scholarly journals Cross-genera amplification and identification of Colpodella sp. with Cryptosporidium primers in fecal samples of zoo felids from northeast China

2023 ◽  
Vol 83 ◽  
Author(s):  
S. Hussain ◽  
X. Li ◽  
S. M. Bukhari ◽  
M. Zhou ◽  
S. Ahmad ◽  
...  
Keyword(s):  
Northeast China ◽  
Diarrheal Disease ◽  
Clinical Epidemiology ◽  
Domestic Animals ◽  
Human Pathogens ◽  
Specific Primers ◽  
Accurate Identification ◽  
Fecal Samples ◽  
Identification Of Species ◽  
Water Borne

Abstract The protozoans include many intracellular human pathogens. Accurate detection of these pathogens is necessary to treat the diseases. In clinical epidemiology, molecular identification of protozoan is considered a more reliable and rapid method for identification than microscopy. Among these protozoans, Cryptosporidium considered being one of the important water-borne zoonotic pathogens and a major cause of a diarrheal disease named cryptosporidiosis in humans, domestic animals, and wild animals. This study was aimed to identify Cryptosporidium in zoo felids (N= 56) belonging to different zoo of China, but accidentlly Colpodella was encountered in the zoo felids sample and phylogenetic data confirmed this unexpected amplification from fecal samples using two-step nested-PCR. Phylogenetic analysis revealed the fact about the specific primers used previously by many researchers and cross-genera amplification. We came to know that genetically sequenced amplicon gives more accurate identification of species. This study suggests more investigation on Colpodella which has been neglected previously but gains the attention of researchers after identified from humans and animals and has been known to correlate with neurological symptoms in patients.

scholarly journals Prevalence and molecular detection of shiga toxin producing Escherichia coli from diarrheic cattle

2016 ◽  
Vol 14 (1) ◽  
pp. 63-68 ◽  
Author(s):  
MM Akter ◽  
S Majumder ◽  
KH MNH Nazir ◽  
M Rahman
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Molecular Detection ◽  
Chain Reaction ◽  
Specific Primers ◽  
Fecal Samples ◽  
E Coli ◽  
Uremic Syndrome ◽  
Polymerase Chain ◽  
Positive Isolate

Shiga toxin-producing Escherichia coli (STEC) are zoonotically important pathogen which causes hemorrhagic colitis, diarrhea, and hemolytic uremic syndrome in animals and humans. The present study was designed to isolate and identify the STEC from fecal samples of diarrheic cattle. A total of 35 diarrheic fecal samples were collected from Bangladesh Agricultural University (BAU) Veterinary Teaching Hospital. The samples were primarily examined for the detection of E. coli by cultural, morphological and biochemical characteristics, followed by confirmation of the isolates by Polymerase Chain Reaction (PCR) using gene specific primers. Later, the STEC were identified among the isolated E. coli through detection of Stx-1 and Stx-2 genes using duplex PCR. Out of 35 samples, 25 (71.43%) isolates were confirmed to be associated with E. coli, of which only 7 (28%) isolates were shiga toxin producers, and all of them were positive for Stx-1. However, no Stx-2 positive isolate could be detected. From this study, it may be concluded that cattle can act as a reservoir of STEC which may transmit to human or other animals.J. Bangladesh Agril. Univ. 14(1): 63-68, June 2016

scholarly journals Developing, Modifying, and Validating a TaqMan Real-Time PCR Technique for Accurate Identification of Leishmania Parasites Causing Most Leishmaniasis in Iran

2021 ◽  
Vol 11 ◽  
Author(s):  
Reza Fotouhi-Ardakani ◽  
Seyedeh Maryam Ghafari ◽  
Paul Donald Ready ◽  
Parviz Parvizi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Leishmania Major ◽  
Taqman Probe ◽  
Amino Acid Permease ◽  
Specific Primers ◽  
Accurate Identification ◽  
Parasitic Load ◽  
Species Specific

