Loss of Keratin 17 induces tissue-specific cytokine polarization and cellular differentiation in HPV16-driven cervical tumorigenesis in vivo.

Despite preventive human papilloma virus (HPV) vaccination efforts, cervical cancer remains a leading cause of death in women worldwide. Development of therapeutic approaches for cervical cancer are hampered by a lack of mechanistic insight during tumorigenesis. The cytoskeletal protein Keratin 17 (KRT17;K17) is robustly expressed in a broad array of carcinomas, including in cervical tumors, where it has both diagnostic and prognostic value. In this study, we have established multiple functional roles for K17 in the promotion of cervical tumorigenesis in vivo using the established HPV16tg mouse model for cervical squamous cell carcinoma. In HPV16tg/+;Krt17-/-relative to HPV16tg/+ reference female mice, onset of cervical lesions is delayed and closely paralleled by marked reductions in hyperplasia, dysplasia and vascularization. In addition, loss of Krt17 is associated with a cytokine polarization and recruitment of effector immune cells to lesion-prone cervical epithelia. Further, we observed marked enhancement of terminal differentiation in HPV16tg/+;Krt17-/-cervical epithelium accompanied by a stimulation and expansion in the expression of p63, a known basal/reserve cell marker in this tissue. Altogether, the data suggest that the loss of Krt17 may foster an overall protective environment for lesion-prone cervical tissue. In addition to providing new insights into the immunomodulatory and cellular mechanisms of cervical tumorigenesis, these findings may help guide the development of future therapies including vaccines.

A new Eimeria species parasitic in Isoodon obesulus (Marsupialia: Peramelidae) in Western Australia.

Feces from southern brown bandicoots, Isoodon obesulus, inhabiting the Perth metropolitan area were examined using fecal flotation and light microscopy, and were frequently found to contain oocysts (10/24; 42%). To enable formal description of the proposed new Eimeria species, i.e., Eimeria quenda n. sp., fecal oocysts from 1 juvenile male I. obesulus were allowed to sporulate in 2% potassium dichromate (K(2)Cr(2)O(7)) at room temperature. Sporulated oocysts are spheroidal to subspheroidal 24.5 × 23.6 (22.5-26.0 × 22.5-24.8) µm, with L/W ratio of 1.04 (1.00-1.13), lack a micropyle and oocyst residuum, and are contained within a smooth trilaminate oocyst wall 1.8 (1.6-2.0) µm thick. Sporocysts are ovoid, 12.6 × 9.2 (12.0-13.8 × 8.5-10.0) µm, with L/W ratio of 1.37 (1.20-1.53), have a sporocyst residuum, and 2 comma-shaped sporozoites, each containing 2 spheroidal refractile bodies. Sporulation takes 1-3 days at room temperature. This is the second formal description of an Eimeria species parasitic in the order Peramelemorphia.

Echinococcus granulosus from Mexican pigs is the same strain as that in Polish pigs.

Samples of Echinococcus granulosus from seven pigs from Mexico were compared with isolates of the parasite from pigs in Poland and representative strains and species of Echinococcus. Isolates from pigs in Mexico were found to be genetically identical to E. granulosus from Polish pigs and distinct from other major genotypes by sequencing part of the mitochondrial cytochrome c oxidase I (COI) mtDNA locus, restriction fragment length polymorphism (RFLP) of the polymerase chain reaction (PCR) amplified rDNA internal transcribed spacer (ITS) 1 using five different enzymes, and random amplified polymorphic DNA (RAPD) analysis. These results were complemented by data on hook morphology and together strengthen the view that Echinococcus maintained in a cycle involving pigs and dogs is a distinct strain that is conserved genetically in different geographical areas. The present study supports the close relationship of the cervid, camel and pig strains and raises the question of their taxonomic status.

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.

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.

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.

A new eimeria species parasitic in western barred bandicoots, Perameles bougainville (Marsupialia: Peramelidae), in western Australia.

