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1.
Aust Vet J ; 102(1-2): 35-40, 2024.
Article in English | MEDLINE | ID: mdl-38057960

ABSTRACT

Strongyloides stercoralis, the causative agent of strongyloidiasis, is a potentially zoonotic intestinal nematode endemic to northern Australia. Strongyloidiasis is typically observed in immunocompromised hosts and is characterised by gastrointestinal signs, respiratory symptoms and a failure to thrive. In immunocompromised hosts, hyperinfection syndrome and disseminated infections can prove life-threatening. A 24-month-old Boston Terrier dog was referred for investigation of chronic small and large intestinal watery hematochezic diarrhoea, emaciation and hematemesis. Small intestinal histology identified a nematode despite consecutive negative faecal flotations. A real-time polymerase chain reaction and Baermann test subsequently confirmed infection with S. stercoralis. The dog had received an oral parasiticide comprising milbemycin oxime and afoxolaner every month for the 11 months prior to this diagnosis. Despite fenbendazole being reported as successful in the treatment of canine strongyloidiasis, a course of fenbendazole failed to clear the infection. Eradication of S. stercoralis infection was confirmed after the administration of off-label ivermectin fortnightly for 12 doses. Attention should be paid to this nematode as the failure of routine copromicroscopic methods to diagnose S. stercoralis infections can result in misdiagnosis, mistreatment and progression of the disease. Off-label ivermectin may be an alternative to fenbendazole for the treatment of Strongyloides spp. infection in dogs.


Subject(s)
Dog Diseases , Strongyloides stercoralis , Strongyloidiasis , Dogs , Animals , Strongyloidiasis/diagnosis , Strongyloidiasis/drug therapy , Strongyloidiasis/veterinary , Ivermectin/therapeutic use , Fenbendazole/therapeutic use , Feces , Dog Diseases/diagnosis , Dog Diseases/drug therapy , Dog Diseases/epidemiology
2.
J Helminthol ; 94: e43, 2019 Feb 28.
Article in English | MEDLINE | ID: mdl-30813972

ABSTRACT

All canine hookworms are known to be zoonotic, causing infections ranging from transient skin irritations to prolonged 'creeping eruptions', eosinophilic enteritis and even patent intestinal infections. There is little information on canine hookworm species and their public health significance in sub-Saharan Africa. This study determined the prevalence and species of hookworms in dogs from different climatic zones of Kenya. Dog faecal samples were collected from the environment, and hookworm eggs were isolated by zinc chloride flotation and subjected to DNA extraction. Polymerase chain reaction (PCR) assays targeting the internal transcribed spacer (ITS) 1 and 2, 5.8S and 28S ribosomal RNA of Ancylostoma spp. and Uncinaria stenocephala were performed, and hookworm species were identified by PCR-restriction fragment length polymorphism (RFLP) or DNA sequencing. Hookworm eggs were detected by microscopy in 490/1621 (30.23%, 95% CI 28.01-32.54) faecal samples. Estimates of faecal prevalence were high in counties receiving higher rainfall (Narok 46.80%, Meru 44.88%) and low in those with a more arid climate (Isiolo 19.73%, Turkana 11.83%). In a subset of 70 faecal samples, Ancylostoma caninum (n = 59) was the most common species, followed by A. braziliense (n = 10) and A. cf. duodenale (n = 1). This study reports for the first time the detection of A. cf. duodenale in dog faeces and zoonotic hookworm species in Kenyan dogs. These findings emphasize the need for control measures such as enforcing laws for restraining stray dogs, regular deworming of dogs, and public health awareness programmes aimed at informing communities on outdoor use of footwear.


Subject(s)
Ancylostomatoidea/isolation & purification , Dog Diseases/parasitology , Hookworm Infections/veterinary , Ancylostomatoidea/classification , Ancylostomatoidea/genetics , Animals , Dogs , Feces/parasitology , Female , Hookworm Infections/parasitology , Kenya , Male , Polymorphism, Restriction Fragment Length
3.
Article in English | MEDLINE | ID: mdl-28730748

