Biomedical subjects

R C A Thompson

Publications and source records attributed to R C A Thompson.

At least 19 recordsLinked to original sources

PubMed2006

Direct characterization of Blastocystis from faeces by PCR and evidence of zoonotic potential.

In vitro propagation followed by PCR, and a PCR-based method capable of the direct detection of Blastocystis in faeces were utilized to detect Blastocystis from various hosts in Australia, including primates and their handlers from the Perth Zoo. In addition, Blastocystis isolates from dogs and humans living in a localized endemic community in Thailand were also characterized genetically. PCR-based detection directly from faeces was shown to be more sensitive compared with in vitro culture for the detection of Blastocystis. Moreover, phylogenetic analysis of Blastocystis isolates amplified utilizing in vitro techniques prior to PCR revealed that this method favoured the preferential amplification of Blastocystis subtype 5 over subtype 1. This study is the first to provide molecular-based evidence supporting the zoonotic potential of Blastocystis in dogs, possums and primates in a natural setting.

Animals↗

PubMed2006

A case of mistaken identity--reappraisal of the species of canid and felid hookworms (Ancylostoma) present in Australia and India.

This study serves to clarify the current status of canid and felid Ancylostoma species present in Australia. The morphological identification of A. ceylanicum from cats for the first time in Townsville, Australia, appears to be in error, together with the genetic markers provided for the species. Morphological and genetic data presented herein provide strong evidence that the hookworms from cats in Towsville are not A. ceylanicum as previously identified (i.e. the first report of this species in Australia), but are A. braziliense. Therefore the subsequent genetic markers established for A. ceylanicum in subsequent molecular studies based on these Townsville specimens should also be attributed to A. braziliense. Based on this information, a study of canine hookworm species present in northern India is also in error and it is apparent that the hookworms found in this region are those of A. ceylanicum. The distribution of A. braziliense and A. ceylanicum in the Americas and Asia Pacific region is discussed together with the importance of combining parasite morphology with genetic data for parasite diagnosis in epidemiological studies.

Ancylostoma↗

PubMed2006

Comparative development of Echinococcus multilocularis in its definitive hosts.

The comparative development of Echinococcus multilocularis was studied in its definitive hosts, the fox, dog, cat and raccoon dog, beyond the pre-patent period to 90 days post-infection. All host species, apart from cats were susceptible to infection and capable of supporting substantial worm burdens. Although worms in cats matured and produced thick-shelled eggs, their overall development was retarded compared to that in other species in which the parasite matured rapidly producing large populations of gravid worms. E. multilocularis matured rapidly in foxes and raccoon dogs and this was sustained in raccoon dogs but not in foxes in which maturation of worms declined during the later stages of infection, in contrast to that in both raccoon dogs and dogs. These populations were sustained for longer in raccoon dogs and dogs compared to foxes. Cats would appear to have only a minor role in the maintenance of E. multilocularis in endemic areas, and infections in cats may be of minimal public health significance. In contrast, foxes, dogs and the recently recognized definitive host the raccoon dog, are all capable of playing significant roles in the epidemiology of alveolar echinococcosis. This study also demonstrated that the developmental processes of growth, segmentation, proglottization and maturation in adult Echinococcus are independent and can be influenced by environmental factors thus confirming earlier in vitro observations.

Animals↗

PubMed2005

Molecular and morphological characterization of Echinococcus in cervids from North America.

Many issues concerning the taxonomy of Echinococcus have been resolved in recent years with the application of molecular tools. However, the status of Echinococcus maintained in transmission cycles involving cervid intermediate hosts remains to be determined. The recent characterization of the parasite from cervids in Finland has highlighted the paucity of data available, particularly that from North America. In this study, we have characterized a large number of Echinococcus isolates from cervids from Western Canada on the basis of morphology and molecular genetic techniques. Our results support earlier studies suggesting that Echinococcus of cervid origin is phenotypically and genetically distinct to Echinococcus maintained in domestic host assemblages, and also confirms that Echinococcus of cervid origin does not constitute a genetically homogeneous group. However, our data do not support the existence of 2 distinct genotypes (strains/subspecies) with separate geographical distributions. Our data appear to support the existence of only 1 species in cervids, but additional isolates from cervids and wolves in other endemic regions should be characterized before a final decision is made on the taxonomic status of Echinococcus in cervids.

Adenosine Triphosphate↗

PubMed2005

Reproductive potential of Echinococcus multilocularis in experimentally infected foxes, dogs, raccoon dogs and cats.

