scholarly journals Rapid Detection of Equine Piroplasms Using Multiplex PCR and First Genetic Characterization of Theileria haneyi in Egypt

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1414
Author(s):  
Bassma S. M. Elsawy ◽  
Ahmed M. Nassar ◽  
Heba F. Alzan ◽  
Raksha V. Bhoora ◽  
Sezayi Ozubek ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Genetic Characterization ◽  
Simultaneous Detection ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Theileria Equi ◽  
Babesia Caballi ◽  
Pcr Targeting ◽  
First Time

Equine Piroplasmosis (EP) is an infectious disease caused by the hemoprotozoan parasites Theileria equi, Babesia caballi, and the recently identified species T. haneyi. Hereby, we used a multiplex PCR (mPCR) targeting the 18S rRNA gene of T. equi and B. caballi for the simultaneous detection of EP in Egyptian equids and examined the presence of T. haneyi infections in Egypt. Blood samples from 155 equids (79 horses and 76 donkeys) collected from different governorates of Egypt were examined by mPCR and PCR targeting T. hayeni. The mPCR method revealed a prevalence of T. equi of 20.3% in horses and of 13.1% in donkeys and a prevalence of B. caballi of 1.2% in horses. B. caballi was not detected in donkeys in the current study. The mPCR method also detected coinfections with both species (2.5% and 1.3% in horses and donkeys, respectively). Additionally, we report the presence of T. haneyi in Egypt for the first time in 53.1% of the horse and 38.1% of the donkey tested samples. Coinfection with T. haneyi and T. equi was found in 13.5% of the samples, while infection with the three EP species was found in 1.9% of the samples.

scholarly journals Genetic characterization of Strongyloides spp. from captive, semi-captive and wild Bornean orangutans (Pongo pygmaeus) in Central and East Kalimantan, Borneo, Indonesia

Parasitology ◽  
2011 ◽  
Vol 138 (11) ◽  
pp. 1417-1422 ◽  
Author(s):  
E. M. LABES ◽  
N. WIJAYANTI ◽  
P. DEPLAZES ◽  
A. MATHIS
Keyword(s):  
18S Rrna ◽  
Genetic Characterization ◽  
18S Rrna Gene ◽  
Rdna Locus ◽  
Rrna Gene ◽  
Fecal Material ◽  
East Kalimantan ◽  
The One ◽  
Bornean Orangutans

SUMMARYOrangutans (Pongo spp.), Asia's only great apes, are threatened in their survival due to habitat loss, hunting and infections. Nematodes of the genus Strongyloides may represent a severe cause of death in wild and captive individuals. In order to better understand which Strongyloides species/subspecies infect orangutans under different conditions, larvae were isolated from fecal material collected in Indonesia from 9 captive, 2 semi-captive and 9 wild individuals, 18 captive groups of Bornean orangutans and from 1 human working with wild orangutans. Genotyping was done at the genomic rDNA locus (part of the 18S rRNA gene and internal transcribed spacer 1, ITS1) by sequencing amplicons. Thirty isolates, including the one from the human, could be identified as S. fuelleborni fuelleborni with 18S rRNA gene identities of 98·5–100%, with a corresponding published sequence. The ITS1 sequences could be determined for 17 of these isolates revealing a huge variability and 2 main clusters without obvious pattern with regard to attributes of the hosts. The ITS1 amplicons of 2 isolates were cloned and sequenced, revealing considerable variability indicative of mixed infections. One isolate from a captive individual was identified as S. stercoralis (18S rRNA) and showed 99% identity (ITS1) with S. stercoralis sequences from geographically distinct locations and host species. The findings are significant with regard to the zoonotic nature of these parasites and might contribute to the conservation of remaining orangutan populations.

scholarly journals Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa

2009 ◽  
Vol 159 (2) ◽  
pp. 112-120 ◽  
Author(s):  
Raksha Bhoora ◽  
Linda Franssen ◽  
Marinda C. Oosthuizen ◽  
Alan J. Guthrie ◽  
Erich Zweygarth ◽  
...  
Keyword(s):  
South Africa ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Theileria Equi ◽  
Babesia Caballi ◽  
Sequence Heterogeneity

scholarly journals Surveillance of Giardia and Cryptosporidium in sewage from an urban area in Brazil

2019 ◽  
Vol 28 (2) ◽  
pp. 291-297 ◽  
Author(s):  
Felippe Danyel Cardoso Martins ◽  
Winni Alves Ladeia ◽  
Roberta dos Santos Toledo ◽  
João Luis Garcia ◽  
Italmar Teodorico Navarro ◽  
...  
Keyword(s):  
18S Rrna ◽  
Fragment Length Polymorphism ◽  
Sewage Water ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Protozoan Parasites ◽  
Treated Sewage ◽  
Multiple Species ◽  
Pcr Targeting