ABSTRACT

BACKGROUND: We previously developed an animal model to examine mechanisms that underlie the emergence of visceral hypersensitivity modeling pain characteristics of temporomandibular disorder (TMD) patients with comorbid irritable bowel syndrome (IBS). In ovariectomized (OVx) rats with estradiol (E2) replacement, visceral hypersensitivity developed subsequent to masseter muscle inflammation followed by repeated forced swim (FS) stress. The purpose of this study was to investigate whether activation of extracellular signal-regulated kinase (ERK) in the spinal cord contributes to visceral hypersensitivity in this overlapping pain model. METHODS: In OVx with E2 replacement rats masseter muscle inflammation was followed by 3 day FS (comorbid condition). Depression-like behaviors were assessed by sucrose preference and in the elevated plus maze, and visceral sensitivity was measured by the visceromotor response (VMR) to colorectal distention. The protein level of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) in the L6-S2 dorsal spinal cord was analyzed by western blot. KEY RESULTS: FS stress decreased sucrose consumption in E2 replaced rats in sucrose preference test. The expression of p-ERK1/2 in the L6-S2 dorsal spinal cord increased significantly in E2 with comorbid rats. Intrathecal injection of mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor PD98059 blocked the visceral hypersensitivity induced by masseter muscle inflammation combined with FS stress. CONCLUSIONS & INFERENCES: These data indicate that ERK1/2 activation contributes to the visceral hypersensitivity evoked by craniofacial inflammation pain combined with stress. The results may provide a new therapeutic avenue for alleviating overlapping pain conditions.


Subject(s)
Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Myositis/metabolism , Spinal Cord/metabolism , Stress, Psychological/metabolism , Visceral Pain/metabolism , Animals , Depression/etiology , Estradiol/administration & dosage , Female , Masseter Muscle/physiopathology , Myositis/complications , Ovariectomy , Phosphorylation , Rats, Sprague-Dawley , Stress, Psychological/complications , Visceral Pain/complications
4.
Neurogastroenterol Motil ; 27(6): 775-86, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25810326

ABSTRACT

BACKGROUND: We previously reported estrogen modulates spinal N-methyl-d-aspartate (NMDA) receptor processing of colorectal pain through changes in spinal GluN1 subunit phosphorylation/expression. The purpose of this study was to investigate whether spinal GluN2B containing NMDA receptors are involved in estrogen modulation of visceral pain processing. METHODS: Behavioral, molecular, and immunocytochemical techniques were used to determine spinal GluN2B expression/phosphorylation and function 48 h following subcutaneous injection of estradiol (E2) or vehicle (safflower oil, Saff oil) in ovariectomized rats in the absence or presence of colonic inflammation induced by mustard oil. KEY RESULTS: E2 increased the magnitude of the visceromotor response (VMR) to colorectal distention compared to Saff oil in non-inflamed rats. Intrathecal injection of the GluN2B subunit antagonist, Ro 25-6981, had no effect on the VMR in non-inflamed E2 or Saff oil rats. Colonic inflammation induced visceral hyperalgesia in E2, but not Saff oil rats. Visceral hyperalgesia in E2 rats was blocked by intrathecal GluN2B subunit selective antagonists. In inflamed rats, E2 increased GluN2B protein and gene expression in the thoracolumbar (TL), but not lumbosacral (LS), dorsal spinal cord. Immunocytochemical labeling showed a significant increase in GluN2B subunit in the superficial dorsal horn of E2 rats compared to Saff oil rats. CONCLUSIONS & INFERENCES: These data support the hypothesis that estrogen increases spinal processing of colonic inflammation-induced visceral hyperalgesia by increasing NMDA receptor activity. Specifically, an increase in the activity of GluN2B containing NMDA receptors in the TL spinal cord by estrogen underlies visceral hypersensitivity in the presence of colonic inflammation.


Subject(s)
Behavior, Animal , Colitis/metabolism , Estradiol/pharmacology , Estrogens/pharmacology , Hyperalgesia/metabolism , Receptors, N-Methyl-D-Aspartate/drug effects , Spinal Cord/drug effects , Visceral Pain/metabolism , Animals , Colitis/chemically induced , Colitis/complications , Female , Hyperalgesia/etiology , Immunohistochemistry , Lumbar Vertebrae , Mustard Plant , Ovariectomy , Phenols/pharmacology , Phosphorylation , Piperidines/pharmacology , Plant Oils , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Receptors, N-Methyl-D-Aspartate/metabolism , Spinal Cord/metabolism , Thoracic Vertebrae , Visceral Pain/etiology
5.
Aust Vet J ; 93(3): 58-66, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25708788

