scholarly journals Molecular confirmation of hemothropic mycoplasmas (hemoplasmas) in domestic cats in Romania

2020 ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Tijana Suici ◽  
...  
Keyword(s):  
Mammalian Species ◽  
Small Animal ◽  
Conventional Polymerase Chain Reaction ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Candidatus Mycoplasma Turicensis ◽  
Sampling Season

Abstract Background The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemothropic mycoplasmas in client owned cats in Romania. Methods Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. Results Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, FeLV/FIV positivity of cats or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Conclusions The findings support the emergence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

scholarly journals Molecular confirmation of hemothropic mycoplasmas (hemoplasmas) in domestic cats (Felis catus) in Romania

2020 ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Viorel Herman ◽  
...  
Keyword(s):  
Mammalian Species ◽  
Small Animal ◽  
Conventional Polymerase Chain Reaction ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Candidatus Mycoplasma Turicensis ◽  
Sampling Season

Abstract Background: The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemothropic mycoplasmas in client owned cats in Romania. Methods : Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. Results: Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0 . 05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, FeLV/FIV positivity of cats or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Conclusions : The findings support the emergence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

scholarly journals Molecular detection of hemotropic mycoplasmas (hemoplasmas) in domestic cats (Felis catus) in Romania

2020 ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Viorel Herman ◽  
...  
Keyword(s):  
Population Genetic ◽  
Leukemia Virus ◽  
Mammalian Species ◽  
Small Animal ◽  
Conventional Polymerase Chain Reaction ◽  
Feline Leukemia Virus ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Immunodeficiency Virus

Abstract Background: The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemotropic mycoplasmas in client-owned cats in Romania.Methods: Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction targeting the 16S rRNA gene and sequencing assays. PCR-positive samples were subsequently analyzed by phylogenetic and population genetic analysis.Results: Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations (p > 0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, feline leukemia virus/feline immunodeficiency virus (FeLV/FIV) positivity of cats, or the sampling season. However, outdoor access was positively associated (p=0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Phylogenetic analysis revealed that our sequences clustered with those selected from the GenBank database in two distinct clades. The registered population genetic indices were strongly supportive of the great variance in sequences between the recorded Mycoplasma species.Conclusions: The findings support the occurrence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

scholarly journals Molecular detection of hemotropic mycoplasmas (hemoplasmas) in domestic cats (Felis catus) in Romania

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Viorel Herman ◽  
...  
Keyword(s):  
Population Genetic ◽  
Leukemia Virus ◽  
Mammalian Species ◽  
Small Animal ◽  
Conventional Polymerase Chain Reaction ◽  
Feline Leukemia Virus ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Population Genetic Analysis ◽  
Pcr Targeting

Abstract Background The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemotropic mycoplasmas in client-owned cats in Romania. Methods Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. PCR-positive samples were subsequently analyzed by phylogenetic and population genetic analysis. Results Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations (p > 0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, feline leukemia virus/feline immunodeficiency virus positivity of cats, or the sampling season. However, outdoor access was positively associated (p = 0.049) with infection and could be considered a risk factor (OR = 4.1) in acquiring feline hemotropic mycoplasmas. Phylogenetic analysis revealed that our sequences clustered with those selected from the GenBank database in two distinct clades. The registered population genetic indices were strongly supportive of the great variance in sequences between the recorded Mycoplasma species. Conclusions The findings support the occurrence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

scholarly journals Molecular detection of hemotropic mycoplasmas (hemoplasmas) in domestic cats (Felis catus) in Romania

2020 ◽  
Author(s):  
Mirela Imre ◽  
Cristina Văduva ◽  
Gheorghe Dărăbuș ◽  
Sorin Morariu ◽  
Viorel Herman ◽  
...  
Keyword(s):  
Population Genetic ◽  
Leukemia Virus ◽  
Mammalian Species ◽  
Small Animal ◽  
Feline Leukemia Virus ◽  
Mycoplasma Species ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Population Genetic Analysis ◽  
Pcr Targeting

Abstract Background: The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemotropic mycoplasmas in client-owned cats in Romania. Methods : Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. PCR-positive samples were subsequently analyzed by phylogenetic and population genetic analysis. Results: Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0 . 05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, feline leukemia virus/feline immunodeficiency virus (FeLV/FIV) positivity of cats, or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Phylogenetic analysis revealed that our sequences clustered with those selected from the GenBank database in two distinct clades. The registered population genetic indices were strongly supportive of the great variance in sequences between the recorded Mycoplasma species. Conclusions : The findings support the occurrence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

scholarly journals Genetic diversity of Ehrlichia canisstrains from naturally infected dogs in Rio de Janeiro, Brazil

