.
The aetiological agent responsible for an epizootic of a rinderpest-like disease afflicting sheep and goats in three states of northern India was confirmed as peste des petits ruminants virus. To differentiate the virus from rinderpest a number of diagnostic tests were used, including immunocapture ELISA, specific oligonucleotide primers in a reverse transcriptase polymerase chain reaction, immunofluorescence with virus specific monoclonal antibodies and virus isolation. The virulence profile of one isolate in cattle sheep and goats was established. Infected animals developed specific antibody responses and excreted specific antigen in their lachrymal secretions.
Two cases of vitrectomized congenital chorioretinal toxoplasmosis are presented. During a recurrence, an anterior segment inflammation with hypertony complicated both cases. Hypertony was efficiently controled by a specific antitoxoplasmosis therapy. The role of the vitreous barrier is suggested.
Within the epidemiological surveillance of rinderpest in Mali a serological survey has been carried out on 58 herds of small ruminants. Out of 567 tested sera for the detection of antibodies against rinderpest 2 were positive. These sera were collected from two animals over 6 years old, probably infected during the last outbreak of rinderpest in Mali in 1986. Therefore, it can be assumed that the rinderpest virus has not circulated in Mali since that year. However, the infection rate among goats and sheep due to the PPR virus seemed to be high: 74% of herds had already been infected. The prevalence of individual infection is 32%. A similar serological survey was conducted on 450 cattleheads, without antibodies against the rinderpest virus and showed that 1.78% of these animals had been in contact with the PPR virus. With such a low infection rate in cattle, the PPR virus probably has no incidence in the epidemiology of rinderpest in Mali.
The ability of the attenuated peste des petits ruminants vaccine virus to protect small ruminants against virulent rinderpest virus was investigated. Out of four susceptible goats that were infected with the highly virulent Saudi strain of rinderpest virus by intranasal ioculation, three developed mild clinical signs of disease and infected susceptible in-contact goats and cattle with rinderpest virus. However, four goats which had been vaccinated with the attenuated peste des petits ruminants virus resisted challenge with virulent rinderpest virus and did not infect susceptible in-contact animals.
A competitive ELISA based on the reaction between a monoclonal antibody (mAb) and a recombinant nucleoprotein of the peste des petits ruminants virus (PPRV) was developed. This protein was obtained in large quantities from insect cells infected with a PPR nucleoprotein recombinant baculovirus (N-B). The competitive ELISA was compared with the virus neutralisation test (VNT) for detecting specific antibodies to PPRV in sheep and goats. The time consuming VNT is the only prescribed test that is capable of distinguishing between PPRV and the cross-reactive rinderpest virus (RPV). The competitive ELISA involves the simultaneous addition of the mAb and antibodies present in a positive serum, leading to competition for a specific epitope on the N-B. Optimum conditions were obtained by using serum samples which had positive or negative neutralising activity against PPRV or RPV. A negative cut-off point was determined on PPRV-negative sera from RPV-vaccinated cattle. A threshold value of 48 per cent inhibition, calculated from the mean for this population plus 2.7 standard deviations, was used in routine testing. A total of 683 sera were analysed by the competitive ELISA and the VNT. A good correlation (r = 0.94) was observed between the titres obtained in the two tests, with 80 sera that were from laboratory sources. The agreement between the two tests was determined on 271 field sera (kappa = 0.825). Their relative sensitivity (94.5 per cent) and specificity (99.4 per cent) were assessed on the 148 laboratory sera plus the 271 sera used for the determination of kappa.(ABSTRACT TRUNCATED AT 250 WORDS)
cDNA corresponding to the fusion protein (F) gene of the vaccine strain of peste des petits ruminants virus (PPRV) was cloned and sequenced. The gene was 2321 nucleotides long excluding the poly(A) tail. As with other morbilliviruses, it had a long G/C rich stretch of about 525 nucleotides. There was no start codon before position 489 in the nucleotide sequence. From the 489th nucleotide to the 549th nucleotide, there were 4 ATG codons, two of which were in frame. The fourth ATG codon was in the best context to act as a start codon for encoding a protein which will be composed of 546 amino acids with a predicted molecular weight 59,310 Da. The comparison of the nucleic acid sequences of different morbillivirus F-protein genes revealed that the 5'-end sequence of the mRNA is specific to each virus. This sequence contains a long stretch of nucleotides rich in G/C content. When protein sequences were compared, it appears that, during evolution, substitutions in amino acid occurred in the F-protein of morbilliviruses such that the structure required for the fusion activity remains unchanged. These substitutions appear to have only occurred in the leader and the membrane anchor sequences, probably as an adaptation of the protein to the host cell.