ABSTRACT

OBJECTIVES: To estimate the proportion of canine tick-borne disease (CTBD) pathogens in dogs from northern states of Australia presenting with and without clinical signs/laboratory abnormalities suggestive of CTBD and to evaluate associated risk factors. DESIGN: Client-owned dogs presented to a general practice clinic in the Northern Territory (NT; n = 138) and five referral hospitals in south-east Queensland (SEQ; n = 100) were grouped into CTBD-suspect and -control groups based on clinical and laboratory criteria. Blood and sera were screened for haemotropic Mycoplasma spp., Babesia spp., Anaplasma spp., Ehrlichia spp. and Hepatozoon spp. using microscopic examination, in-clinic ELISA testing and PCR assays. Dog-specific risk factors associated with the presence of CTBD pathogens were evaluated. RESULTS: Overall, 24.4% of the suspect group and 12.2% of the control group dogs were infected. The proportions of M. haemocanis, B. vogeli, A. platys, Candidatus Mycoplasma haematoparvum, and C. Mycoplasma haemobos were 7.1%, 5.0%, 3.8%, 1.7% and 0.4%, respectively. Dogs originating from the NT were 3.6-fold (95% confidence interval (CI) 1.51-8.62; P = 0.004) more likely to be infected with CTBD pathogens than those from SEQ. Male dogs were 2.3-fold (95% CI 1.17-4.80, P = 0.024) more likely to be PCR-positive to CTBD pathogens than female dogs. Dogs presenting with clinical signs consistent with CTBD and thrombocytopenia were more likely to be infected by CTBD pathogens (odds ratio 2.85; 95% CI 1.16, 7.02; P = 0.019). CONCLUSIONS: Haemotropic mycoplasmas were the most common tick-borne pathogen infecting client-owned dogs. Subclinical cases were common in dogs from the NT. Veterinary practitioners should be aware of the proportion of CTBD pathogens and the presenting features of clinical and subclinical disease in their area.


Subject(s)
Dog Diseases/parasitology , Tick-Borne Diseases/veterinary , Anaplasma , Anaplasmosis/etiology , Anaplasmosis/transmission , Animals , Babesia , Babesiosis/etiology , Babesiosis/transmission , Dog Diseases/etiology , Dogs/parasitology , Ehrlichia canis , Ehrlichiosis/etiology , Ehrlichiosis/transmission , Ehrlichiosis/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Male , Mycoplasma , Mycoplasma Infections/etiology , Mycoplasma Infections/transmission , Mycoplasma Infections/veterinary , Northern Territory , Polymerase Chain Reaction/veterinary , Queensland , Risk Factors , Sex Factors , Tick-Borne Diseases/etiology , Tick-Borne Diseases/parasitology
6.
Parasite Immunol ; 36(10): 503-8, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24920168

ABSTRACT

Blastocystis is an intestinal protist found in many species including humans and pigs. It has a controversial pathogenesis and has been implicated as a potential cause of irritable bowel syndrome. Our previous studies identified pigs as potential animal models for blastocystosis by demonstrating that they were likely natural hosts of Blastocystis and can harbour subtypes (ST) in common with humans. Furthermore, our finding of a lack of intestinal histopathology associated with Blastocystis infection in pigs is also a consistent finding in examined infected humans. In this study, we aimed to identify and characterize the Blastocystis-specific mucosal IgA response in pigs by immunoblotting, using pig faecal antibodies and Blastocystis antigen. Faeces from 233 pigs representing three age groups (sows/boars, growers/weaners and piglets) and including five dexamethasone-immunosuppressed research pigs were tested. The majority (81·5%) of the pigs had faecal IgA reactivity against Blastocystis proteins of molecular weights of 17·5-120 kDa. Reactivity to a >250 kDa protein was found in 18·5% of pigs. Notably, immunosuppressed pigs and piglets were statistically more likely to have reactivity to this protein compared to growers/weaners and sows/boars, respectively. These results corroborate other findings suggesting that compromised immunity may predispose to blastocystosis and support our contention that pigs are potentially good models for pathogenesis studies.


