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Bovine interleukin 2: production kinetics in normal and in vivo antigen-primed cattle.

Conditions influencing production kinetics of bovine interleukin 2 (IL-2), viz. cell concentration, mitogen and its concentration, length of incubation, nutrient medium and in vivo antigen-priming were investigated. Peripheral blood mononuclear cells (PBL) of outbred cattle of different age groups showed considerable variation in their ability to secrete IL-2 which possibly reflects their immune competence. Of the cultures initiated with PBL, 5 x 10(6) cells/ml cultured in serum free Iscove's medium and stimulated with 5 micrograms Con A/ml for 24 hr produced maximal amount of IL-2 activity. In vivo antigen-priming of bovine lymphocytes with the live attenuated rinderpest virus revealed that IL-2 production was not affected by rinderpest virus but the in vivo antigen-priming possibly resulted in concomitant production of suppressor factor(s) which suppressed the already produced IL-2. The implications of this factor(s) in relation to regulation of immune responses in the disease process are discussed.

Animals↗

Serological studies with the virus of peste des petits ruminants in Nigeria.

It was possible to distinguish separate serological responses when experimental goats were inoculated with either rinderpest virus or peste des petits ruminants virus. Examination of field samples established that peste des petits ruminants occur commonly in Nigerian sheep and goats although some villages have escaped recent infection. There was no evidence of infection with rinderpest in these animals.

Animals↗

Risk of introducing exotic disease through importation of animals and animal products.

Between 1870 and 1929, nine separate outbreaks of foot and mouth disease (FMD) occurred in the United States of America (USA); additional outbreaks in North America include one in Mexico (1947) and two in Canada (1870 and 1952). In 1930, the United States Congress enacted a law prohibiting importation of live ruminants or swine or fresh meat from these species into the USA from countries affected with FMD or rinderpest. Although the effect of this prohibition may be debated, the USA has remained free of FMD since its enactment. A hidden benefit of this prohibition was probably the limitation on importing other disease agents from countries of the world where FMD was present. As many regions of the world make progress towards the control and eradication of FMD, North America must take greater cognizance of other disease agents with which it has not been concerned to date, as these existed only in regions of the world affected with FMD and/or rinderpest. One of the methods of dealing with these other diseases is by using risk assessment and risk management methodologies. For risk assessment to work, however, the available management technologies must be examined, and levels of risk assigned to match the available technology. The authors explore risk analysis options for the importation of animals and animal products in a manner which will continue to protect the livestock industry in the USA. They also examine the role of veterinary biologicals as a management tool to mitigate the attendant risks.

Animal Diseases↗

Characterisation of the European seal morbillivirus.

The ELISA test originally developed for the detection of serum antibodies to rinderpest virus has been shown to detect cross-reacting antibodies in sera of diseased common and grey seals. Analysis of sera collected from various seal populations is in progress to establish the correlation between different morbillivirus neutralisation tests, ELISA tests and the disease status of the animals. RNA purified from post-mortem tissues removed from diseased seals has been analysed by hybridisation with cloned cDNAs made to various genes of canine distemper, peste des petits ruminants, rinderpest and measles viruses. This study has confirmed the presence of RNA sequences characteristic of a morbillivirus, but shown that the virus is not identical with any known morbillivirus. Work is in progress to determine the nucleotide sequences of clones carrying inserts homologous to morbillivirus genes isolated from cDNA made on a template of infected seal tissue RNA.

Animals↗

The isolation of peste des petits ruminants virus from northern India.

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.

Animals↗

Cloning of the nucleocapsid protein gene of peste-des-petits-ruminants virus: relationship to other morbilliviruses.

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.

Amino Acid Sequence↗

Detection of peste des petits ruminants virus antigen in conjunctival smears of goats by indirect immunofluorescence.