Abstract Cryptosporidium and Giardia are protozoan parasites that cause diarrhea in humans and animals. Molecular characterization of these pathogens in sewage may provide insight on their occurrence and prevalence in Brazil. This study aimed to investigate the presence of Giardia and Cryptosporidium in raw and treated sewage from Londrina, Paraná, Brazil. Samples were collected every two weeks during a year. Samples were concentrated, then DNA was extracted and subjected to a nested PCR targeting the Giardia 18S rRNA gene and the Cryptosporidium 18S rRNA gene. Species of Cryptosporidium were characterized by restriction fragment length polymorphism (RFLP). All raw sewage and 76% of the treated sewage were positive for Giardia; 84% of raw sewage samples and 8% of treated sewage were positive for Cryptosporidium. C. muris, C. hominis, C. baileyi, C. parvum and C. suis were detected in 100%, 19%, 9%, 9% and 4% of raw sewage, respectively. C. muris was the only species found in treated sewage. Multiple species of Cryptosporidium were present in 19.04% of the raw sewage. Treated sewage water can pose a threat to human health. The speciation of Cryptosporidium revealed the presence of non-common zoonotic species as C. suis and C. muris.

scholarly journals Molecular Investigation and Genotyping of Theileria equi and Babesia caballi in Horses in Mus Province, Turkey

2021 ◽  
Vol 71 (4) ◽  
pp. 2531
Author(s):  
B. OGUZ ◽  
N. ÖZDAL ◽  
M.S. DEGER ◽  
K. BICEK
Keyword(s):  
Phylogenetic Analyses ◽  
Epidemiological Data ◽  
The United States ◽  
18S Rrna Gene ◽  
Diagnostic Tools ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Theileria Equi ◽  
Babesia Caballi ◽  
Equine Piroplasmosis

Equine Piroplasmosis (EP) is a tick-borne disease caused by Theileria equi and Babesia caballi of the phylum Apicomplexa. In this study, 102 blood samples were randomly collected from the horses in Mus province of Turkey. PCR analysis, gene sequences, and phylogenetic analyses were carried out for detecting the presence and genotypic characteristics of species that cause piroplasmosis. Four (3.9%) of the 102 horses that were examined were found to be positive for T. equi, while B. caballi was not detected. Theileria equi isolates that were detected in the sequence analyses were found to be 100% identical to the isolates that were isolated from the horses in Turkey, the United States, and South Africa as well. In the phylogenetic analysis, all of the isolates were found to cluster with T. equi sequences in the genotype A. This study, in which we revealed intraspecies sequence heterogeneity of the parasite using the 18S rRNA gene region, provides important epidemiological data for equine piroplasmosis. However, we think that determining the characterization of genotypes that are common in different parts of our country is extremely important in terms of developing new diagnostic tools and vaccines.

scholarly journals First study of Cryptosporidium spp. occurrence in eared doves (Zenaida auriculata)

2019 ◽  
Vol 28 (3) ◽  
pp. 489-492
Author(s):  
Mércia de Seixas ◽  
Alessandra Taroda ◽  
Sérgio Tosi Cardim ◽  
João Pedro Sasse ◽  
Thais Agostinho Martins ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Protozoan Parasite ◽  
18S Rrna Gene ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Cryptosporidium Spp ◽  
Potential Source ◽  
Wide Range ◽  
First Time

Abstract Cryptosporidium is a protozoan parasite with a wide range of hosts, including humans. However, only a few Cryptosporidium species have been described in birds (C. meleagridis, C. baileyi, C. galli and C. avium). The aim of this study was to investigate the occurrence of Cryptosporidium spp. in feces of eared doves (Zenaida auriculata), followed by molecular characterization of the parasite. A total of 196 animals of both sexes were trap-captured; the animals were culled and the intestinal contents were collected for DNA extraction. After extraction, a nested-PCR (nPCR), which amplifies a fragment of the 18S rRNA gene of Cryptosporidium spp., was performed. The amplicons obtained were purified and sequenced. PCR analysis revealed that 30 animals (15.3%) were positive for Cryptosporidium spp. There was no significant sex-dependent enrichment of Cryptosporidium occurrence (p > 0.05). Only 15 out of the 30 positive samples were successfully sequenced and their species determined, of which, 13 (86.7%) and 2 (13.3%) were C. meleagridis and C. galli, respectively. Herein, we present for the first time a molecular characterization of Cryptosporidium from feces of eared doves (Z. auriculata) and propose that these birds are a potential source of C. meleagridis infection in humans.

scholarly journals Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil

2017 ◽  
Vol 37 (12) ◽  
pp. 1416-1422 ◽  
Author(s):  
Rita de Maria Seabra Nogueira ◽  
Arannadia Barbosa Silva ◽  
Tayra Pereira Sato ◽  
Joicy Cortez de Sá ◽  
Ana Clara Gomes dos Santos ◽  
...  
Keyword(s):  
Anaplasma Phagocytophilum ◽  
Animal Health ◽  
Antibody Test ◽  
18S Rrna Gene ◽  
Boophilus Microplus ◽  
Rrna Gene ◽  
Serum Samples ◽  
Theileria Equi ◽  
Babesia Caballi ◽  
Blood Samples