A total of 15 red foxes, 15 raccoon dogs, 15 domestic dogs and 15 domestic cats were each infected with 20,000 protoscolices of Echinococcus multilocularis. At 35, 63, and 90 days post inoculation (dpi), five animals from each group were necropsied and the worm burdens determined. The highest worm burdens in foxes (mean of 16,792) and raccoon dogs (mean of 7930) were found at 35 dpi. These declined to a mean of just 331 worms in foxes and 3213 worms in raccoon dogs by day 63 with a further decline to 134 worms in foxes and 67 worms in raccoon dogs by day 90. In dogs, there was no significant difference between worm burdens recovered at days 35 (mean of 2466) and day 90 (mean of 1563), although reduced numbers were recovered on day 63 (mean of 899). In cats, worms were found in four animals 35 dpi (mean of 642), in three at 63 dpi (mean of 28) and in two at 90 dpi (mean of 57). Faecal egg counts were determined at 3 day intervals from 25 dpi. A mathematical model of egg excretion dynamics suggested that the mean biotic potential per infected animal was high in foxes (346,473 eggs); raccoon dogs (335,361 eggs) and dogs (279,910 eggs) but very low for cats (573 eggs). It also indicated that approximately 114, 42 and 27 eggs per worm were excreted in the faeces of dogs, raccoon dogs and foxes, respectively. The fecundity of worms in cats was low with an average of less than one egg per worm. The peak levels of coproantigen were detected earlier in foxes and raccoon dogs than in dogs. Eggs recovered from foxes, raccoon dogs and dogs resulted in massive infections in experimental mice. However, metacestodes did not develop from eggs originating from infected cats. It is concluded that foxes, raccoon dogs and dogs are good hosts of E. multilocularis. In contrast, the low worm establishment, the very few excreted eggs and the lack of infectivity of eggs strongly indicate that cats play an insignificant role in parasite transmission.

Animal Diseases↗

PubMed2005

Characterization of Trypanosoma evansi type B.

A distinctive feature of Trypanosoma evansi is the possession of a kinetoplast that contains homogeneous DNA minicircles, but lacks DNA maxicircles. Two major sequence variants of the minicircle have been described and here we have sequenced the type B variant and designed a specific PCR test to distinguish it from type A. Further a test based on maxicircles to distinguish T. brucei brucei from T. evansi was designed and evaluated. Using the designed PCR tests, we detected three type B isolates from camel blood samples collected in northern Kenya, more than 20 years after the first isolation of type B. Comparison of minicircle sequences from all four type B isolates shows >96% identity within the group, and 50-60% identity to type A minicircles. Phylogenetic analysis based on minicircle sequences reveals two clusters, one comprising isolates of type A and one of type B, while random amplification of polymorphic DNA show slight polymorphic bands within type B. Most T. evansi isolates analysed were heterozygous at a repetitive coding locus (MORF2). All type B isolates had one genotype designated 3/5 based on the alleles present. Three camel isolates, which had homogenous type A minicircles, lacked the RoTat 1.2 gene, while another five isolates were T. b. brucei, based on the heterogeneity of their minicircles and presence of maxicircles as demonstrated by PCR amplification of the gene for cytochrome oxidase subunit 1. Our results confirm the existence of T. evansi type B isolates, T. b. brucei and existence of T. evansi type A without RoTat 1.2 gene in Kenyan isolates.

Animals↗

PubMed2005

Emergence/re-emergence of Echinococcus spp.--a global update.

This review provides an update of the biological aspects of the genus Echinococcus and focuses on newly recognized endemic areas. Infection with the intermediate cystic stage of all species of Echinococcus causes disease and incapacity in animals and humans, and in the most serious cases, death of the host. Transmission of Echinococcus to new continents has occurred during European colonisation and the parasite has often taken advantage of Echinococcus-naive wildlife populations in these new environments, incorporating them into its transmission pattern. Echinococcus granulosus consists of a complex of 10 strains. Host specificities of these strains have important implications for transmission and control. As a result of human behaviour and/or political instability in a number of countries Echinococcus is re-emerging as an important public health issue. The importance of wildlife reservoirs in perpetuating transmission and as a source of infection for domestic animals and humans is addressed. The review also refers to the transmission pattern of a recently described new species, Echinococcus shiquicus, from China.

Animals↗

PubMed2005

Cryptosporidium and cryptosporidiosis.