Peste des petits ruminants virus (PPRV) antigen was detected in conjunctival epithelial cells obtained from goats in the early or late stage of the disease by the use of a specific monoclonal antibody (mAb) to PPRV in an immunofluorescent antibody test (IFAT). The affected goats were sampled during an outbreak of peste des petits ruminants in Eritrea. Syncytia were also observed in some smears, consistent with a morbillivirus infection, but the IFAT was more sensitive than staining for syncytia in the detection of viral antigen, the two tests giving 63 per cent and 40 per cent of animals, respectively, with positive tests. Positive immunofluorescence was observed in samples from goats in the early and late stages of the disease, but was not observed with a specific rinderpest mAb or with conjunctival smears from uninfected animals. It is concluded that preparation of conjunctival smears for staining for syncytia is a simple procedure which can be applied in the field, and by use of a specific mAb PPRV infection can be rapidly confirmed and differentiated from rinderpest.

Animals↗

Classification of peste des petits ruminants virus as the fourth member of the genus Morbillivirus.

Peste des petits ruminants (PPR) is a virus disease of sheep and goats in West Africa. When first described, the virus was considered a variant of rinderpest virus. The biological and physicochemical characteristics of the virus indicate that it is closely related to measles, rinderpest and canine distemper viruses. These three viruses form the genus Morbillivirus of the Paramyxoviridae. PPR virus is sufficiently distinct from these 3 viruses to justify considering it as the fourth member of the Morbillivirus genus.

Animals↗

[Significance of paramyxovirinae protein F in physiopathology and immunity].

The Paramyxovirinae sub-family contains viruses of major importance for humans (measles and mumps viruses) and animals (Newcastle disease virus, canine distemper virus, rinderpest virus) but also zoonotic viruses (Hendra and Nipah viruses). Newcastle disease virus, a Rubulavirus, which is specific of birds, can be used as a model for understanding the pathogenicity mechanisms of Paramyxovirinae and for the study of vaccination against the diseases they cause. The F protein of pathogenic strains of Newcastle disease virus have a polybasic cleavage site (K/R-Q-K/R-R) which is specific of furine, a cellular enzyme present in all cellular types, what allows viral multiplication in all cells of the host. Such cleavage site is also present in the F protein of measles and mumps viruses. Antibodies against the F protein are the most effective in the immune response against Newcastle disease virus. The major role played by the F protein in the immunity against Paramyxovirinae arises also from the research performed on other animal (rinderpest and canine distemper viruses) and human (measles virus) viruses. Recombinant and sub-unit vaccines as well as DNA vaccination based on the expression of viral glycoproteins allow effective vaccination of chickens against Newcastle disease. The use of a similar approach could lead to the development of new vaccines against measles or mumps and if necessary to produce vaccines against zoonotic Paramyxovirinae.

Paramyxovirinae↗

Control of peste des petits ruminants: classical and new generation vaccines.

Peste des Petits Ruminants is a highly contagious disease of domestic and wild small ruminants. It seems to be the major constraint in the development of small ruminant production in areas where it is endemic. Included in the list A of the International Zoosanitary Code, it is also part of the FAO EMPRESS programme. Classically, it is characterised by fever, nasal and ocular discharges, diarrhoea, respiratory distress, mucosal erosive lesions and death in 40-80% of acute cases. All these clinical signs, apart from the respiratory symptoms, are very similar to those of rinderpest. The causal agents of both diseases are closely related and belong to the Morbillivirus genus. In the absence of homologous vaccine, and taking advantage of the close relationship between the two viruses, the attenuated tissue culture rinderpest vaccine has been used for a long time to protect small ruminants against PPR. At the end of 1980s, a PPRV strain was successfully attenuated by serial passages in Vero cells. Demonstrated to be very efficient in the protection of sheep and goats against a virulent challenge, this avirulent PPRV is now widely used in the control of PPR. Its thermostability has been dramatically improved by freeze drying. However, animals inoculated with this vaccine cannot be distinguished serologically from those infected with the wild type viruses. With the reverse genetics technique, it is now possible to engineer PPR marker vaccines which can allow such a differentiation. The genes of the PPRV immune protective proteins, the fusion and the haemagglutinin proteins, have been introduced into the genome of a capripox vaccine strain. The generated viruses have proved to be effective as a dual vaccine to protect against two major diseases of small ruminants: peste des petits ruminants and capripox infections.

Animals↗

Viral and bacterial diseases in livestock in Mongolia.