Feces from western barred bandicoots, Perameles bougainville, examined during routine monitoring of captive breeding colonies and wild populations were frequently found to contain oocysts. Fecal oocysts from 1 individual housed at Kanyana Wildlife Rehabilitation Centre were allowed to sporulate in 2% potassium dichromate (K2Cr2O7) at room temperature. Sporulated oocysts are subspheroidal 18.8 X 17.9 (16.9-21.0 x 16.0-19.9) microm, with length/width (L/W) ratio of 1.05 (1.00-1.15), lack a micropyle and oocyst residuum, but they usually have a polar granule within a smooth trilaminate oocyst wall 1.0 (0.7-1.3) microm thick. Sporocysts are ovoid, 9.1 x 7.0 (8.1-10.8 x 6.1-8.6) microm, with L/W ratio of 1.32 (1.04-1.51), have a Stieda body, sporocyst residuum, and 2 comma-shaped sporozoites, each containing 2 spheroidal refractile bodies. Sporulation takes 2-5 days at room temperature. This is the first formal description of an Eimeria species parasitic in the order Peramelemorphia.

Occurrence of gastrointestinal parasites in horses in metropolitan Perth, Western Australia.

OBJECTIVE: To assess the occurrence of gastrointestinal parasites in horses in Perth. To apply polymerase chain reaction (PCR) for the identification of some species of encysted larval cyathostomes. DESIGN: Between February and September of 2000, the gastrointestinal tracts of 29 horses submitted to a local knackery and Murdoch University Veterinary hospital in Perth were examined post mortem for the presence of gastrointestinal parasites. PROCEDURE: The gastrointestinal tract was divided into six sections, which were screened for the presence of parasites such as Gasterophilus sp, Anoplocephala sp and Parascaris equorum. Samples of contents were taken for worm counts. RESULTS: Cyathostomes were found in 28 of the 29 horses. Eighteen species of gastrointestinal helminths were identified. Twelve of these were cyathostomes, with the four most common species being Cyathostomum catinatum, Cylicocyclus nassatus, Cylicostephanus longibursatus and Cylicostephanus goldi. The large strongyle, Triodontophorus serratus, was found in three of the horses but species of Strongylus were not found. CONCLUSIONS: In contrast to a study conducted on horses from this region in 1985, cyathostomes were the most common gastrointestinal parasites found. The four most common cyathostome species found in the present study correlated well with findings of studies in other locations. The high number of cyathostomes may be due to the increase in resistance to anthelmintics among the species, and to improper anthelmintic use. The apparent reduction in number of large strongyles may be due to the widespread use of ivermectin, which is very effective against these parasites, and also possibly because some larvae may not have been detected.

Larval anisakid infections of some tropical fish species from north-west Australia.

Despite the commercial and zoonotic importance of larval anisakid infestations of teleosts, their distribution among Australia's diverse marine fish fauna is poorly understood. A preliminary survey of Australia's tropical north-west revealed a generally high prevalence of larval anisakids representing four genera (Anisakis, Terranova, Thynnascaris and Raphidascaris) among only seven fish species. The potential impact of high larval anisakid infections on both the health of recreational fishermen and aquaculture environments is discussed.

Pathogens of house mice on arid Boullanger Island and subantarctic Macquarie Island, Australia.

Studies on island populations of house mice (Mus domesticus) and their viruses reveal insights into viral persistence in isolated communities. We surveyed the ectoparasites, endoparasites, and antiviral antibodies for 11 murine viruses and two bacteria of house mice inhabiting two islands off Australia. House mice on Boullanger Island were seropositive to two viruses, murine cytomegalovirus and epizootic diarrhea of infant mice. On subantarctic Macquarie Island, house mice were seropositive for five viruses: murine cytomegalovirus, lymphocytic choriomeningitis virus, mouse parvovirus, epizootic diarrhea of infant mice, and Theiler's murine encephalomyelitis virus. The diversity of antiviral antibodies was lower among populations of house mice on islands than those inhabiting mainland Australia. The decreased diversity of viruses in island populations of house mice may be a function of which agent the founder mice transfer to the island and related to the low densities which the host population may periodically reach over time.

Teaching of parasitology to students of veterinary medicine and biomedical sciences.

The teaching of an applied parasitology course suitable for both veterinary and biomedical students is described. A common lecture course is given complemented by separate and specific practical, research and problem-based learning components designed for veterinary and biomedical students. For veterinary and biomedical students, teaching of parasitology during the full course comprises a total of 46 lectures; 13 practical classes for veterinary students and five for biomedical students who also undertake an independent research project.