Cryptosporidium is one of the most common enteric protozoan parasites of vertebrates with a wide host range that includes humans and domestic animals. It is a significant cause of diarrhoeal disease and an ubiquitous contaminant of water which serves as an excellent vehicle for transmission. A better understanding of the development and life cycle of Cryptosporidium, and new insights into its phylogenetic relationships, have illustrated the need to re-evaluate many aspects of the biology of Cryptosporidium. This has been reinforced by information obtained from the recent successful Cryptosporidium genome sequencing project, which has emphasised the uniqueness of this organism in terms of its parasite life style and evolutionary biology. This chapter provides an up to date review of the biology, biochemistry and host parasite relationships of Cryptosporidium.

Animals↗

PubMed2004

The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes.

There are 11 different pathogenic trypanosomes in trypanosomiasis endemic regions of Africa. Their detection and characterisation by molecular methods relies on species-specific primers; consequently several PCR tests have to be made on each sample. Primers ITS1 CF and ITS1 BR, previously designed to amplify the internal transcribed spacer (ITS1) of rDNA, have been evaluated for use in a universal diagnostic test for all pathogenic trypanosomes. Blood was collected from 373 cattle and 185 camels. The primers gave constant PCR products with the stocks of each taxon tested. Members of subgenus Trypanozoon (T. brucei brucei, T. evansi, T. b. rhodesiense and T. b. gambiense) gave a constant product of approximately 480 bp; T. congolense, savannah 700 bp, T. congolense kilifi 620 bp and T. congolense forest 710 bp: T. simiae 400 bp, T. simiae tsavo 370 bp, T. godfreyi 300 bp and T. vivax 250 bp. The sensitivity of the test ranged from 10 pg for Trypanozoon, T. congolense clade and T. vivax to 100 pg for T. simiae and T. godfreyi. The primers detected cases of multi-taxa samples, although the sensitivity was reduced with an increase in the combinations. A better detection rate of trypanosome DNA was recorded with buffy coats than from direct blood. With the field samples, the diagnostic sensitivity was close to the sensitivity obtained using single reactions with species-specific primers for Trypanozoon 38/40 (95%) and T. congolense savannah 30/33 (90.9%) but was lower with T. vivax 25/31 (77.4%). The primers offer promise as a routine diagnostic tool through the use of a single PCR; however, further evaluation is recommended.

Animals↗

PubMed2004

Detection of Trypanosoma evansi in camels using PCR and CATT/T. evansi tests in Kenya.

Camel trypanosomosis (Surra) causes high morbidity and is an impediment to the camel husbandry in Kenya. The lack of a sensitive diagnostic test has hindered the collection of accurate epidemiological data and institution of control programmes. A cross-sectional study was conducted in three districts of Kenya to estimate the prevalence of Trypanosoma evansi (T. evansi) and to compare four diagnostic tests: polymerase chain reaction (PCR), card agglutination test (CATT/T. evansi), microhaematocrit centrifugation technique (MHCT) and mouse inoculation (MI). A total of 549 camels were randomly sampled. The overall prevalence of Surra was 5.3% using MHCT, 26.6% using PCR and 45.9% using CATT/T.evansi. There was a significant difference (P < 0.001) between PCR and CATT/T.evansi test, MHCT and MI in detection of T. evansi. The prevalence of T. evansi was 39.8% in Samburu, 24.7% in Nanyuki and 14.4% in Isiolo districts using PCR. A male camel was 2.6 times more likely to be infected with T. evansi compared to a female camel (OR = 3.0% CI: 1.6, 4.1), while an adult camel was 2.2 times more likely to be infected compared to non-adults (OR = 2.2; 95% CI: 1.2, 5.0). There was a poor association between the presence of the published clinical signs and seropositivity (kappa = 0.12), PCR (kappa = 0.11) and MHCT (kappa = 0.05). However, there was a higher agreement between farmers' classification of disease with the PCR test (kappa = 0.5, n = 61). The mean PCV varied with age, presence of infection, locality and gender, with the lowest mean PCV being recorded in MHCT-positive animals (20.97 +/- 0.5) and from infected calves (19.5 +/- 1.2). This study shows that PCR was more sensitive in detecting T. evansi than other tests used. Further, the prevalence of T. evansi in the camel herds sampled is higher than that previously reported in Kenya, and that the judgment by camel keepers may be a reliable "pen-side" diagnostic test for Surra. Considering the low sensitivity of parasitological techniques in detection of chronic T. evansi infection and high cost of PCR, development of a sensitive pen side diagnostic test, with a low cost is still a priority.

Age Factors↗

PubMed2004

Molecular epidemiology of Giardia and Cryptosporidium infections in dairy calves originating from three sources in Western Australia.