Subject(s)
Blastocystis Infections/veterinary , Blastocystis/isolation & purification , Immunoglobulin A/immunology , Swine Diseases/immunology , Animals , Blastocystis Infections/immunology , Blastocystis Infections/parasitology , Feces/parasitology , Female , Male , Swine , Swine Diseases/parasitology
7.
Aust Vet J ; 91(3): 89-93, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23438459

ABSTRACT

OBJECTIVE: To investigate the cause of an outbreak of bovine cysticercosis (Taenia saginata) infection on a cattle property in north-western New South Wales (NSW). METHODS: Cystic lesions were detected in the muscles of cattle during routine meat inspection at slaughter. These lesions were confirmed to be cysticerci of T. saginata through histology and polymerase chain reaction (PCR). Data on cattle maintenance were obtained through interviews with feedlot owners and staff. A suspect feed supplement was investigated. RESULTS: Between 5 July to 13 December 2010, 390 feedlot cattle from north-western NSW were slaughtered in abattoirs in NSW and Queensland. Of these, 138 animals had been maintained exclusively in feedlot enclosures from 80 to 300 days. Bovine cysticercosis was discovered in 80 cattle (58%, 26 carcasses were condemned). Another 18 cattle spent 24 h in the feedlot before being moved onto pasture and 1 of them was found to be infected. During the 5 months following the initial outbreak, a further 275 cattle were slaughtered; 2 of 51 heifers retained in the feedlot for a further 100 days were infected. None of the 234 animals grazed exclusively on pasture on the property were infected. Bovine cysticercosis was confirmed through examination of histological sections of muscle lesions and PCR using DNA extracted from cysticerci. No eggs of T. saginata were recovered from the feed supplement using a standard flotation method. CONCLUSIONS: The source of infection arose from rations contaminated with human faeces. All possibilities for local contamination were eliminated during the investigation. The suspected source of infection was imported copra meal, which was used as a feed supplement.


Subject(s)
Cattle Diseases/epidemiology , Cysticercosis/veterinary , Food Contamination/analysis , Abattoirs , Animal Feed , Animals , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/etiology , Cysticercosis/diagnosis , Cysticercosis/epidemiology , Cysticercosis/etiology , Disease Outbreaks , Female , Food Parasitology , Male , New South Wales/epidemiology
8.
Aust Vet J ; 90(4): 130-5, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22443327

ABSTRACT

OBJECTIVES: To determine the prevalence of canine vector-borne diseases (CVBD: Babesia spp., Anaplasma spp., Ehrlichia spp., haemotropic mycoplasmas and Hepatozoon) in Australian dogs; namely, dogs from pounds in south-east Queensland and an indigenous Aboriginal community in the north-east of the Northern Territory. DESIGN AND PROCEDURE: Blood samples were collected from 100 pound dogs and 130 Aboriginal community dogs and screened for the CVBD pathogens using polymerase chain reaction (PCR). All positive PCR products were sequenced for species confirmation. RESULTS: In total, 3 pound dogs and 64 Aboriginal community dogs were infected with at least one CVBD pathogen. Overall, B. vogeli was detected in 13 dogs, A. platys in 49, M. haemocanis in 23, Candidatus Mycoplasma haematoparvum in 3 and C. M. haemobos in 1 dog. Co-infections were detected in 22 Aboriginal community dogs. CONCLUSIONS: This study found B. vogeli, A. platys and haemotropic mycoplasma infections to be common in dogs in subtropical and tropical areas of Australia. This study also reports for the first time the prevalence and genetic characterisation of haemotropic mycoplasmas in dogs in Australia.


Subject(s)
Dog Diseases/epidemiology , Tick-Borne Diseases/veterinary , Anaplasma/isolation & purification , Anaplasma/pathogenicity , Animals , Babesia/isolation & purification , Babesia/pathogenicity , Dog Diseases/microbiology , Dog Diseases/parasitology , Dogs , Ehrlichia canis/isolation & purification , Ehrlichia canis/pathogenicity , Female , Male , Northern Territory/epidemiology , Polymerase Chain Reaction/veterinary , Protozoan Infections, Animal/epidemiology , Queensland/epidemiology , Tick-Borne Diseases/epidemiology , Tick-Borne Diseases/microbiology , Tick-Borne Diseases/parasitology
9.
Vet Parasitol ; 185(2-4): 343-5, 2012 Apr 30.
Article in English | MEDLINE | ID: mdl-22030374

ABSTRACT

Multiple Trichinella species are reported from the Australasian region although mainland Australia has never confirmed an indigenous case of Trichinella infection in humans or animals. Wildlife surveys in high-risk regions are essential to truly determine the presence or absence of Trichinella, but in mainland Australia are largely lacking. In this study, a survey was conducted in wild pigs from mainland Australia's Cape York Peninsula and Torres Strait region for the presence of Trichinella, given the proximity of a Trichinella papuae reservoir in nearby PNG. We report the detection of a Trichinella infection in a pig from an Australian island in the Torres Strait, a narrow waterway that separates the islands of New Guinea and continental Australia. The larvae were characterised as T. papuae (Kikori strain) by PCR and sequence analysis. No Trichinella parasites were found in any pigs from the Cape York Peninsula. These results highlight the link the Torres Strait may play in providing a passage for introduction of Trichinella parasites from the Australasian region to the Australian mainland.