Many laboratory methods are used to diagnose leishmaniasis because it is characterized by varied symptoms and caused by different Leishmania species. A quantitative real-time PCR method based on a TaqMan probe was developed and modified for accurate identification of human cutaneous leishmaniasis (caused by Leishmania major or Leishmania tropica) from endemic areas of Iran. Two gene regions of amino acid permease 3 (AAP3) and cytochrome oxidase II (COII) were considered. Six new sets of species-specific primers and probes were designed. A total of 123 samples were examined and employed to evaluate and validate real-time PCR. According to parasitic load of the genesig®Leishmania Advanced Standard Kit, a serial dilution of purified plasmid (2–2×107 copies/reaction) was prepared under the same conditions for both genes. Specific primers and probes were able to detect three and six parasite copies in AAP3 and COII genes, respectively, and were able to detect three copies of parasites for L. major and L. tropica. The sensitivities of the reference kit and our method were 98.7 and 98.1%, respectively, and specificity was 100% for detecting parasite genomes in all assays. Designed primers and probes performed well in terms of efficiency and regression coefficient. For AAP3 and COII genes, respectively, the linear log range was 7 and the correlation coefficient (R2) was 0.749 and 0.996 for the reference kit using the standard generated curve and 0.98 and 0.96 with serial dilutions of parasite DNA. This research detected L. major and L. tropica definitely and opens the horizon for the other scientists in the multiplex reactions in designing and optimization of the conditions in silico and in vivo.

Proteomic fingerprinting for the fast and accurate identification of species in the Polyporoid and Hymenochaetoid fungi clades

2019 ◽  
Vol 203 ◽  
pp. 103390 ◽  
Author(s):  
Luiz Marcelo Ribeiro Tomé ◽  
Fernanda Badotti ◽  
Gabriella Borba Netto Assis ◽  
Paula Luize Camargos Fonseca ◽  
Genivaldo Alves da Silva ◽  
...  
Keyword(s):  
Accurate Identification ◽  
Identification Of Species ◽  
Proteomic Fingerprinting

scholarly journals Rotavirus C: prevalence in suckling piglets and development of virus-like particles to assess the influence of maternal immunity on the disease development

2019 ◽  
Vol 50 (1) ◽  
Author(s):  
Juliet Chepngeno ◽  
Annika Diaz ◽  
Francine C. Paim ◽  
Linda J. Saif ◽  
Anastasia N. Vlasova
Keyword(s):  
Cell Culture ◽  
Diarrheal Disease ◽  
Economic Losses ◽  
Disease Development ◽  
Fecal Samples ◽  
Virus Like Particles ◽  
Maternal Immunity ◽  
The Us ◽  
Suckling Piglets ◽  
Major Impediment

Abstract Rotavirus C (RVC) has been detected increasingly in humans and swine in different countries, including the US. It is associated with significant economic losses due to diarrheal disease in nursing piglets. In this study we aimed: (1) to determine the prevalence of RVC in healthy and diarrheic suckling piglets on US farms; and (2) to evaluate if maternal antibody (Ab) levels were associated with protection of newborn suckling piglets against RVC. There was a significantly higher prevalence (p = 0.0002) of litters with diarrhea born to gilts compared with those born to multiparous sows. Of 113 nursing piglet fecal samples tested, 76.1% were RVC RNA positive. Fecal RVC RNA was detected in significantly (p = 0.0419) higher quantities and more frequently in piglets with diarrhea compared with healthy ones (82.5 vs. 69.9%). With the exception of the historic strain Cowden (G1 genotype), field RVC strains do not replicate in cell culture, which is a major impediment for studying RVC pathogenesis and immunity. To circumvent this, we generated RVC virus-like particles (VLPs) for Cowden (G1), RV0104 (G3) and RV0143 (G6) and used them as antigens in ELISA to detect swine RVC Abs in serum and milk from the sows. Using RVC-VLP Ab ELISA we demonstrated that sows with diarrheic litters had significantly lower RVC IgA and IgG Ab titers in milk compared to those with healthy litters. Thus, our data suggest that insufficient lactogenic protection provided by gilts plays a key role in the development of and the increased prevalence of clinical RVC disease.

scholarly journals Prevalence, Environmental Loading, and Molecular Characterization of Cryptosporidium and Giardia Isolates from Domestic and Wild Animals along the Central California Coast

