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M W Shirley

Publications and source records attributed to M W Shirley.

At least 37 records · Page 2Linked to original sources

Live attenuated vaccines against avian coccidiosis: Success with precocious and egg-adapted lines of Eimeria.

Avian coccidiosis, caused by parasites of the genus Eimeria, is a major disease of intensively reared poultry. Anticoccidial drugs, and vaccines based on live wild-type (virulent) parasites, have aided the management of coccidiosis, but the development of vaccines based on live attenuated parasites has been a significant advance. Here, Martin Shirley and Petr Bedrník explain the basis of these recent vaccines and discuss their use.

Journal Article↗

Eimeria spp. from the chicken: occurrence, identification and genetics.

Many fundamental aspects of the biology of Eimeria spp. from the chicken remain poorly understood and some have not been investigated in detail for many years. New molecular tools are now available that could be used to underpin some of the more practical aspects of disease control. For example, a far better understanding of the epizootiology of the parasites, with precise knowledge of the characteristics of individual species and strains that are prevalent in the field might, in the future, be incorporated into strategies that see a more rational use of the available drugs and vaccines. The recent use of electrophoretic variation of enzymes to investigate parasite epizootiology is described and the general case for the development of specific, more sensitive DNA-based technologies is discussed. A wealth of DNA probes is potentially available from the large genome of Eimeria spp. and the genetic complexity of these parasites has recently been illustrated in more detail. In addition to a large nuclear genome comprising at least 14 linear chromosomes, studies on E. tenella have shown that, like Plasmodium and Toxoplasma, it also possesses a mitochondrial genome and a newly discovered "plant-like" genome that probably resides within an uncharacterised organelle and was acquired when an ancestor engulfed and kept and algal cell containing a chloroplast. A double-stranded RNA genome has also been identified in some species.

Animals↗

Eimeria tenella: infection with a single sporocyst gives a clonal population.

Two genetically defined parents of Eimeria tenella, each characterized by a selectable phenotype, were mated to provide "recombinant' oocysts for an analysis of the inheritance of DNA markers into the sporocyst stage of the life-cycle. A total of 23 populations was established from infection with a putative single sporocyst and their inheritance of DNA markers provided compelling evidence that the 2 sporozoites within each sporocyst are genetically identical. EAch of the 5 polymorphic DNA markers used segregated independently within the "recombinant' parasites and only 1 of the parental forms was inherited in 22 of the populations aimed to have been established from a single sporocyst.

Adenine↗

Eimeria tenella: genetic recombination of markers for precocious development and arprinocid resistance.

A mating was made between two populations of Eimeria tenella possessing the complementary traits of normal development (virulence) + resistance to the anticoccidial drug arprinocid or precocious development (attenuation) + drug-sensitivity. A small number of "recombinant" oocysts was recovered. The inheritance of markers from both parents into 22 cloned lines derived from the recombinant oocysts was confirmed by analyses of restriction fragment length polymorphisms of four repetitive DNA sequences.

Adenine↗

A live attenuated vaccine for the control of avian coccidiosis: trials in broiler breeders and replacement layer flocks in the United Kingdom.

Losses caused by coccidiosis are a significant problem in the rearing of breeder and layer flocks. A live vaccine has been developed that contains attenuated lines of the seven species of Eimeria that infect the chicken. The attenuated lines were derived from virulent strains by selection for earlier development in chickens. In 11 field trials, the performance of vaccinated chicks was compared with that of matched controls receiving conventional drug prophylaxis. The vaccine was given in the drinking water to 116,600 young chickens and provided excellent control of coccidiosis. The occurrence of coccidial oocysts in the litter, coccidial lesions post mortem and overt coccidiosis was markedly lower in the vaccinated birds than in the controls.

Animals↗

The genome of Eimeria tenella: further studies on its molecular organisation.

The molecular karyotype of the coccidial parasite Eimeria tenella has been further resolved. Different conditions of pulsed-field gel electrophoresis were used to separate up to 14 chromosomes that ranged in size between 1 and in excess of 6 megabase pairs (Mb) of DNA. Comparisons of several populations revealed some size polymorphisms of homologous chromosomes. Probes containing tandem repeats of the DNA sequence CAG and the telomeric-like sequence TTTAGGG hybridised to each chromosome. All other probes containing either repetitive or non-repetitive sequences were mapped to individual chromosomes.

Animals↗

Intra-specific variation within Eimeria tenella detected by the random amplification of polymorphic DNA.

The random amplification of polymorphic DNA (RAPD method) was used to derive fragments of DNA of the coccidial parasite Eimeria tenella for use both as genetic markers and as an aid for the discrimination of different wild-type and attenuated populations. Of 41 arbitrarily chosen 9- to 27-mer primers, 24 yielded useful arrays of fragments following low-stringency annealing conditions and the resultant profiles were generally very reproducible. One non-variant fragment of 2 kb hybridised to a single chromosome (number 12) and four variant fragments were identified. These results strongly suggest that the RAPD method may be an extremely useful tool for studies on various aspects of the genetic organisation of coccidial parasites.

Animals↗

Coccidial parasites from the chicken: discrimination of different populations of Eimeria tenella by DNA hybridisation.

