Accepted ManuscriptThis is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. faecal samples. An internal amplification control (IAC) was also developed and included in this assay. The qPCR assay was compared with an 18S nested PCR assay for sensitivity and specificity. The analytical sensitivity for the qPCR assay was 1 oocyst and 1-10 oocysts for the 18S assay. Evaluation of analytical specificity of the qPCR assay revealed no crossreactions with other genera and detected all C. parvum and C. hominis isolates correctly. The diagnostic sensitivity and specificity of the qPCR was 100% compared to 96.9% and 98.4% Specific and quantitative detection and identification ofrespectively for the 18S assay. The qPCR assay was also highly reproducible with RSD (relative standard deviation) values of 1.4-9.4%, when the assay was performed by four different technicians. When tested on water samples, the qPCR assay was more sensitive than the 18S assay, detecting positives in 37 of 138 water samples compared to 35 for the 18Slocus. This qPCR assay should be a valuable tool for the detection and differentiation of C.hominis and C. parvum in both clinical and environmental samples.
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Page 1 of 28A c c e p t e d M a n u s c r i p t The prevalence of Cryptosporidium in sheep in the eastern states of Australia has not been 21 well described, therefore a study of the prevalence, oocyst concentration, species and subtypes of 22Cryptosporidium were assessed from lamb faecal samples at three sampling periods (weaning, post-23 weaning and pre-slaughter) from eight farms across South Australia, New South Wales, Victoria 24 and Western Australia. A total of 3,412 faecal samples were collected from approximately 1,182 25 lambs across the 4 states and screened for the presence of Cryptosporidium using a quantitative 26 PCR (qPCR) at the actin locus. Positives were typed at the 18S locus and at a second locus using C. 27 parvum and C. hominis specific qPCR primers. The overall prevalence was 16.9% (95% CI: 15.6-28 18.1%) and of the 576 positives, 500 were successfully genotyped. In general, the prevalence of 29Cryptosporidium was higher in WA than the eastern states. Cryptosporidium prevalence peaked at 30 43.9% and 37.1% at Pingelly (WA2) and Arthur River (WA1) respectively during weaning and at 31
Please cite this article as: Koinari, M., Karl, S., Ng-Hublin, J., Lymbery, A.J., Ryan, U.M., Identification of novel and zoonotic Cryptosporidium species in fish from Papua New Guinea, Veterinary Parasitology (2013), http://dx.doi.org/10. 1016/j.vetpar.2013.08.031 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. maracellus and oblong silver biddy, Gerres oblongus), giving an overall prevalence of 1.14 % 26 (95 % CI: 0.3 % -2 %, n=7/614). Of the seven positive isolates, five were identified as C. 27 parvum and two were a novel piscine genotype, which we have named piscine genotype 8. 28Piscine genotype 8 was identified in two marine oblong silver biddies and exhibited 4.3 % 29 genetic distance from piscine genotype 3 at the 18S locus. Further subtyping of C. parvum 30 isolates at the 60 kDa glycoprotein (gp60) locus identified 3 C. parvum subtypes (IIaA14G2R1, 31 IIaA15G2R1 and IIaA19G4R1) all of which are zoonotic and a C. hominis subtype (IdA15G1). 32The zoonotic Cryptosporidium were identified in fish samples from all three groups; cultured 33 and wild freshwater and wild marine fish. Detection of Cryptosporidium among aquaculture 34 fingerlings warrants further research to gain a better understanding of the epidemiology of 35Cryptosporidium infection in cultured fish. The identification of zoonotic Cryptosporidium 36 genotypes in fish from PNG has important public health implications and should be investigated 37 further. 38
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.
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