Effective implementation of control measures for rinderpest and peste des petits ruminants requires that a proper and rapid diagnosis of the disease is made. Peste de petits ruminants (PPR) can be confused clinically with other infections such as pasteurellosis or contagious ecthyma. Rinderpest, in its classical form, is easy to identify clinically; however, mass vaccination in many countries and also the emergence of mild strains of the virus have made clinical diagnosis more difficult. Clinical observations for both diseases should always be confirmed by a laboratory. Diagnostic techniques used in the past were virus neutralization, agar gel immunodiffusion and virus isolation in cell culture, followed sometimes by reproducing the disease in susceptible animals. All these techniques are either time-consuming, labour intensive, insensitive, or expensive to perform. With the advent of hybridoma and molecular biological techniques, new reagents to assist diagnosis have become available and have led to the development of specific and rapid tests for the diagnosis of each disease. The present article reviews the diagnostic techniques currently available. An indirect ELISA was used successfully to evaluate the status of cattle following the Pan African Rinderpest Campaign. More recently competitive or blocking ELISAs have been developed based on monoclonal antibodies specific for the N or H proteins of the viruses, and which enable differential diagnosis between rinderpest and PPR. This is particularly important in sheep and goats, which may be infected with either virus. In future, improved standardization and reduced costs may be expected with the introduction of ELISAs based on purified antigens expressed in gene vector systems such as baculovirus. ELISA may also be adapted to antigen detection. Nucleic acid technology has also been applied to virus detection procedures. Hybridization probes showed a disappointing sensitivity for diagnostic applications, but more recently the polymerase chain reaction method has shown great promise, providing the potential of high sensitivity combined with specificity.
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An immunocapture ELISA for the diagnosis of rinderpest and peste des petits ruminants is described. Monoclonal antibodies directed against non-overlapping antigenic domains on the nucleocapsid (N) were used to detect the virus N protein in supernatants from infected cells and in field specimens. The assay, which is very sensitive, can be performed in one hour on pre-coated plates. There was no cross reaction between the two viruses in the test and the N protein could be detected in infected cell supernatants kept at ambient temperature for one week. These results show that the ELISA is suitable for routine diagnosis of field samples.
Two independent cDNA clones, identified as representing the mRNA of the nucleocapsid protein gene of peste-des-petits-ruminants virus, were sequenced. The longest insert was 1662 nucleotides, not counting the poly(A) tail, and it was estimated that about 21 nucleotides were missing from the complete gene sequence. The sequence contained one long open reading frame encoding a protein of 525 amino acids with a predicted relative molecular mass of 58,008. Comparisons of the nucleic acid and protein sequences of all the morbillivirus nucleoproteins so far determined indicated two major subgroups in the morbillivirus genus of the Paramyxoviridae: one group included canine and phocine distemper viruses, and the other rinderpest, measles and peste-des-petits-ruminants viruses. Peste-des-petits-ruminants virus was found to be slightly more related to canine and phocine distemper viruses than were measles and rinderpest viruses.