This review focuses on the status of infectious diseases that are serious for animal health and have adverse economic effects in Mongolia. Data presented here are limited due to the lack of published or other easily available documents. Foot-and-mouth disease continues to cause substantial economic losses as exemplified by the outbreak of infection with serotype O PanAsia lineage virus. In the case of the 2001 outbreak, a 65% reduction in export revenues was recorded. In order to ascertain the free status of Mongolia from rinderpest, sero-epidemiological surveillance has been carried out since 2001. In 2004, Mongolia was certified free from rinderpest by Office International des Epizooties (OIE). A sharp rise in both animal and human brucellosis incidence has become a serious problem. Rabies and anthrax remain endemic with occasional human cases. Other prevailing infectious diseases are contagious pustular dermatitis, contagious agalactia, enterotoxemia and pasteurellosis. The current programs for the control of infectious diseases in livestock in Mongolia lack a definite policy that would enable rapid implementation. A large-scale surveillance of infectious diseases in animals and management of appropriate preventive measures are urgently required in Mongolia.

Animals↗

[Robert Koch's unpublished and unfinished experiments on sheep-pox (author's transl)].

The archives of the Robert Koch Institute include a casket with preparations and handwritten notes by Robert Koch (Fig. 1). He made these preparations during his time as a rural doctor between October of 1878 and September of 1880. They refer to an outbreak of sheep-pox at Rackwitz, a place near his practice at Wollstein (Fig. 2). This work has not been published; we know of it from one of Robert Koch's private letters (11). To reconstruct his working scheme and reasoning, we consulted particularly his reports on rinderpest experiments which he began in 1896 (6). The preparations from this casket which had been stained with Bismarck brown (according to Weigert) date back to a period when Robert Koch developed the foundations of bacteriology and they are evidence of his preparedness to accept new operational procedures (1, 2, 3). Thus, we have to assume that these preparations were to serve as evidence of a bacteriological etiology of sheep-pox. A wrong conclusion as to associations between the superinfection present and etiology of the disease (7) was ruled out by maintaining his own postulate. Simultaneously with this preparation work, Robert Koch performed animal experiments (11). His experience from these studies was utilized later on in his rinderpest experiments (6). On account of his confrontation with viral disease - which had its starting point in his unpublished work on sheep-pox - Robert Koch stated his postulate to be valid in the same manner as if bacteriological etiology had been demonstrated (4,6). The importance of these preparations is also seen in the interpretation of viral tissue damage, i.e. increase of macrophages and plasma cells with subsequent necrosis (9) characteristic of vira infection (Figs. 3, 4).

Animals↗

One-step multiplex RT-PCR assay for the detection of peste des petits ruminants virus in clinical samples.

A single-tube one-step multiplex RT-PCR was standardized to amplify both 337 bp and 191 bp fragments of N and M genes of peste des petits ruminants virus (PPRV), respectively, and only a 337 bp fragment of N gene of Rinderpest virus (RPV). The RT-PCR using purified viral RNA was easily adopted for direct detection of PPRV in clinical field samples and its differentiation from RPV. The amplified N and M gene products were confirmed to be PPRV- and RPV-specific by their size in 1.5% agarose gel and restriction analysis. In the assay, the Qiagen one-step RT-PCR kit containing the Ominiscript and Sensiscript reverse transcriptases and Hot star Taq DNA polymerase was utilized. The sensitivity of the assay was found to be 100 fg of PPRV RNA. Compared with a two-step assay, the one-step assay is easier and time-saving as it requires just a single buffer for both reactions, reverse transcription (RT) and PCR. In experimentally infected goats, PPRV was detectable by the one-step RT-PCR in nasal and ocular swabs 7-17 days post infection (p.i.). and in oral swabs 7-15 days p.i. Out of 32 clinical field samples tested, 18 were positive by sandwich ELISA (S-ELISA), while 22 were positive by the one-step RT-PCR.

Animals↗

The epidemiology of peste des petits ruminants in the Sultanate of Oman.

