Characterization of Corynebacterium diphtheriae isolates from infected skin lesions in the Northern Territory of Australia

Corynebacterium diphtheriae is commonly isolated from cutaneous skin lesions in the Northern Territory of Australia. We prospectively assessed 32 recent isolates from infected skin lesions, in addition to reviewing 192 isolates collected over 5 years for toxin status. No isolates carried the toxin gene. Toxigenic C. diphtheriae is now a rare occurrence in the Northern Territory.     

Field evaluation of repellent formulations containing deet and picaridin against mosquitoes in Northern Territory, Australia

Field efficacy of repellent formulations containing picaridin (1-methyl-propyl 2-(2-hydroxyethyl)-1-piperidinecarboxylate) or deet (N,N,-diethyl-3-methylbenzamide) against mosquitoes in Northern Territory, Australia, was evaluated. The following repellent treatments were evaluated: 19.2% picaridin (Autan Repel Army 20), a solution of 20% deet in ethanol, and 35% deet in a gel (Australian Defense Force [ADF]). The predominant mosquito species were Culex annulirostris Skuse (57.8%), Anopheles merankensis Venhuis (15.4%), and Anopheles bancroftii Giles (13.2%). The protection provided by repellents against Anopheles spp. was relatively poor, with 19.2% picaridin and ADF deet providing >95% protection for only 1 h, whereas 20% deet provided <95% protection at 1 h after repellent application. In contrast, the repellents provided good protection against Cx. annulirostris, with 19.2% picaridin providing >95% protection for 5 h and both deet formulations providing >95% protection for 7 h when collections ceased. This study provides additional field data showing tolerance of Anopheles spp. for repellents. The response of field populations of Cx. annulirostris, an important vector of arboviruses in Australia, to repellents containing deet and picaridin is reported for the first time.     

First surveillance and molecular identification of the Cryptosporidium skunk genotype and Cryptosporidium parvum in wild raccoons (Procyon lotor) in Osaka,Japan

Parasitology Research - Recent research suggests that raccoons (Procyon lotor) can transmit several important pathogens affecting humans, including protozoans. In Japan, the number of wild raccoons...     

Arbovirus models to provide practical management tools for mosquito control and disease prevention in the Northern Territory, Australia

Ross River virus (RRV) causes the most common human arbovirus disease in Australia. Although the disease is nonfatal, the associated arthritis and postinfection fatigue can be debilitating for many months, impacting on workforce participation. We sought to create an early-warning system to notify of approaching RRV disease outbreak conditions for major townships in the Northern Territory. By applying a logistic regression model to meteorologic factors, including rainfall, a postestimation analysis of sensitivity and specificity can create rainfall cut-points. These rainfall cut-points indicate the rainfall level above which previous epidemic conditions have occurred. Furthermore, rainfall cut-points indirectly adjust for vertebrate host data from the agile wallaby (Macropus agilis) as the life cycle of the agile wallaby is intricately meshed with the wet season. Once generated, cut-points can thus be used prospectively to allow timely implementation of larval survey and control measures and public health warnings to preemptively reduce RRV disease incidence. Cut-points are location specific and have the capacity to replace previously used models, which require data management and input, and rarely provide timely notification for vector control requirements and public health warnings. These methods can be adapted for use elsewhere.     

Human Cryptosporidiosis Diagnosed in Western Australia: a Mixed Infection with Cryptosporidium meleagridis,the Cryptosporidium Mink Genotype,and an Unknown Cryptosporidium Species

This report describes a case of cryptosporidiosis from an immunocompetent patient from Perth, Western Australia, suffering from diarrhea and a spectrum of other symptoms. Molecular identification revealed that this patient was infected with three Cryptosporidium species—Cryptosporidium meleagridis, the Cryptosporidium mink genotype, and an unknown Cryptosporidium species.     

An outbreak of trichinellosis in farmed wild boar in Finland

Nine farmed wild boar out of 25 slaughtered from a single farm were condemned at meat inspection because of trichinellosis. With RAPD-PCR, Trichinella spiralis was identified in all positive wild boar. Out of the available serum samples (n=7), all wild boar which had failed the meat inspection showed seroconversion in ELISA and Western blotting, as did one additional animal which had passed the inspection. The animals became infected during an invasion of rats from an improperly closed dump near the farm. Unfortunately, by the time trichinellosis was discovered in the wild boar, the invasion had already been brought under control; thus, no samples from rats were available. However, having lived through the rat invasion was shown to be a risk factor for trichinellosis in wild boar (relative risk, RR=6.3). In wildlife samples from surrounding areas, sylvatic trichinellosis was found to be very common (74%; n= 19 red foxes). Intriguingly, the prevalent species in trichinella-positive foxes differed from that in wild boar, Trichinella nativa and T. spiralis being found in 12 foxes and in one fox, respectively.     