A longitudinal study was undertaken to determine the spread of naturally acquired Giardia and Cryptosporidium in dairy cattle in Western Australia. Samples were collected weekly from birth to weaning, and only calves that were sampled four or more times were included in the analysis. It was found that calves rapidly acquire infections with Cryptosporidium parvum and Giardia, with Cryptosporidium being the first to appear within 1-3 weeks after birth whereas Giardia was most prevalent in calves 4-7 weeks of age. Cryptosporidium and Giardia were detected in 48 and 89% of sampled calves, respectively. No significant association was observed in the present study between parasite occurrence and season or management practices (housing), and calf-to-calf contact appears to be the most likely source of transmission. Molecular characterization of isolates of Giardia and C. parvum recovered from calves demonstrated that only the occurrence Cryptosporidium represents a public health risk in terms of the potential for zoonotic transmission.

Age Factors↗

PubMed2004

Complete development of Cryptosporidium parvum in host cell-free culture.

The present study describes the complete in vitro development of Cryptosporidium parvum (cattle genotype) in RPMI-1640 maintenance medium devoid of host cells. This represents the first report in which Cryptosporidium is shown to multiply, develop and complete its life cycle without the need for host cells. Furthermore, cultivation of Cryptosporidium in diphasic medium consisting of a coagulated new born calf serum base overlaid with maintenance medium greatly increased the total number of Cryptosporidium stages. Type I and II meronts were detected giving rise to two morphologically different merozoites. Type I meronts, which appear as grape-like clusters as early as 48 h post culture inoculation, release merozoites, which are actively motile, and circular to oval in shape. Type II meronts group in a rosette-like pattern and could not be detected until day 3 of culturing. Most of the merozoites released from type II meronts are generally spindle-shaped with pointed ends, while others are rounded or pleomorphic. In contrast to type I, merozoites from type II meronts are less active and larger in size. Sexual stages (micro and macrogamonts) were observed within 6-7 days of culturing. Microgamonts were darker than macrogamonts, with developing microgametes, which could be seen accumulating at the periphery. Macrogamonts have a characteristic peripheral nucleus and smooth outer surface. Oocysts at different levels of sporulation were seen 8 days post culture inoculation. Cultures were terminated after 4 months when the C. parvum life cycle was still being perpetuated with the presence of large numbers of excysting and intact oocysts. Culture-derived oocysts obtained after 46 days p.i. were infective to 7- to 8-day-old ARC/Swiss mice. The impact of C. parvum developing in cell-free culture is very significant and will facilitate many aspects of Cryptosporidium research.

Animals↗

PubMed2004

Variation in Giardia: implications for taxonomy and epidemiology.

The taxonomy, life cycle patterns and zoonotic potential of Giardia infecting mammals and birds have been poorly understood and controversial for many years. The development of molecular tools for characterising isolates of Giardia directly from faeces or environmental samples has made an enormous contribution to resolving these issues. It is now clear that the G. duodenalis morphological group is a species complex comprising a series of what appear to be largely host-adapted species, and at least two zoonotic species for which humans are the major host, but which are also capable of infecting other mammals. It is proposed that this new information be reflected in the redesignation of several species of Giardia described previously. The molecular epidemiological tools that are now available need to be applied in different endemic foci of Giardia transmission, as well as in outbreak situations, in order to understand better the frequency of zoonotic transmission as well as to develop more effective approaches to controlling giardiasis.

Animals↗

PubMed2004

Epidemiological and molecular evidence supports the zoonotic transmission of Giardia among humans and dogs living in the same community.

Giardia duodenalis isolates recovered from humans and dogs living in the same locality in a remote tea-growing community of northeast India were characterized at 3 different loci; the SSU-rDNA, elongation factor 1-alpha (ef1-alpha) and triose phosphate isomerase (tpi) gene. Phylogenetic analysis of the SSU-rDNA and efl-alpha genes provided poor genetic resolution of the isolates within various assemblages, stressing the importance of using multiple loci when inferring genotypes to Giardia. Analysis of the tpi gene provided better genetic resolution and placed canine Giardia isolates within the genetic groupings of human isolates (Assemblages A and B). Further evidence for zoonotic transmission was supported by epidemiological data showing a highly significant association between the prevalence of Giardia in humans and presence of a Giardia-positive dog in the same household (odds ratio 3.01, 95% CI, 1.11, 8.39, P = 0.0000).

Adolescent↗

PubMed2004

Cryptosporidium suis n. sp. (Apicomplexa: Cryptosporidiidae) in pigs (Sus scrofa).