Subject(s)
Sus scrofa , Trichinella/isolation & purification , Trichinellosis/veterinary , Animals , Australia/epidemiology , Geography , Trichinellosis/epidemiology
10.
Intern Med J ; 42(11): 1187-95, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22032439

ABSTRACT

BACKGROUND: Blastocystis is a common, enteric parasite. The pathogenicity of the organism is uncertain, but subtypes (ST) 1 and 3 have been reported more likely to cause irritable bowel-like symptoms. AIMS: We treated symptomatic patients positive for Blastocystis with conventional therapy and analysed 16 small-subunit (SSU) rDNA to assess clearance and carriage rates and ST prevalence of the parasite in the asymptomatic household members. METHODS: In a longitudinal, prospective case study, 11 symptomatic patients positive for Blastocystis underwent outpatient clinical assessment to exclude other diagnoses before 14 days of either metronidazole 400 mg three times daily or trimethoprim/sulfamethoxazole 160/800 mg twice-daily therapy. Faecal specimens were collected from patients at baseline, day 15, 28 and 56 after therapy and from 17 family members and eight pets at day 15. Specimens were analysed using faecal smear, culture and polymerase chain reaction analysis of 16SSU rDNA. RESULTS: No patient cleared the organism following therapy. ST 1 (45%), 3 (36%), 4 (36%) and 6 (9%) were found in the symptomatic Blastocystis patients, and ST identified before and after therapy were identical in each individual. All household contacts were positive for Blastocystis and 16/17 (94%) contacts showed identical Blastocystis ST to the symptomatic family member. All pets were positive for Blastocystis with polymerase chain reaction testing, 7/8 (88%) demonstrating ST concordance with the symptomatic Blastocystis patients. CONCLUSIONS: Conventional therapy is ineffective for symptomatic Blastocystis infection. The high prevalence of Blastocystis infection within households suggested transmission between humans and their pets. Subtyping analysis of SSU rDNA alone in Blastocystis does not appear to predict pathogenicity.


Subject(s)
Blastocystis Infections/parasitology , Blastocystis/classification , Cat Diseases/parasitology , Dog Diseases/parasitology , Adult , Aged , Animals , Antiprotozoal Agents/therapeutic use , Asymptomatic Diseases , Biopsy , Blastocystis/isolation & purification , Blastocystis/pathogenicity , Blastocystis Infections/drug therapy , Blastocystis Infections/transmission , Blastocystis Infections/veterinary , Carrier State/drug therapy , Carrier State/parasitology , Cat Diseases/drug therapy , Cats , Disease Reservoirs , Dog Diseases/drug therapy , Dogs , Family Health , Female , Humans , Intestinal Mucosa/parasitology , Intestinal Mucosa/pathology , Male , Metronidazole/therapeutic use , Middle Aged , Prospective Studies , Ribotyping , Treatment Failure , Trimethoprim, Sulfamethoxazole Drug Combination/therapeutic use , Young Adult
11.
Aust Vet J ; 88(7): 260-2, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20579031

ABSTRACT

OBJECTIVE: The first national abattoir survey of Cysticercus bovis ('beef measles') in cattle was conducted in February 2008. METHODS: During the data collection period, 493,316 cattle were subjected to standard postmortem procedures, including incision of the masseter and heart muscles. On-site veterinarians were asked to record the location of any C. bovis cysts, as well as the National Livestock Identification System ear tag numbers of infected animals. Veterinarians were asked to submit samples for laboratory confirmation by histology and polymerase chain reaction testing. RESULTS: Of the 23 samples submitted, none was positive for C. bovis by either diagnostic method. CONCLUSIONS: Occasional, isolated diagnoses of beef measles are still made in most states of Australia, but since the last regional surveys were conducted 30 years ago, when the estimated prevalence was 50 to 200 per 100,000 cattle slaughtered, the parasite has become extremely rare.