Virological and serological evidence was obtained to show that peste des petits ruminants virus was widely distributed in Omani sheep and goats. There was no evidence for the concomitant presence of rinderpest virus in these species. Two virus isolates were classified as peste des petits ruminants virus on the basis of their pathogenicity in experimental animals and their specific hybridisation with nucleic acid probes. However, neutralisation tests and polyacrylamide gel analysis of their nucleocapsid proteins showed that they were not identical to the highly conserved African strains of this virus.

Age Factors↗

Identification of T-helper and linear B epitope in the hypervariable region of nucleocapsid protein of PPRV and its use in the development of specific antibodies to detect viral antigen.

Peste des petits ruminants is a highly contagious viral disease of small ruminants making its diagnosis difficult from the similar symptoms of Rinderpest. Computer based prediction algorithms was applied to identify antigenic determinants on the nucleocapsid (N) protein of PPRV. Specificity and antigenicity of each peptide was evaluated by solid phase ELISA. Six specific peptide sequences were evaluated in multiple antigenic peptide (MAP) form and immune response was evaluated by supplementing universal T-helper epitope human IL-1beta peptide (VQGEESNDK, amino acids 163-171). Out of the six peptides 19mer sequence corresponding to 454-472 region of N protein of PPRV was found to be highly immunogenic and specific to PPRV. Evaluation of overlapping peptides differing in length for this 452-472 region, showed minimum length of 14 amino acid residues were required for the stable affinity binding of antigen-antibody. The results of immunization and indirect ELISA indicated the presence of T-helper epitope at the N-terminal end and linear B epitope at the C-terminal region of 454-472 19mer of nucleocapsid peptide of PPRV-nucleocapsid protein. The antipeptide antibodies developed against this region showed specificity to PPRV antigen differentiating it from RPV when used in indirect ELISA and western blot analysis.

Amino Acid Sequence↗

Matrix protein and glycoproteins F and H of Peste-des-petits-ruminants virus function better as a homologous complex.

The matrix (M) protein of paramyxoviruses forms an inner coat to the viral envelope and serves as a bridge between the surface glycoproteins (F and H) and the ribonucleoprotein core. Previously, a marker vaccine (RPV-PPRFH) was produced for the control of peste des petits ruminants (PPR) disease, where the F and H genes of Rinderpest virus (RPV) were replaced with the equivalent genes from Peste-des-petits-ruminants virus (PPRV); however, this virus grew poorly in tissue culture. The poor growth of the RPV-PPRFH chimeric virus was thought to be due to non-homologous interaction of the surface glycoproteins with the internal components of the virus, in particular with the M protein. In contrast, replacement of the M gene of RPV with that from PPRV did not have an effect on the viability or replication efficiency of the recombinant virus. Therefore, in an effort to improve the growth of the RPV-PPRFH virus, a triple chimera (RPV-PPRMFH) was made, where the M, F and H genes of RPV were replaced with those from PPRV. As expected, the growth of the triple chimera was improved; it grew to a titre as high as that of the unmodified PPRV, although comparatively lower than that of the parental RPV virus. Goats infected with the triple chimera showed no adverse reaction and were protected from subsequent challenge with wild-type PPRV. The neutralizing-antibody titre on the day of challenge was approximately 17 times higher than that in the RPV-PPRFH group, indicating RPV-PPRMFH as a promising marker-vaccine candidate.

Animals↗

Optimal time for vaccination against Peste des petits ruminants (PPR) disease in goats in humid tropical zone in southern Nigeria.

Investigation of the incidence of peste des petits ruminants (PPR) disease in goats revealed seasonality in the natural occurrence of PPR disease in the environment. Tissue culture rinderpest vaccine was shown effective in protecting goats against the disease. The optimal time for vaccinating goats against PPR disease was when the least number of animals were incubating the disease. In the tropical humid zone, as in West Africa, the optimal time for vaccination against the PPR disease is from late November to middle of December.

Animals↗

Evidence of identification of peste des petits ruminants from goats in Egypt.

Peste des petits ruminants virus was isolated from young goats in an outbreak of the disease for the first time in Egypt. Affected goats showed symptoms simulating rinderpest, disease in cattle. The mortality rate was about 30%, and morbidity reached 90%. History, clinical symptoms, postmortem lesions, and diagnosis were discussed.

Animals↗