Molecular and morphological characterization of Echinococcus granulosus of human and animal origin in Iran.

Iran is an important endemic focus of cystic hydatid disease (CHD) where several species of intermediate host are commonly infected with Echinococcus granulosus. Isolates of E. granulosus were collected from humans and other animals from different geographical areas of Iran and characterized using both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). The sheep and camel strains/genotypes were shown to occur in Iran. The sheep strain was shown to be the most common genotype of E. granulosus affecting sheep, cattle, goats and occasionally camels. The majority of camels were infected with the camel genotype as were 3 of 33 human cases. This is the first time that cases of CHD in humans have been identified in an area where a transmission cycle for the camel genotype exists. In addition, the camel genotype was found to cause infection in both sheep and cattle. Results also demonstrated that both sheep and camel strains can be readily differentiated on the basis of hook morphology alone.

Factors influencing the fecal egg and oocyst counts of parasites of wild European rabbits Oryctolagus cuniculus (L.) in Southern Western Australia.

Abundance of intestinal parasites was monitored by fecal egg and oocyst counts for samples of wild rabbits Oryctolagus cuniculus with different levels of imposed female sterility from 12 populations in southwestern Australia. Differences in egg counts of Trichostrongylus retortaeformis between seasons and age groups were dependent on the sex of the host. Pregnancy may have been responsible for these differences because egg counts were consistently higher in intact females than in females surgically sterilized by tubal ligation. Egg counts for Passalurus ambiguus were influenced by season and host age but there were no differences between sexes or between intact and sterilized female rabbits. No differences were detected in the oocyst counts of the 8 species of Eimeria between male and female rabbits or between intact and sterilized females. Seasonal differences were detected in oocyst counts of Eimeria flavescens and Eimeria stiedai. The overwhelming determinant of coccidian oocyst counts was host age, with 6 species being much more abundant in rabbits up to 4 mo of age. There was a suggestion that egg counts of T. retortaeformis and oocyst counts of several species of Eimeria were reduced in populations where rabbit numbers had been depressed for at least 2 yr, but there was no evidence that short-term variations in rabbit numbers had a measurable effect on parasite abundance.

Evaluation of the association of parasitism with mortality of wild European rabbits Oryctolagus cuniculus (L.) in southwestern Australia.

Abundances of the parasitic nematodes Trichostrongylus retortaeformis and Passalurus ambiguus, and 8 Eimeria species were estimated by fecal egg and oocyst output in 12 discrete free-ranging populations of wild rabbits (Oryctolagus cuniculus) in southwestern Australia. Comparisons of parasite egg and oocyst counts were made between those rabbits known to have survived at least 2 mo after fecal samples were collected and those rabbits that did not survive. There were significant negative relationships between parasite egg and oocyst counts and survival when all age groups and collection periods were pooled for several species of coccidia and for T. retortaeformis. However, when the same comparisons were made within rabbit age groups and within collection periods, there were very few significant differences even where sample sizes were quite large. The differences indicated by the pooled analysis for coccidia were most likely due to an uneven host age distribution with respect to survival, combined with an uneven distribution of the oocyst counts with rabbit age. The result for T. retortaeformis was similarly affected but by a seasonal pattern. Parasitism by nematodes and coccidia did not appear to be an important mortality factor in these rabbit populations, at least at the range of host densities we examined. This suggests that other factors must have been responsible for the observed pattern of density-dependent regulation in these rabbits.

Efficacy of albendazole against Giardia and hookworm in a remote Aboriginal community in the north of Western Australia.

The parasitological, clinical efficacy and tolerability of albendazole in the treatment for both giardiasis and hookworm infection in a remote Aboriginal population was investigated. Albendazole at a dose rate of 400 mg daily for 5 days was highly effective in reducing hookworm egg numbers and both Giardia antigen and cysts. The 36.6% prevalence of Giardia prior to treatment fell to 12% between days 6 and 9, 15% for days 10-17 and rose to 28% between days 18 and 30. Tolerability and clinical efficacy were excellent. The effect of albendazole on hookworm was longer lasting than that on Giardia, reducing percent infection from over 76-2% on days 6-9 and zero by day 18-30 despite conditions highly conducive to rapid re-infection. We conclude that albendazole is highly efficacious against both parasites when used as described but that long term community benefit may require additional education programmes to avoid re-infection with Giardia although treatment strategies would seem appropriate for hookworm.

