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 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 ◽  
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 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 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 First Molecular Detection and Characterization of Hemotropic Mycoplasma Species in Cattle and Goats from Uganda

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1624
Author(s):  
Benedicto Byamukama ◽  
Maria Agnes Tumwebaze ◽  
Dickson Stuart Tayebwa ◽  
Joseph Byaruhanga ◽  
Martin Kamilo Angwe ◽  
...  
Keyword(s):  
Mammalian Species ◽  
Mycoplasma Species ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
Sequencing Analysis ◽  
East African ◽  
African Region ◽  
Mycoplasma Wenyonii ◽  
East African Region ◽  
Western Uganda

Hemoplasmas (hemotropic mycoplasmas) are small pleomorphic bacteria that parasitize the surface of red blood cells of several mammalian species including cattle, goats, and humans, causing infectious anemia. However, studies on hemoplasmas have been neglected and to date, there are no studies on bovine and caprine hemoplasmas in Uganda or the entire East African region. In this study, a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene was used to investigate the presence of hemoplasma in 409 samples (cattle = 208; goats = 201) collected from Kasese district, western Uganda. Results showed that 32.2% (67/208) of cattle samples and 43.8% (88/201) of goat samples were positive for hemoplasmas. Sequencing analysis identified Candidatus Mycoplasma haemobos and Mycoplasma wenyonii in cattle, while Candidatus Mycoplasma erythrocervae and Mycoplasma ovis were identified in goats. Statistical analysis showed that goats were at a higher risk of infection with hemoplasmas compared with cattle. To the best of our knowledge, this is the first molecular evidence of hemoplasmas in bovine and caprine animals in Uganda and the entire east African region.

scholarly journals Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I

2017 ◽  
Vol 26 (2) ◽  
pp. 177-184 ◽  
Author(s):  
Hilda Fátima de Jesus Pena ◽  
Camila Mariellen Evangelista ◽  
Renata Assis Casagrande ◽  
Giovana Biezus ◽  
Claudia Salete Wisser ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Histopathological Examination ◽  
Leukemia Virus ◽  
Fatal Case ◽  
Domestic Cat ◽  
Feline Leukemia Virus ◽  
Type I ◽  
Clonal Type ◽  
Pcr Rflp ◽  
Pcr Targeting

Abstract The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV). A domestic cat (Felis silvestris catus) presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC) evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

scholarly journals First report and molecular characterization of Cryptosporidium spp. in humans and animals in Khartoum state, Sudan

2019 ◽  
Vol 12 (1) ◽  
pp. 183-189 ◽  
Author(s):  
Kaltoum Yagoub Adam ◽  
A. A. Ismail ◽  
M. A. Masri ◽  
A. A. Gameel
Keyword(s):  
Nested Pcr ◽  
Mammalian Species ◽  
Pcr Amplification ◽  
Health Concern ◽  
Economic Losses ◽  
Staining Procedure ◽  
Rrna Gene ◽  
Nested Polymerase Chain Reaction ◽  
Cryptosporidium Spp ◽  
Pcr Targeting

Background and Aim: Cryptosporidium is recognized to infect several mammalian species as well as humans, causing substantial economic losses and serious public health concern. Infected animals can be a source of environmental contamination and human infections. In general, the occurrence of Cryptosporidium species in animals and human in Sudan and zoonotic importance is not well documented. This study aimed to identify Cryptosporidium spp. infecting different animal species and humans and to compare between different isolates obtained. Materials and Methods: To provide molecular information about Cryptosporidium in animals and humans, both modified Ziehl-Neelsen (MZN) specific stain and molecular assay were used. Concentration techniques followed by three protocols of DNA extraction were carried out. After microscopic screening of 263 fecal samples (goats [n=197], cattle [n=12], sheep [n=12], and human [n=42]), 61 positive and 30 negative, randomly selected samples were used in nested polymerase chain reaction (PCR) targeting part of the 18S RNA. Results: Nested PCR amplification confirmed 91.8% (56/61) of microscopic-positive samples. 8.2% (5/61) of negative samples by PCR (positive by microscopy) were considered false negatives. Sequencing followed by alignment of the 14 isolates indicated that all samples were identical (100%) and belonged to Cryptosporidium parvum. Conclusion: MZN staining procedure is reliable for the routine diagnosis of Cryptosporidium; cetyltrimethylammonium bromide extraction buffer and nested PCR targeting 18S rRNA gene are reliable and useful in epidemiological studies of this parasite.

scholarly journals Molecular Evidence of Chlamydia-Like Organisms in the Feces of Myotis daubentonii Bats