Four DNA probes that are reiterated several times within the genome of Eimeria tenella have been used to derive restriction fragment length polymorphism patterns that are characteristic for different laboratory-maintained or recently isolated populations. Three of the four probes hybridised only to DNA from E tenella whereas probe B209 also hybridised to DNA from both E praecox (strongly) and E acervulina (more weakly). The results of this study confirm the usefulness of DNA probes for taxonomic or identification studies and indicate that, with further effort, a wider range of strain-, species- and genera-specific DNA probes could be isolated to expand our understanding of the epizootiology of coccidial parasites.

Animals↗

7H8/6, a multicopy DNA probe for distinguishing isolates of Plasmodium falciparum.

A method using DNA fingerprinting to distinguish Plasmodium falciparum isolates has been developed using a clone, 7H8/6, isolated by screening a genomic expression library of the Malayan Camp strain with a monoclonal antibody. Unlike other P. falciparum DNA probes that contain internally repetitive sequences, this probe contains a single full-length open reading frame and lacks any repetitive sequences. Chromosome mapping located the hybridizing sequences to most chromosomes. 7H8/6 sequences are located at sites near the subtelomeric regions, although they are further from the telomeres than the subtelomeric repetitive sequences rep20. The probe gave multiple polymorphic bands on Southern blots of genomic DNA allowing all unrelated isolates tested to be readily distinguished. Hybridization patterns of parent and progeny from the genetic cross of two P. falciparum clones showed that most bands were inherited independently and no new bands were generated during the cross. This probe is useful for distinguishing isolates of P. falciparum and also for analyzing relationships between closely related isolates.

Animals↗

A molecular karyotype of Eimeria tenella as revealed by contour-clamped homogeneous electric field gel electrophoresis.

DNA from sporozoites of Eimeria tenella was resolved by pulsed field gel electrophoresis into nine chromosomal bands. Some bands of this molecular karyotype contained more than one chromosome as determined by the relative intensity of both staining with ethidium bromide and hybridisation to an E. tenella telomeric probe. Haploid forms of the parasite must be presumed to contain at least 12 chromosomes. The two smallest chromosomes were about 1.1 and 1.4 megabases. Most chromosomes were in excess of 3 Mb with the largest over 5 Mb as determined by comparison with the co-migration of chromosomes from Schizosaccharomyces pombe. A 5S ribosomal gene probe hybridised to a single chromosomal band.

Animals↗

Detection of DNA from infectious laryngotracheitis virus by colourimetric analyses of polymerase chain reactions.

A combination of the polymerase chain reaction and a novel ELISA-type DNA colourimetric assay (developed from studies with a retrovirus from man) was used in a preliminary study to detect DNA from avian infectious laryngotracheitis virus. The method is sensitive, specific and easy to perform. Since it can be readily adapted for the detection of DNA from other sources it could be useful for the identification of a variety of pathogens from other species of veterinary importance.

Animals↗

Chromosome 9 from independent clones and isolates of Plasmodium falciparum undergoes subtelomeric deletions with similar breakpoints in vitro.

We show that chromosome 9 in all isolates and clones of Plasmodium falciparum examined so far exists as one of two distinctly different forms, a large form about 1.9 megabases long or a smaller form about 25% shorter. Physical maps of chromosome 9 from independent clones with large and small forms of chromosome 9, and from an isolate with the large form and 3 derived clones with the small form reveal the underlying structural basis of this size polymorphism. The small form differs from the large only in that there are subtelomeric deletions at each end, one of these deletions involving about 0.45 megabases. Remarkably, the breakpoints map within about +/- 1% of the total chromosome length for each of these populations. We discuss some possible mechanisms for this.

Animals↗

Characterization of coccidial proteins by two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis.

Two dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (2D SDS-PAGE) has been used to produce 'fingerprint' maps of the proteins from each of the 7 species of Eimeria which infect the chicken. All 7 species could be identified from their array of polypeptides but few differences were detected between strains of the same species. Alterations to the polypeptide array associated with the stage of sporulation of the oocysts were observed. Iodination of sporozoites, 2D SDS-PAGE, autoradiography and immunoblotting techniques were combined to identify polypeptides with a surface moiety and those which were antigenic.

Animals↗

Sensitivity of field isolates of Eimeria species to monensin and lasalocid in the chicken.

Isolates of Eimeria species obtained from broiler or from breeder farms were compared for their sensitivity to two ionophorous anticoccidial drugs, monensin and lasalocid. All of 25 isolates from broiler farms were resistant to 100 ppm monensin or 90 ppm lasalocid, while 14 of 16 isolates were resistant to monensin and seven of 16 to lasalocid from breeder farms (replacement layer and broiler breeder).

Animals↗

Enzyme variation and pathogenicity of recent field isolates of Eimeria tenella.

Seventy isolates of Eimeria tenella, obtained from commercial poultry farms worldwide and four reference laboratory strains were characterised by studies on the electrophoretic mobility of up to three enzymes. All populations possessed the same electrophoretic form of lactate dehydrogenase and malate dehydrogenase and one of two forms of glucose phosphate isomerase. One isolate was characterised by both forms of glucose phosphate isomerase. Studies on several isolates indicated that there was no correlation between the form of glucose phosphate isomerase found and the pathogenicity of an isolate.

Animals↗