ABSTRACT: Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT) and blood samples and ticks to Polymerase Chain Reaction (PCR) to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. It was detected 2.35% (4/170) and 0.59% (1/170) positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05) was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.

scholarly journals Molecular evaluation of piroplasms and hematological changes in canine blood stored in a clinical laboratory in Niterói, Rio de Janeiro

2020 ◽  
Vol 29 (3) ◽  
Author(s):  
Fernanda Barbosa dos Santos ◽  
Gilberto Salles Gazeta ◽  
Laís Lisboa Correa ◽  
Lucas Fernandes Lobão ◽  
João Pedro Palmer ◽  
...  
Keyword(s):  
Rio De Janeiro ◽  
Clinical Laboratory ◽  
18S Rrna Gene ◽  
Metropolitan Region ◽  
Rrna Gene ◽  
Babesia Vogeli ◽  
Hematological Changes ◽  
Pcr Targeting ◽  
Canine Piroplasmosis

Abstract Piroplasm species were analyzed by molecular tools in total 31 blood samples from positive dogs, previously checked by stained slides, stored until DNA extraction between 2016 to 2018 in the laboratory Clinical Analyzes in Niterói, Rio de Janeiro. The piroplasms were identified by PCR, targeting the 18S rRNA gene and sequencing. From the total number of samples only 24 (77.4%) were positive and show adequate nucleotide sequences for interpretation with identity between 93%-100% with Babesia vogeli in compared to the sequences isolated of infected dogs from other states in Brazil deposited on GenBank. Most of dogs infected with B. vogeli had anemia (62.5%) and thrombocytopenia (95.8%). The findings of this study are compatible with previous reports in the literature and highlight B. vogeli as the most incriminated species in canine piroplasmosis in Brazil, and thrombocytopenia the hematological alteration most frequently identified in this infection. It is important to note that this is the first study involving the molecular characterization of piroplasms in the metropolitan region of Rio de Janeiro, based on PCR followed by sequencing.

TBE in Sweden

2020 ◽  
Keyword(s):  
Genetic Characterization ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Second Phase ◽  
Serum Samples ◽  
Tick Borne Encephalitis ◽  
Specific Immunoglobulin ◽  
Tick Borne Encephalitis Virus ◽  
First Time

Tick-borne encephalitis virus (TBEV) was isolated for the first time in Sweden in 1958 (from ticks and from 1 tick-borne encephalitis [TBE] patient).1 In 2003, Haglund and colleagues reported the isolation and antigenic and genetic characterization of 14 TBEV strains from Swedish patients (samples collected 1991–1994).2 The first serum sample, from which TBEV was isolated, was obtained 2–10 days after onset of disease and found to be negative for anti-TBEV immunoglobulin M (IgM) by enzyme-linked immunosorbent assay (ELISA), whereas TBEV-specific IgM (and TBEV-specific immunoglobulin G/cerebrospinal fluid [IgG/CSF] activity) was demonstrated in later serum samples taken during the second phase of the disease.

scholarly journals Molecular Characterization of Ciliate Diversity in Stream Biofilms

2008 ◽  
Vol 74 (6) ◽  
pp. 1740-1747 ◽  
Author(s):  
Andrew Dopheide ◽  
Gavin Lear ◽  
Rebecca Stott ◽  
Gillian Lewis
Keyword(s):  
18S Rrna ◽  
Pcr Primers ◽  
18S Rrna Gene ◽  
Sequence Diversity ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Sequencing Analysis ◽  
Specific Pcr ◽  
Stream Biofilms

ABSTRACT Free-living protozoa are thought to be of fundamental importance in aquatic ecosystems, but there is limited understanding of their diversity and ecological role, particularly in surface-associated communities such as biofilms. Existing eukaryote-specific PCR primers were used to survey 18S rRNA gene sequence diversity in stream biofilms but poorly revealed protozoan diversity, demonstrating a need for protozoan-targeted primers. Group-specific PCR primers targeting 18S rRNA genes of the protozoan phylum Ciliophora were therefore designed and tested using DNA extracted from cultured protozoan isolates. The two most reliable primer combinations were applied to stream biofilm DNA, followed by cloning and sequencing analysis. Of 44 clones derived from primer set 384F/1147R, 86% were of probable ciliate origin, as were 25% of 44 clones detected by primer set 121F/1147R. A further 29% of 121F/1147R-detected clones matched sequences from the closely related phylum Apicomplexa. The highly ciliate-specific primer set 384F/1147R was subsequently used in PCRs on biofilm DNA from four streams exhibiting different levels of human impact, revealing differences in ciliate sequence diversity in samples from each site. Of a total of 240 clones, 73% were of probable ciliate origin; 54 different putative ciliate sequences were detected from throughout seven taxonomic ciliate classes. Sequences from Oligohymenophorea were most commonly detected in all samples, followed by either Spirotrichea or Phyllopharyngea. Restriction fragment length polymorphism profile-based analysis of clones suggested a potentially higher level of diversity than did sequencing. Nevertheless, newly designed PCR primers 384F/1147R were considered to provide an effective molecular basis for characterization of ciliate diversity in stream biofilms.

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