2016 ◽  
Vol 83 (2) ◽  
Author(s):  
K. Hokynar ◽  
E. J. Vesterinen ◽  
T. M. Lilley ◽  
A. T. Pulliainen ◽  
S. J. Korhonen ◽  
...  
Keyword(s):  
16S Rrna ◽  
Sequence Similarity ◽  
Pcr Amplification ◽  
Rrna Genes ◽  
23S Rrna ◽  
Molecular Evidence ◽  
Rrna Gene ◽  
16S Rrna Sequence ◽  
Content Type ◽  
Pcr Targeting

ABSTRACT Chlamydia-like organisms (CLOs) are recently identified members of the Chlamydiales order. CLOs share intracellular lifestyles and biphasic developmental cycles, and they have been detected in environmental samples as well as in various hosts such as amoebae and arthropods. In this study, we screened bat feces for the presence of CLOs by molecular analysis. Using pan-Chlamydiales PCR targeting the 16S rRNA gene, Chlamydiales DNA was detected in 54% of the specimens. PCR amplification, sequencing, and phylogenetic analysis of the 16S rRNA and 23S rRNA genes were used to classify positive specimens and infer their phylogenetic relationships. Most sequences matched best with Rhabdochlamydia species or uncultured Chlamydia sequences identified in ticks. Another set of sequences matched best with sequences of the Chlamydia genus or uncultured Chlamydiales from snakes. To gain evidence of whether CLOs in bat feces are merely diet borne, we analyzed insects trapped from the same location where the bats foraged. Interestingly, the CLO sequences resembling Rhabdochlamydia spp. were detected in insect material as well, but the other set of CLO sequences was not, suggesting that this set might not originate from prey. Thus, bats represent another potential host for Chlamydiales and could harbor novel, previously unidentified members of this order. IMPORTANCE Several pathogenic viruses are known to colonize bats, and recent analyses indicate that bats are also reservoir hosts for bacterial genera. Chlamydia-like organisms (CLOs) have been detected in several animal species. CLOs have high 16S rRNA sequence similarity to Chlamydiaceae and exhibit similar intracellular lifestyles and biphasic developmental cycles. Our study describes the frequent occurrence of CLO DNA in bat feces, suggesting an expanding host species spectrum for the Chlamydiales. As bats can acquire various infectious agents through their diet, prey insects were also studied. We identified CLO sequences in bats that matched best with sequences in prey insects but also CLO sequences not detected in prey insects. This suggests that a portion of CLO DNA present in bat feces is not prey borne. Furthermore, some sequences from bat droppings not originating from their diet might well represent novel, previously unidentified members of the Chlamydiales order.

scholarly journals Long-Range Effects of Retroviral Insertion on c-myb: Overexpression May Be Obscured by Silencing during Tumor Growth In Vitro

2003 ◽  
Vol 77 (2) ◽  
pp. 1059-1068 ◽  
Author(s):  
L. Hanlon ◽  
N. I. Barr ◽  
K. Blyth ◽  
M. Stewart ◽  
P. Haviernik ◽  
...  
Keyword(s):  
T Cell ◽  
Long Range ◽  
Cell Lines ◽  
Leukemia Virus ◽  
Mammalian Species ◽  
Transcription Unit ◽  
Feline Leukemia Virus ◽  
Expression Levels

ABSTRACT The c-myb oncogene is a frequent target for retroviral activation in hemopoietic tumors of avian and mammalian species. While insertions can target the gene directly, numerous clusters of retroviral insertion sites have been identified which map close to c-myb and outside the transcription unit in T-lymphomas (Ahi-1, fit-1, and Mis-2) and monocytic and myeloid leukemias (Mml1, Mml2, Mml3, and Epi-1). Previous analyses showed no consistent effect of these insertions on c-myb expression, raising the possibility that other nearby genes were the true targets. In contrast, our analysis of four cell lines established from lymphomas bearing insertions at fit-1 (fti-1) (feline leukemia virus) and Ahi-1 (Moloney murine leukemia virus) shows that these display higher expression levels of c-myb RNA and protein compared to a panel of phenotypically similar cell lines lacking such insertions. An interesting feature of the cell lines with long-range c-myb insertions was that each also carried an activated Myc allele. The potential for oncogenic synergy between Myb and Myc in T-cell lymphoma was confirmed in transgenic mice overexpressing alleles of both genes in the T-cell compartment, lending further credence to the case for c-myb as the major target for long-range activation. In contrast, mapping and analysis of c-myb neighboring genes (HBS1 and FLJ20069) showed that the expression of these genes did not correlate well with the presence of proviral insertions. A possible explanation for the paradoxical behavior of c-myb was provided by one of the murine T-lymphoma lines bearing an insertion at Ahi-1 (p/m16i) that reproducibly down-regulated c-myb RNA and protein to very low levels or undetectable levels on prolonged culture. Our observations implicate c-myb as a key target of upstream and downstream retroviral insertions. However, overexpression may become dispensable during outgrowth in vitro, and perhaps during tumor progression in vivo, providing a potential rationale for the previously observed discordance between retroviral insertion and c-myb expression levels.

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