Subject(s)
Abattoirs , Cattle Diseases/epidemiology , Cysticercosis/veterinary , Animals , Australia/epidemiology , Cattle , Cattle Diseases/parasitology , Cysticercosis/epidemiology , Cysticercus/isolation & purification , Female , Male , Muscle, Skeletal/parasitology , Prevalence
12.
Neuroscience ; 154(4): 1562-7, 2008 Jul 17.
Article in English | MEDLINE | ID: mdl-18550290

ABSTRACT

The existence of a sex difference in several chronic pain syndromes and the fluctuation of symptoms during the menstrual cycle strongly suggest sex hormones are involved in pain processing. The mechanisms underlying these changes are not well understood. Using the colorectal distention model in the rat, we previously reported a sex difference in the response to distention [Ji Y, Murphy AZ, Traub RJ (2006) Sex differences in morphine induced analgesia of visceral pain are supraspinally and peripherally mediated. Am J Physiol Regul Integr Comp Physiol 291:R307-R314] and that ovariectomy decreased the responses to distention while estrogen replacement reversed the decrease [Ji Y, Murphy AZ, Traub RJ (2003) Estrogen modulates the visceromotor reflex and responses of spinal dorsal horn neurons to colorectal stimulation in the rat. J Neurosci 23:3908-3915], suggesting estrogen increases visceral nociception. In the present study we tested the hypothesis that the visceromotor response to colorectal distention fluctuates with the estrous cycle. Three measurements (vaginal smears, uterine tube weight and plasma estrogen concentration) were used to determine the estrous phase. Comparison of the visceromotor response threshold and magnitude was made between proestrus and metestrus/diestrus. Our experiment demonstrated that the distention threshold was significantly lower in proestrus (median: 15 mm Hg) as compared with metestrus/diestrus (median: 25 mm Hg); and the magnitude of the visceromotor response to graded intensities of colorectal distentions (20, 40, 60, 80 mm Hg) was significantly higher in proestrus. The results indicate that the visceromotor response fluctuates with estrous phase, providing evidence for endogenous estrogen modulation of visceral nociceptive processing that could contribute to sex differences.


Subject(s)
Estrous Cycle/physiology , Pain Threshold/physiology , Pain/physiopathology , Animals , Area Under Curve , Colon/physiopathology , Dilatation, Pathologic , Electromyography , Estrogens/blood , Female , Rats , Rats, Sprague-Dawley , Rectum/physiopathology
13.
Exp Parasitol ; 116(2): 111-9, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17266951

ABSTRACT

Blastocystis is a prevalent single-celled enteric parasite of unresolved clinical significance. Efforts based on molecular methodologies to establish whether pathogenicity is linked to specific isolates of the genetically diverse genus of Blastocystis have been scarce and so far yielded ambiguous results which can be difficult to interpret. To alleviate some of the problems related to unravelling the molecular epidemiology of Blastocystis infections we developed and evaluated a simple and high-throughput sequence analysis (SQA) pyrosequencing technique based on the detection of genotype-specific nucleotide polymorphisms in the 18S small subunit rRNA gene for a rapid and cost-effective post-PCR screening of Blastocystis genotypes. The method was effectively capable of genotyping 48/48 isolates positive by nested PCR in approximately one hour, and in 94% of the cases the isolate detected by PCR and pyrosequencing was also detected by one of two different PCR assays with subsequent dideoxy sequencing.


Subject(s)
Blastocystis Infections/parasitology , Blastocystis/classification , DNA, Protozoan/chemistry , Sequence Analysis, DNA/methods , Animals , Base Sequence , Blastocystis/genetics , Cost-Benefit Analysis , Genotype , Humans , Molecular Sequence Data , Polymerase Chain Reaction , RNA, Protozoan/genetics , RNA, Ribosomal, 18S/genetics , Sequence Analysis, DNA/economics , Time Factors
14.
Parasitology ; 134(Pt 1): 113-9, 2007 Jan.
Article in English | MEDLINE | ID: mdl-16987431

ABSTRACT

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.