2012 ◽  
Vol 78 (24) ◽  
pp. 8762-8772 ◽  
Author(s):  
Stori C. Oates ◽  
Melissa A. Miller ◽  
Dane Hardin ◽  
Patricia A. Conrad ◽  
Ann Melli ◽  
...  
Keyword(s):  
Beef Cattle ◽  
Parasite Load ◽  
Domestic Animals ◽  
Host Group ◽  
California Coast ◽  
Fecal Samples ◽  
Content Type ◽  
Central California ◽  
Animal Host ◽  
Wild Canids

ABSTRACTThe risk of disease transmission from waterborne protozoa is often dependent on the origin (e.g., domestic animals versus wildlife), overall parasite load in contaminated waterways, and parasite genotype, with infections being linked to runoff or direct deposition of domestic animal and wildlife feces. Fecal samples collected from domestic animals and wildlife along the central California coast were screened to (i) compare the prevalence and associated risk factors for fecal shedding ofCryptosporidiumandGiardiaspecies parasites, (ii) evaluate the relative importance of animal host groups that contribute to pathogen loading in coastal ecosystems, and (iii) characterize zoonotic and host-specific genotypes. Overall, 6% of fecal samples tested during 2007 to 2010 were positive forCryptosporidiumoocysts and 15% were positive forGiardiacysts. Animal host group and age class were significantly associated with detection ofCryptosporidiumandGiardiaparasites in animal feces. Fecal loading analysis revealed that infected beef cattle potentially contribute the greatest parasite load relative to other host groups, followed by wild canids. Beef cattle, however, shed host-specific, minimally zoonoticCryptosporidiumandGiardia duodenalisgenotypes, whereas wild canids shed potentially zoonotic genotypes, includingG. duodenalisassemblages A and B. Given that the parasite genotypes detected in cattle were not zoonotic, the public health risk posed by protozoan parasite shedding in cattle feces may be lower than that posed by other animals, such as wild canids, that routinely shed zoonotic genotypes.

Tree kangaroo molecular systematics based on partial cytochrome b sequences: are Matschie's tree kangaroo (Dendrolagus matschiei) and Goodfellow's tree kangaroo (D. goodfellowi buergersi) sister taxa?

2012 ◽  
Vol 34 (1) ◽  
pp. 18 ◽  
Author(s):  
Thomas J. McGreevy ◽  
Lisa Dabek ◽  
Thomas P. Husband
Keyword(s):  
Mitochondrial Dna ◽  
Papua New Guinea ◽  
Cytochrome B ◽  
Molecular Systematics ◽  
Phylogenetic Analyses ◽  
New Guinea ◽  
Critically Endangered ◽  
Evolutionary Relationships ◽  
Accurate Identification ◽  
Identification Of Species

New Guinea tree kangaroos (Dendrolagus spp.) are unique arboreal macropodid marsupials mainly listed as critically endangered or endangered. The molecular systematics of Dendrolagus has not been fully resolved and is critical for the accurate identification of species and their evolutionary relationships. Matschie’s tree kangaroo (D. matschiei) and Goodfellow’s tree kangaroo (D. goodfellowi buergersi) share numerous morphological, physiological, and behavioural traits. We analysed the partial mitochondrial DNA cytochrome b gene for D. matschiei (n = 67), D. g. buergersi (n = 8), D. goodfellowi unidentified ssp. (n = 8), golden-mantled tree kangaroo (D. g. pulcherrimus; n = 1), and two additional New Guinea Dendrolagus taxa to determine whether D. matschiei and D. g. buergersi are sister taxa. D. matschiei and D. g. buergersi were not placed as sister taxa in our phylogenetic analyses; however, we were unable to analyse a known sample from a D. g. goodfellowi. We found initial genetic evidence that D. matschiei and the Lowland tree kangaroo (D. spadix) are sister taxa – they may have diverged after the formation of the Huon Peninsula of Papua New Guinea. Our results also support the elevation of D. g. pulcherrimus to a full species. An improved understanding of Dendrolagus molecular systematics will contribute substantially to their conservation.