2014 ◽  
Vol 23 (3) ◽  
pp. 301-308 ◽  
Author(s):  
Renata Fernandes Ferreira ◽  
Aloysio de Mello Figueiredo Cerqueira ◽  
Tatiana Xavier de Castro ◽  
Eliane de Oliveira Ferreira ◽  
Felipe Piedade Gonçalves Neves ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Rio De Janeiro ◽  
Complete Sequence ◽  
Rrna Gene ◽  
Hybrid Strain ◽  
The Us ◽  
Polymerase Chain ◽  
Hematological Manifestations ◽  
Pcr Targeting ◽  
The 16S Rrna Gene

The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of 1the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.

scholarly journals Prevalence and genetic characterization of Toxoplasma gondii in naturally infected backyard pigs intended for familial consumption in Romania

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Anamaria Ioana Paştiu ◽  
Anamaria Cozma-Petruț ◽  
Aurélien Mercier ◽  
Anamaria Balea ◽  
Lokman Galal ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rural Areas ◽  
Antibody Test ◽  
Antibody Detection ◽  
Serum Samples ◽  
Potential Threat ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Backyard Pigs

Abstract Background Foodborne toxoplasmosis in humans can be due to the exposure to tissue cysts of Toxoplasma gondii through the consumption of meat, including pork, of infected animals. Traditional Romanian food habits include pork as the preferred meat, while backyard pig rearing remains a common practice in many rural areas of Romania. The aims of the present study were to estimate the prevalence of T. gondii infection in naturally infected backyard pigs slaughtered for familial consumption and to genetically characterize the T. gondii strains obtained. Methods Paired blood and heart samples were collected from 94 backyard pigs, home slaughtered for private consumption. Serum samples were analyzed using the immunofluorescence antibody test (IFAT) for anti-T. gondii antibody detection. Heart samples were screened by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529) for T. gondii detection. In addition, heart samples from IFAT positive animals were bioassayed in mice. The T. gondii isolates were genotyped by the analysis of 15 microsatellite markers. Results The results showed that almost half of the pigs investigated were T. gondii seropositive (46.8%, 95% confidence interval (CI): 36.4–57.4%) and in more than a quarter of the pigs (26.6%, 95% CI: 18.0–36.7%), the parasite was detected by PCR. Three (3/44) T. gondii strains were isolated from hearts of seropositive pigs and they all belonged to genotype II. Conclusions The present study showed the presence of T. gondii infection in backyard pigs in Romania, which suggests that consumption of pork from animals reared and slaughtered at home may pose a potential threat to human health and should be given attention. In addition, to our knowledge, this is the first study to provide data concerning T. gondii strains circulating in pigs from Romania.

scholarly journals Hematological values associated to the serological and molecular diagnostic in cats suspected of Ehrlichia canisinfection

2013 ◽  
Vol 22 (4) ◽  
pp. 470-474 ◽  
Author(s):  
ísis Assis Braga ◽  
Luana Gabriela Ferreira dos Santos ◽  
Andréia Lima Tomé Melo ◽  
Felipe Wolf Jaune ◽  
Thaysa Felfili Ziliani ◽  
...  
Keyword(s):  
Molecular Diagnostic ◽  
Rrna Gene ◽  
Igg Antibody ◽  
Mato Grosso ◽  
Pcr Products ◽  
Antibody Titers ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Veterinary Hospital ◽  
Hematological Alterations

The literature contains several studies on feline ehrlichiosis. However, information about the characteristics of Ehrlichiainfection in cats is still scanty. This study evaluated the association between Ehrlichia spp. infection and the hematologic data of 93 cats treated at the Federal University of Mato Grosso Veterinary Hospital in Cuiabá, state of Mato Grosso, Brazil. The presence of or exposure to Ehrlichia spp. infection was evaluated by Polymerase Chain Reaction (PCR) targeting the dsb and 16S rRNA gene of Ehrlichia, and by detection of anti-Ehrlichia canis IgG antibodies in Indirect Fluorescence Assay (IFA), respectively. Eight (8.6%) cats tested positive by PCR and the partial DNA sequence obtained from PCR products was a 100% match to E. canis. Forty-two (45.1%) cats showed antibody reactivity against Ehrlichia spp. Hematological alterations such as low erythrocyte count, thrombocytopenia, lymphopenia and monocytosis were observed in PCR positive cats. Among them, low erythrocyte counts were associated with IgG antibody titers of 40 to 640 and five cats also tested positive by PCR. Furthermore, PCR-positive cats showed a tendency to be lymphopenic. No correlation was found between age and sex, and no ticks were observed in any of the examined cats.