Unusual cryptosporidium species recovered from human faeces: first description of Cryptosporidium felis and Cryptosporidium 'dog type' from patients in England

DNA was extracted from faecal samples collected from 1680 patients in which Cryptosporidium oocysts were recognised by light microscopy. DNA from faeces from five of these patients failed to amplify by PCR three gene fragments--the Cryptosporidium oocyst wall protein (COWP) gene, the thrombospondin-related adhesive protein of Cryptosporidium-1 (TRAP-C1) gene and the thrombospondin-related adhesive protein of Cryptosporidium-2 (TRAP-C2) gene--with primers designed from C. parvum sequences. However, DNA from these five patients did amplify cryptosporidial 18S rDNA gene fragments and a heat-shock protein (HSP70) gene fragment was also amplified from four of them. The purpose of this study was to characterise further the Cryptosporidium associated with infection in these patients. DNA sequence analysis of 18S rDNA genes showed that four of these patients were infected by C. felis, and the remaining one by an as yet un-named Cryptosporidium species designated the 'dog type' (C. dt). Infection by C. felis was further confirmed in all four patients by DNA sequence analysis of the HSP70 gene. Oocysts present in all five samples reacted strongly with two anti-cryptosporidial oocyst monoclonal antibodies, except for the C. dt, which was tested with only one of the antibodies. Two of the patients infected by C. felis had underlying illness; one 8-year-old male had an undefined severe inherited underlying condition, and the second patient, a 32-year-old male, was HIV positive. Two of the remaining three patients (two females aged 1 and 2 years, respectively) were apparently immunocompetent (one infected with C. felis and one with the C. dt). No information was obtained for the fifth patient. The patient infected by C. dt had a recent history of travel to Africa. This is the first report of infection with these two Cryptosporidium species in immunocompetent patients, and in any patient in the UK.     

Molecular detection and genetic characterizations of Cryptosporidium spp. in farmed foxes,minks, and raccoon dogs in northeastern China

Parasitology Research - Cryptosporidium spp. are common intestinal protozoa causing diarrhea in humans and a variety of animal species. With the recent development of fur industry, a large number...     

Concurrent Cryptosporidium and Candida infections in a buffalo calf (Bubalus bubalis)

Cryptosporidiosis and candidiasis are opportunistic infections of neonates. An investigation was contemplated to study the enteric lesions associated with concurrent infections by the pathogens in a male bubaline calf, aged 8 weeks, which died on March 23, 2010. The gross lesions were comprised of hyperplastic changes and erosion of the microvillus surface of the enteric epithelium. The histological sections revealed invasion of the mucosal cells by two types (Cryptosporidium spp. and Candida spp.) of pathogens. The deeper layers of the intestine did not reveal appreciable architectural changes, except for the presence of inflammatory cellular elements in the lamina propria. The sections stained with Ziehl–Neelsen revealed bright red, oval to spherical bodies (3–5 μm), and those stained with Gomori's Methanamine silver nitrate revealed black-colored fungal spores and pseudohyphae. Based on morphological and in vitro cultural characteristics, the pathogens were identified as Cryptosporidium spp. and Candida spp., respectively. The lesions seem to be consequential events to synergistic invasion of the enterocytes by the pathogens. Complex changes in the microenvironment of the gut, incidental to in situ release of hydrolytic enzymes, toxins, and products of host–parasite interaction liberated at the site of infection by the pathogens, have been reported and discussed.     

Analysis of oocyst wall and sporozoite antigens from three Cryptosporidium species.

A comparison was made of the antigenic composition of oocyst walls and sporozoites from Cryptosporidium baileyi from turkeys, C. muris from rodents, and C. parvum from ruminants, employing immunoblotting and immunofluorescence. In immunoblotting, oocyst antigens were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting (immunoblotting) and detected with rabbit polyclonal anti-C. muris or -C. parvum antibodies or murine monoclonal antibodies developed against C. parvum. Immunofluorescence was used to investigate the reactivity of these monoclonal antibodies with air-dried excystation mixtures of sporozoites and oocysts of the different species. The results from both types of experiment indicated that the three Cryptosporidium species could be differentiated immunologically. In comparison, few antigenic differences were found between a number of isolates of C. parvum in immunoblotting. There was also evidence to suggest that C. parvum and C. baileyi were more closely related antigenically to one another than to C. muris.     