Molecular and biological characteristics of a new species of Cryptosporidium from the feces of pigs (Sus scrofa) is described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum; they are passed fully sporulated, lack sporocysts, and measure 4.9-4.4 microm (mean = 4.6 microm) x 4.0-4.3 microm (mean = 4.2 microm); length to width ratio 1.1 (n = 50). Cryptosporidium suis is not transmissible to nude mice and is poorly infectious for cattle. Molecular and phylogenetic analyses at the 18S ribosomal RNA, heat shock protein 70, and actin gene loci demonstrate C. suis to be genetically distinct from all known species and genotypes of Cryptosporidium, and thus is named as Cryptosporidium suis.

Animals↗

PubMed2003

Gastrointestinal parasites of domestic cats in Perth, Western Australia.

A study was conducted to determine the prevalence of gastrointestinal parasites in a sample of domestic cats in Perth and the knowledge of their owners about the control and potential for zoonotic transmission of these parasites. Faecal samples (418), collected from cats originating from five sources, were examined by microscopy and questionnaires administered to cat owners. Forty randomly selected samples were also screened using PCR in order to detect cysts of Giardia and oocysts of Cryptosporidium that may have been present in a faecal sample at very low levels. The overall prevalence of gastrointestinal parasites in domestic cats by microscopy was 8.6%. Pet shop kittens had the highest parasite prevalence (34.3%), followed by cats and kittens from breeding establishments (15.8%), refuge cats and kittens (8.3%), privately owned cats (2.3%), and boarding cats and kittens (1.6%). Surprisingly, 80% of the 40 cats tested by PCR were positive for Giardia duodenalis and 10% for Cryptosporidium. None of these cats were positive on microscopy. After adjusting for other factors with multiple logistic regression, kittens less than 6 months of age, and cats living in households with more than one cat or with a dog were significantly more likely to be parasitised. In the logistic regression model, the presence of parasitism was also significantly influenced by the number of anthelmintic doses administered in the 12 month period prior to the study. The majority (64.5%) of cat owners were aware that feline parasites could be transmitted to humans, however less than half (42.8%) were aware of the modes of transmission to humans.

Age Factors↗

PubMed2003

Detection of the rodent tapeworm Rodentolepis (=Hymenolepis) microstoma in humans. A new zoonosis?

A longitudinal survey of gastro-intestinal parasites was conducted over a 3-year period in remote communities in the north-west of Western Australia where, based on diagnosis by microscopy of faecal samples, Rodentolepis (=Hymenolepis) nana was found to be the most common enteric parasite. In the present study, using molecular tools, we describe the unexpected discovery, of a mixed infection with a second hymenolepidid species, Rodentolepis (=Hymenolepis) microstoma in four of the surveyed individuals. In the absence of any reliable earlier reports we believe this is to be the first instance of the detection of R. microstoma from human hosts. The development of a diagnostic restriction fragment polymorphism has enabled the study of R. microstoma in human populations and will greatly facilitate a more thorough understanding of the epidemiology of this parasite in the future.

Animals↗

PubMed2003

Humans, dogs and parasitic zoonoses--unravelling the relationships in a remote endemic community in northeast India using molecular tools.

Canine parasitic zoonoses pose a continuing public health problem, especially in developing countries and communities that are socioeconomically disadvantaged. Our study combined the use of conventional and molecular epidemiological tools to determine the role of dogs in transmission of gastrointestinal (GI) parasites such as hookworms, Giardiaand Ascarisin a parasite endemic tea-growing community in northeast India. A highly sensitive and specific PCR-RFLP was developed to detect and differentiate the zoonotic species of canine hookworm eggs directly from faeces. This allowed epidemiological screening of canine hookworm species in this community to be conducted with ease and accuracy. Seventy two percent of dogs were found to harbour A. caninum, 60% A. braziliense and 37% harboured mixed infections with both hookworms. No A. ceylanicum was detected in the dog population. The zoonotic potential of canine Giardiawas also investigated by characterising Giardia duodenalisrecovered from humans and dogs living in the same locality and households, at three different loci. Phylogenetic and epidemiological analysis provided compelling evidence to support the zoonotic transmission of canine Giardia. Molecular tools were also used to identify the species of Ascarisegg present in over 30% of dog faecal samples. The results demonstrated the role of dogs as a significant disseminator and environmental contaminator of Ascaris lumbricoidesin communities where promiscuous defecation practices exist. Our study demonstrated the usefulness of combining conventional and molecular parasitological and epidemiological tools to help solve unresolved relationships with regards to parasitic zoonoses.

Ancylostomatoidea↗