Subject(s)
Ancylostoma/classification , Ancylostomiasis/veterinary , Cat Diseases/parasitology , Dog Diseases/parasitology , Ancylostoma/anatomy & histology , Ancylostoma/genetics , Ancylostomiasis/parasitology , Animals , Australia , Cats , Cricetinae , DNA, Ribosomal Spacer/genetics , Dogs , Female , India , Male , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction
15.
Parasitology ; 134(Pt 3): 359-67, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17052374

ABSTRACT

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.


Subject(s)
Blastocystis Infections/transmission , Blastocystis/classification , Feces/parasitology , Polymerase Chain Reaction/methods , Zoonoses/transmission , Animals , Animals, Zoo/parasitology , Australia , Blastocystis/genetics , Blastocystis/isolation & purification , Blastocystis Infections/parasitology , Cat Diseases/parasitology , Cat Diseases/transmission , Cats , Culture Media , Dog Diseases/parasitology , Dog Diseases/transmission , Dogs , Humans , Marsupialia/parasitology , Primates/parasitology , Sequence Analysis, DNA , Thailand , Zoonoses/parasitology
16.
Radiat Prot Dosimetry ; 121(2): 202-7, 2006.
Article in English | MEDLINE | ID: mdl-17142822

ABSTRACT

Lung phantoms have been manufactured using commercially available, polyurethane foam products. Some of these materials are no longer available; therefore, a new lung tissue substitute was developed. The elemental composition and radiological properties of the new lung tissue substitute are described in this paper. Because the lung tissue substitute will be used to manufacture phantom lungs that will be used to evaluate chest counting systems, it is necessary to know the radiological properties of the material. These properties must be compared with reference materials and materials that have been used for lung phantoms in the past. The radiological properties of interest include the electron density, mean excitation energy, electron stopping power and photon mass attenuation coefficients. In all these properties, the calculated values for the new lung tissue substitute closely matched the calculated values of ICRU Publication 44 lung tissue. Good agreement was also found when the new lung tissue substitute was compared with the Griffith lung tissue substitute described by the ICRU. The new material was determined to be an excellent lung tissue substitute.


Subject(s)
Biomimetic Materials/chemistry , Lung/physiology , Photons , Electrons , Humans , Lung/radiation effects , Phantoms, Imaging
17.
Int J Parasitol ; 35(11-12): 1295-307, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16143334

ABSTRACT

Sound application of molecular epidemiological principles requires working knowledge of both molecular biological and epidemiological methods. Molecular tools have become an increasingly important part of studying the epidemiology of infectious agents. Molecular tools have allowed the aetiological agent within a population to be diagnosed with a greater degree of efficiency and accuracy than conventional diagnostic tools. They have increased the understanding of the pathogenicity, virulence, and host-parasite relationships of the aetiological agent, provided information on the genetic structure and taxonomy of the parasite and allowed the zoonotic potential of previously unidentified agents to be determined. This review describes the concept of epidemiology and proper study design, describes the array of currently available molecular biological tools and provides examples of studies that have integrated both disciplines to successfully unravel zoonotic relationships that would otherwise be impossible utilising conventional diagnostic tools. The current limitations of applying these tools, including cautions that need to be addressed during their application are also discussed.


Subject(s)
Molecular Epidemiology , Parasites/genetics , Parasitic Diseases/epidemiology , Animals , Disease Vectors , Epidemiologic Studies , Host-Parasite Interactions , Humans , Parasitic Diseases/diagnosis , Parasitic Diseases/transmission , Zoonoses
18.
Radiat Prot Dosimetry ; 110(1-4): 725-9, 2004.
Article in English | MEDLINE | ID: mdl-15353738

ABSTRACT

A project headed by Washington Group International is meant to design the Pit Disassembly and Conversion Facility (PDCF) to convert the plutonium pits from excessed nuclear weapons into plutonium oxide for ultimate disposition. Battelle staff are performing the shielding calculations that will determine appropriate shielding so that the facility workers will not exceed target exposure levels. The target exposure levels for workers in the facility are 5 mSv y(-1) for the whole body and 100 mSv y(-1) for the extremity, which presents a significant challenge to the designers of a facility that will process tons of radioactive material. The design effort depended on shielding calculations to determine appropriate thickness and composition for glove box walls, and concrete wall thicknesses for storage vaults. Pacific Northwest National Laboratory (PNNL) staff used ORIGEN-S and SOURCES to generate gamma and neutron source terms, and Monte Carlo (computer code for) neutron photon (transport) (MCNP-4C) to calculate the radiation transport in the facility. The shielding calculations were performed by a team of four scientists, so it was necessary to develop a consistent methodology. There was also a requirement for the study to be cost-effective, so efficient methods of evaluation were required. The calculations were subject to rigorous scrutiny by internal and external reviewers, so acceptability was a major feature of the methodology. Some of the issues addressed in the development of the methodology included selecting appropriate dose factors, developing a method for handling extremity doses, adopting an efficient method for evaluating effective dose equivalent in a non-uniform radiation field, modelling the reinforcing steel in concrete, and modularising the geometry descriptions for efficiency. The relative importance of the neutron dose equivalent compared with the gamma dose equivalent varied substantially depending on the specific shielding conditions and lessons were learned from this effect. This paper addresses these issues and the resulting methodology.