First report of a ranavirus associated with morbidity and mortality in farmed Chinese giant salamanders (Andrias davidianus)

From February to May 2010, an outbreak of disease occurred amongst farmed Chinese giant salamanders (Andrias davidianus) in Hanzhong County, Shanxi Province, China. Clinical signs included anorexia, lethargy, ecchymoses and swollen areas on the head and limbs, and skin ulceration. The aim of this study was to determine the cause of this disease. Necropsy examination revealed subcutaneous and intramuscular oedema, swollen and pale livers with multifocal haemorrhage, swollen kidneys with multifocal haemorrhage and distended fluid-filled intestines with areas of haemorrhage. Light microscopy revealed intracytoplasmic inclusions suggestive of a viral infection in a variety of organs, as well as degeneration and necrosis of these organs. Electron microscopy of ultrathin sections of the same tissues revealed iridovirus-like particles within the inclusions. Of the six specimens tested, all were positive for ranavirus major capsid protein (MCP) gene. Sequence alignments of the ranavirus MCP gene from these specimens showed 95-98% similarity with published ranavirus data. The virus, provisionally designated as Chinese giant salamander virus (CGSV), was isolated from tissue homogenates of diseased salamanders following inoculation of epithelioma papilloma cyprini cells. Sequence analysis of the MCP genes showed that the isolated virus was a ranavirus with marked sequence identity to other members of the genus Ranavirus. Koch's postulates were fulfilled by infecting healthy Chinese giant salamanders with the CGSV. These salamanders all died within 6-8 days. This is the first report of ranavirus infection associated with mass mortality in Chinese giant salamanders.     

Molecular characterization of Cryptosporidium species and genotypes in Chile

Knowledge of species and genotypes on Cryptosporidium involved in human transmission is fundamental in studying the epidemiology of cryptosporidiosis. This is the first study in molecular epidemiology of cryptosporidiosis that has been made in Chile. Cryptosporidium parvum species and genotypes involved in human and bovine cryptosporidiosis were determined by PCR-RFLP. Low prevalence of Cryptosporidium was found in humans and cattle. Of the 127 cattle studied, only in one was C. parvum detected . Of the 68 human fecal samples studied, four contained Cryptosporidium: two patients presented Cryptosporidium hominis and two C. parvum. Our results confirm that anthroponotic transmission is as common as zoonotic in Chile. These data can enable the establishment of the corresponding prevention and treatment measures.     

Phylogenic analysis of the M genes of influenza viruses isolated from free-flying water birds from their Northern Territory to Hokkaido, Japan

During 2000-2007, 218 influenza viruses of 28 different combinations of HA (H1-H13) and NA (N1-N9) subtypes were isolated from fecal samples of free-flying water birds at two distant lakes in Hokkaido, Japan. Phylogenic analysis of the matrix (M) genes of 67 strains, selected on the basis of their subtype combinations, revealed that A/duck/Hokkaido/W95/2006 (H10N8) was a reassortant whose M gene belonged to North American non-gull-avian and the other seven genes to Eurasian non-gull-avian lineages. The M genes of other 65 strains belonged to Eurasian non-gull-avian and the one to Eurasian-gull lineages. The M genes of 65 strains were grouped into three different sublineages, indicating that influenza viruses circulating in different populations of free-flying water birds have evolved independently in nature.     

Distribution of Cryptosporidium subtypes in humans and domestic and wild ruminants in Portugal

To investigate the transmission of cryptosporidiosis in Portugal, Cryptosporidium hominis and Cryptosporidium parvum from HIV-infected patients, cattle, and wild ruminants were characterized by sequence analysis of the 60-kDa glycoprotein (GP60) gene. Fourteen subtypes within nine subtype families were identified, and three of the subtype families (If, IIb, and IId) were restricted or largely limited to Portugal. Parasites from cattle from various regions in Portugal and wild ruminants in Lisbon showed limited genetic heterogeneity (only two subtype families). All wild ruminants had the same subtype, which was also the predominant subtype in cattle all over Portugal and was found in nine HIV-infected patients in Lisbon. Two other C. parvum subtypes were only restricted to limited locations. In contrast, human parasites displayed 13 subtypes in nine subtype families, with most of the infections caused by parasites in Ib, IIa, IIc, and IId families. Two of the C. parvum subtype families (IIc and IIb) had only been found in humans. The high overall parasite diversity and high percentage of C. hominis infections attributable to Ib and C. parvum infections to IId represent unique characteristics of Cryptosporidium transmission in humans in Portugal.