Platynosomum fastosum in ex-captive orangutans from Indonesia.

The liver fluke Platynosomum fastosum was identified upon necropsy of three ex-captive orangutans (Pongo pygmaeus) which had been part of a rehabilitation program for reintroduction to the wild. This trematode has not been reported in orangutans previously and is commonly found in cats in Southeast Asia. Cross infection from cats via intermediate hosts, to orangutans kept in captivity as pets, could explain their presence in the latter. Although P. fastosum caused intrahepatic and bile duct damage, death of the hosts could not be attributed solely to the presence of the liver fluke infection.

Gram-negative bacterial infections and cardiovascular parasitism in green sea turtles (Chelonia mydas).

OBJECTIVE: To investigate causes of ill health and mortality in juvenile wild green sea turtles (Chelonia mydas) found along the mid-north west coast of Western Australia between June and October of 1997. PROCEDURE: Department of Conservation and Land Management rangers submitted four dead or dying green sea turtles from separate incidents for veterinary examination, necropsy, and bacteriological, parasitological and histopathological examination. RESULTS: Numerous different species of trematodes belonging to the families Pronocephalidae, Microscaphidiidae and Paramphistomidae were detected in the intestines of two turtles examined, and in all turtles there was severe spirorchid fluke infection including Haemoxenicon sp, Amphiorchis sp and Hapalotrema sp. Histopathological examination demonstrated severe multifocal to diffuse granulomatous vasculitis, aggregations of spirorchid fluke eggs and microabscesses throughout various tissues including intestines, kidney, liver, lung and brain. Cultures and or histopathological examination demonstrated disseminated Gram-negative bacterial infections including salmonella, E coli, Citrobacter freundii and Moraxella sp. CONCLUSION: Infections caused by salmonellae, E coli and other Gram-negative bacteria should be considered as causes of systemic illness and death in wild green sea turtles infected with spirorchid cardiovascular flukes and other internal parasites.

The prevalence of hookworm infection, iron deficiency and anaemia in an aboriginal community in north-west Australia.

OBJECTIVE: To determine the prevalence of hookworm infections, iron deficiency and anaemia in an Aboriginal community in the north of Western Australia. DESIGN AND SETTING: A cross-sectional survey conducted in 1992, examining faecal specimens and blood samples from Aboriginals and non-Aboriginals in a remote coastal community in the north of Western Australia. PARTICIPANTS: All those living in the community at the time of the survey. MAIN OUTCOME MEASURES: Parasite status and haematological values for haemoglobin, serum iron, ferritin, transferrin and mean red cell volume. RESULTS: Infections with hookworm were present throughout the Aboriginal population (77%; n = 243), with the highest prevalence in children aged 5-14 years (93%; n = 74). Hookworm was not detected in non-Aboriginals (n = 24). Iron deficiency was common throughout the Aboriginal population, especially in children aged 5-14 years (79%; n = 68) and women aged over 14 years (72%; n = 65). Anaemia was highly prevalent among Aboriginal children aged 5-14 years (84% in hookworm-positive children, 75% in hookworm-negative) and women aged over 14 years (63% in hookworm-positive women, 31% in hookworm-negative). Aboriginals over 14 years of age who had hookworm (n = 82) had significantly lower levels of haemoglobin, serum iron and serum ferritin, a lower mean red cell volume and significantly higher transferrin levels than uninfected Aboriginals (n = 38) and non-Aboriginals (n = 19) in the same age group. Hookworm infections were associated with anaemia (P < 0.01) and iron deficiency (P < 0.01) in people over 14 years of age. The species of hookworm, determined after examining 13 larval cultures and two adult worms, was found to be Ancylostoma duodenale. CONCLUSION: Infections with A. duodenale are endemic in Aboriginals in this community, and are likely to contribute to the high prevalence of iron deficiency and anaemia observed in the Aboriginal population, particularly in children and women.