The aim of this study is: 1. to isolate intrahepatic biliary epithelial cells and, 2. to study the toxicity of cholestatic biliary acids on these cells. Cells were isolated from rats with proliferated bile duct-cells, that were induced by a 21 days bile duct ligation. They were characterized by glutamyltranspeptidase and cytokeratins 7 and 19 using histochemical or immuno cytochemical methods. Isolated cells were incubated with different concentrations (250, 500, 1000 and 2000 microM) of cholestatic biliary acids, lithocholate sulfate (LCS) and taurolithocholate sulfate (TLCS. Cells viability is estimated by three methods: Trypan blue, LDH release and MTT test. We obtained purified fractions of non parenchymal liver cells enriched in biliary epithelial cells (> 95%). On these cells, we showed toxicity of LCS and TLCS and determined CI 10 and CI 50 of these acids which were respectively 800 microM and 2 mM for LCS; 1.4 and 2 mM for TLCS. These results indicate that cholestatic biliary acids (LCS and TLCS) are toxic for biliary cells. This cytotoxicity can be probably a possible mechanism of cholestasis.
With a sample of 40 patients, the authors conducted a prospective case-control study on the feasibility of differed autologous blood transfusion at Point "G" National Hospital in Bamako, Mali. The method has been accepted by all the patients. No clinical nor biological trouble has been reported by the patients themselves. The authors recommend the use of autologous blood transfusion by other hospital units and the development of regulations governing its implementation.
In this study we measured biliary cytosolic calcium and examined the effect of cholestatic bile acids LCS and TLCS on intrahepatic isolated biliary cells cytosolic calcium. Cells have been isolated from bile duct ligated rats. Cytosolic calcium has been measured by using the Ca++ sensitive indicator Fura 2 and a cytofluorimetric method. LCS and TLCS (200 microM and 300 microM) increased the cytosolic Ca++ concentration of the cells. In contrast, the bile acids cholate and urso-desoxycholate which are choleretic had no effect. The number of cells which have increased their cytosolic calcium was directly correlated with the biliary acid toxicity. The increase induced by LCS and TLCS was abolished by removing external calcium. It is suggested that the calcium increase results from external calcium influx. This cytosolic calcium increase is known to be toxic for cells so it is concluded that this calcium increase is probably involved in the toxicity of LCS and TLCS.
An exact assessment of the animal health situation in a country is an essential element in formulating eradication and control programmes, and in regulating international trade in animals and animal products from that country. Due to a lack of human and technical resources, Veterinary Services in developing countries often lack precise knowledge on disease occurrence. Since the collection and transmission of reliable information on animal diseases in developing countries are major concerns of the Office International des Epizooties (OIE), a project aimed at improving this situation was implemented with international financial support. This project involved the development by the Centre for the Application of Methodology for the Diagnosis of Animal Diseases (CAMDA) of field kits for the diagnosis of the main diseases present in tropical Africa: rinderpest, peste des petits ruminants (PPR), contagious bovine pleuropneumonia (CBPP) and contagious caprine pleuropneumonia (CCPP). Several tests already exist, such as complement deoxyribonucleic acid (cDNA)-specific probes and polymerase chain reaction (PCR) for rinderpest and PPR, DNA probes and PCR for CBPP, capture enzyme-linked immunosorbent assay, the agglutination test and the immunobinding peroxidase test for CCPP, etc. With specific reference to these examples, the various problems faced by the OIE and CAMDA are reviewed.
A competitive ELISA (C-ELISA) using monoclonal antibodies (mAbs) which bind to the nucleo-protein (NP) of rinderpest virus (RPV) for detection of RPV antibodies in cattle and small ruminant sera is described. Unlike virus neutralisation test (VNT), this test using mAb IVB2-4, can detect specific RPV antibodies without showing a cross-reaction with antibodies to peste-des-petits ruminants-virus (PPRV); by contrast, when mAb VE4-1 is used the test detects both RPV and PPRV antibodies, including low levels of antibodies that can be found in sera containing maternal antibodies. Although antibodies to the PPRV 75-1 strain are also detected with mAb 51-5-6, the test is suitable for assessing the immune status of cattle against the Rinderpest Old Kabete (RBOK) strain. The results from a panel of sera with a known status of vaccination provide evidence for a highly significant correlation between C-ELISA and VNT. This test may be a useful tool for a standardized and accurate determination of the immunity status of both cattle and small ruminants.
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