Subject(s)
Algorithms , Facility Design and Construction/methods , Neutrons , Occupational Exposure/analysis , Radiation Protection/methods , Radiometry/methods , Risk Assessment/methods , Body Burden , Calibration , Decontamination/methods , Environmental Monitoring/instrumentation , Environmental Monitoring/methods , Equipment Design , Equipment Failure Analysis/methods , Humans , Models, Biological , Quality Assurance, Health Care/methods , Radiation Dosage , Radiation Protection/instrumentation , Radiometry/instrumentation , Relative Biological Effectiveness , Reproducibility of Results , Risk Factors , Safety Management/methods , Sensitivity and Specificity , United States
19.
Parasitology ; 128(Pt 3): 253-62, 2004 Mar.
Article in English | MEDLINE | ID: mdl-15080083

ABSTRACT

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).


Subject(s)
Dog Diseases/parasitology , Giardia/growth & development , Giardiasis/parasitology , Giardiasis/veterinary , Zoonoses/parasitology , Adolescent , Adult , Animals , Base Sequence , Child , Child, Preschool , DNA, Protozoan/chemistry , DNA, Protozoan/genetics , Dogs , Female , Giardia/genetics , Giardiasis/epidemiology , Giardiasis/transmission , Humans , India/epidemiology , Infant , Male , Middle Aged , Molecular Sequence Data , Peptide Elongation Factor 1/chemistry , Peptide Elongation Factor 1/genetics , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal/chemistry , RNA, Ribosomal/genetics , Sequence Alignment , Sequence Analysis, DNA , Triose-Phosphate Isomerase/chemistry , Triose-Phosphate Isomerase/genetics , Zoonoses/epidemiology , Zoonoses/transmission
20.
Epidemiol Infect ; 132(1): 137-44, 2004 Jan.
Article in English | MEDLINE | ID: mdl-14979599

ABSTRACT

The prevalence of colonization with the anaerobic intestinal spirochaetes Brachyspira aalborgi and Brachyspira pilosicoli was investigated in humans (n = 316) and dogs (n = 101) living on three tea estates in Assam, India. Colonization was detected using PCR on DNA from faeces. Nineteen (6%) human faecal samples contained B. aalborgi DNA, 80 (25.3%) contained B. pilosicoli DNA, and 10 (3.2%) contained DNA from both species. One canine sample contained DNA from B. pilosicoli. Significant factors for B. aalborgi colonization in logistic regression were: infection of family members with B. aalborgi (P < 0.001), being a resident of Balipara (P = 0.03), and use of water treatment (P = 0.03). For B. pilosicoli, significant factors were: other family members being positive for B. pilosicoli (P < 0.001), water obtained from a well (P = 0.006), water treatment (P = 0.03), and not having visited a doctor in the previous 12 months (P = 0.03).


Subject(s)
Agricultural Workers' Diseases/microbiology , Agricultural Workers' Diseases/veterinary , Dog Diseases/microbiology , Spirochaetales Infections , Spirochaetales Infections/microbiology , Spirochaetales Infections/veterinary , Tea , Agricultural Workers' Diseases/epidemiology , Analysis of Variance , Animals , Brachyspira/classification , Brachyspira/genetics , Colony Count, Microbial , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Developing Countries , Dog Diseases/epidemiology , Dogs , Feces/microbiology , Humans , India/epidemiology , Logistic Models , Polymerase Chain Reaction , Population Surveillance , Prevalence , Residence Characteristics/statistics & numerical data , Risk Factors , Rural Health/statistics & numerical data , Serotyping , Spirochaetales Infections/epidemiology , Surveys and Questionnaires , Water Microbiology
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