Analysis of genetic similarities among species of Fragaria, Potentilla, and Duchesnea found in northwest Argentina by using morphological, anatomical, and molecular characters

2000 ◽  
Vol 78 (4) ◽  
pp. 547-556 ◽  
Author(s):  
Marta Ontivero ◽  
Marta Arias ◽  
Juan Diaz Ricci ◽  
Judith Babot ◽  
Patricia Albornoz ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Molecular Techniques ◽  
Dna Polymorphisms ◽  
Randomly Amplified Polymorphic Dna ◽  
Specific Primers ◽  
Northwest Argentina ◽  
Identification Of Species ◽  
Wild Strawberry ◽  
Anatomical Characters ◽  
Molecular Characters

Morphological, anatomical, and molecular techniques were used to characterize wild strawberry and wild strawberry-like species in northwest Argentina. Characteristics of leaves, flowers, runners, achenes, and genomic DNA polymorphisms were used to analyze similarities among Potentilla tucumanensis Castagnaro & Arias, Duchesnea indica (Andr.) Focke, and Fragaria vesca L. Comparison of phenograms obtained by using morphological and anatomical traits or genomic DNA characters revealed similar clustering of the species. Both phenograms suggest that D. indica is more closely related to P. tucumanensis than to F. vesca. Using the randomly amplified polymorphic DNA (RAPD) technique with specific primers, we detected polymorphic bands that permit the identification of P. tucumanensis, D. indica, and F. vesca. In addition, we report new morphological and anatomical characters that can be used as diagnostic traits for better identification of species in reproductive and vegetative states.Key words: Fragaria, Potentilla, Duchesnea, RAPD, DNA fingerprinting, morphological traits, anatomical traits.

scholarly journals A Pitfall in Mouse Norovirus (MNV) Detection in Fecal Samples Using RT-PCR, and Construction of New MNV-Specific Primers

2013 ◽  
Vol 62 (2) ◽  
pp. 127-135 ◽  
Author(s):  
Masaru TAJIMA ◽  
Yuko KOTANI ◽  
Tsutomu KUROSAWA ◽  
Masayuki MIYASAKA
Keyword(s):  
Rt Pcr ◽  
Specific Primers ◽  
Fecal Samples

Shedding new light on old species identifications: morphological and genetic evidence suggest a need for conservation status review of the critically endangered bat, Saccolaimus saccolaimus

2009 ◽  
Vol 36 (6) ◽  
pp. 496 ◽  
Author(s):  
Damian J. Milne ◽  
Felicity C. Jackling ◽  
Manpreet Sidhu ◽  
Belinda R. Appleton
Keyword(s):  
Threatened Species ◽  
Conservation Status ◽  
Critically Endangered ◽  
Similar Species ◽  
Museum Specimens ◽  
Museum Collections ◽  
Species Classification ◽  
Accurate Identification ◽  
Identification Of Species ◽  
Vital Component

Information based on the accurate identification of species is a vital component for achieving successful outcomes of biodiversity conservation and management. It is difficult to manage species that are poorly known or that are misidentified with other similar species. This is particularly problematic for rare and threatened species. Species that are listed under endangered species classification schemes need to be identified accurately and categorised correctly so that conservation efforts are appropriately allocated. In Australia, the emballonurid Saccolaimus saccolaimus is currently listed as ‘Critically Endangered’. On the basis of new observations and existing museum specimens, we used a combination of genetic (mitochondrial DNA sequence) and morphological (pelage characteristics, dig III : phalanx I length ratio, inter-upper canine distance) analyses to identify six new geographic records for S. saccolaimus, comprising ~100 individuals. Our analyses also suggested that there are likely to be more records in museum collections misidentified as S. flaviventris specimens. The external morphological similarities to S. flaviventris were addressed and genetic, morphological and echolocation analyses were used in an attempt to provide diagnostic characters that can be used to readily identify the two species in the field. We recommend genetic testing of all museum specimens of Australian Saccolaimus to clarify species’ distributions and provide data for reassessing the conservation status for both S. saccolaimus and S. flaviventris. Museum curators, taxonomists and wildlife managers need to be aware of potential species misidentifications, both in the field and laboratory. Misidentifications that result in misclassification of both threatened and non-threatened species can have significant implications.

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