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 Descriptive Investigation of Strongyloidiasis Infection and Characterization of Strongyloides stercoralis Using Morphological and Molecular-Based Methods

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Nayana Gunathilaka ◽  
Nilmini Chandrasena ◽  
Tharaka Wijerathna ◽  
Yoshito Fuji ◽  
Deepa Gunasekara ◽  
...  
Keyword(s):  
Gastrointestinal Symptoms ◽  
Strongyloides Stercoralis ◽  
Molecular Evidence ◽  
Pcr Assay ◽  
Its1 Region ◽  
Promising Tool ◽  
Hyperinfection Syndrome ◽  
Polymerase Chain ◽  
Stool Cultures

Strongyloidiasis is caused by the nematode Strongyloides stercoralis which has the unique ability to reproduce and complete its entire life cycle within the human host through its autoinfection cycle. Diagnosis of this infection is important because of its potential to cause fatal hyperinfection syndrome or disseminated infections in those with defective cellular immunity. Parasitological methods based on faecal microscopy and culture often fail to detect low-intensity infections. A multiplex polymerase chain reaction (PCR) assay was developed for the detection of S. stercoralis, Ascaris lumbricoides, and Enterobius vermicularis by designing primers specific for the ITS1 region of ribosomal DNA of S. stercoralis and A. lumbricoides and 18S region of rRNA of E. vermicularis. A 61-year-old patient presented with chronic gastrointestinal and respiratory symptoms and weight loss with a stool microscopy positive for helminth larvae. Stool cultures with the Harada–Mori technique yielded L3 larvae which were identified as S. stercoralis based on morphology. The multiplex PCR performed on DNA extracted from stool elicited the expected band at 129 bp on gel electrophoresis of the PCR yield providing molecular evidence of intestinal strongyloidiasis. The patient’s gastrointestinal symptoms improved with a six-day course of albendazole (400 mg twice daily). Negative posttreatment stool microscopy, culture, and PCR confirmed successful clearance of infection. Molecular-based PCR assay is a promising tool to diagnose and assess the therapeutic efficacy of anthelmintics in intestinal helminthiases such as strongyloidiasis.

scholarly journals Molecular analysis of Blastocystis sp. and its subtypes from treated wastewater routinely used for irrigation of vegetable farmlands in Iran

2019 ◽  
Vol 17 (5) ◽  
pp. 837-844 ◽  
Author(s):  
Ehsan Javanmard ◽  
Hanieh Mohammad Rahimi ◽  
Maryam Niyyati ◽  
Hamid Asadzadeh Aghdaei ◽  
Meysam Sharifdini ◽  
...  
Keyword(s):  
Potential Role ◽  
Treated Wastewater ◽  
Rrna Gene ◽  
Pathogenic Microorganisms ◽  
Chain Reaction ◽  
Blastocystis Sp ◽  
Polymerase Chain ◽  
Wastewater Samples ◽  
Pcr Targeting

Abstract Treated wastewater samples were collected, filtered using sterile 47-mm cellulose nitrate membrane and DNA extracted from the filtered materials. The presence of Blastocystis sp. was confirmed via polymerase chain reaction (PCR) targeting the SSU rRNA gene of Blastocystis sp. in 5/12 of samples. Based on the subtype analysis after sequencing, 2, 2 and 1 of ST2, ST6 and ST8 were detected among the isolates, respectively. Furthermore, both ST6s were allele 139, alleles 11 and 138 were identified in ST2 and the only ST8 was allele 95. The phylogenetic tree showed that one of ST2 was clustered together with those ST2 that were already reported from humans and animals. The presence of Blastocystis sp. in treated wastewater can indicate the potential role of this type of water for irrigation in the transmission of pathogenic microorganisms to downstream farmlands.

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