Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Bacanelli, Gisele; Ramos, Carlos A. N.; Araújo, Flábio R.

doi:10.1590/s0100-736x2014000100005

Cited by 9 publications

(12 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…marginale. This endemic stability for anaplasmosis was similarly reported in a previous study in Brazil (Bacanelli et al, 2014). Endemic stability is an epidemiological state, in which there is high haemoparasites-infected ticks but clinical disease is absent in the population (Jonsson et al, 2012).…”

Section: Discussionsupporting

confidence: 85%

“…Although conventional PCR has been shown to be useful to determine the presence of Babesia and Anaplasma (Figueroa et al, 1993;Martins et al, 2008;Bilgiç et al, 2013;El-Ashker et al, 2015), more recently, qPCR has revealed to be more sensitive than the first for the detection of protozoan parasites in cattle (Criado-Fornelio et al, 2009). For the detection of anaplasmosis, some of the TaqMan ® based qPCR methods involve the amplification of species-specific genes such as the major surface proteins (msp) genes (Carelli et al, 2007;Bacanelli et al, 2014), and six msp (msp 1a, 1b, 2, 3, 4, 5) with the potential to be used in diagnostic assays have been identified on A. marginale (Kocan et al, 2003). The msp1 gene complex (composed of msp1a and msp1b) has been shown to be a well-conserved gene in A. marginale (de la Fuente et al, 2001;Bowie et al, 2002), whose products act as adhesin during invasion of bovine erythrocytes (McGarey et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

“…In the present study, the TaqMan ® qPCR used were highly specific, and their sensitivity has been previously evaluated for A. marginale (Carelli et al, 2007). Probe-based qPCR methods are more expensive but, when compared to other PCR methods have multiple advantages, including the detection of very low infections, no requirement of post PCR steps, and they are a useful quantitative technique for chemotherapeutic and vaccine evaluation ( Monis et al, 2005;Bacanelli et al, 2014). They are also useful in processing several samples simultaneously for large scale epidemiological studies (Carelli et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR

et al. 2016

View full text Add to dashboard Cite

Tick-borne diseases (TBDs) are some of the most important animal health and management problems in Africa, including Nigeria. This study aims to determine the prevalence of an important TBD, anaplasmosis, in a North-central region of Nigeria. Blood samples were collected from cattle and stored on Whatman FTA cards. Information on village, age and sex associated with each cattle was also recorded. The packed red blood cell volume (PCV) for each blood sample was determined. After DNA extraction, pathogen presence was evaluated by TaqMan based qPCR of which 75.9 % of the cattle tested positive for Anaplasma marginale. Statistical analysis revealed that the presence of A. marginale infection differed significantly between cattle age groups. However, there was no significant difference in the prevalence of this pathogen between the sexes or among cattle grouped by PCV level. Finally, using a highly sensitive molecular method our pioneer study contributes to the improvement of the current knowledge regarding tick-borne pathogens that seriously affect animal health in specific areas of Nigeria.

show abstract

Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR

et al. 2016

View full text Add to dashboard Cite

show abstract

“…In the comparative analysis of the snPCR result for msp5 gene among splenectomized animals, cattle presented a significantly higher average frequency of infection than buffaloes. Other studies also reported that buffaloes had lower values (Silva et al 2014a(Silva et al , 2014b compared to the observed in cattle (Bacanelli et al 2014). A splenectomized buffalo was positive only 60 days after inoculation.…”

Section: Discussionmentioning

confidence: 78%

“…Two hypotheses may support this. The first, the animals came into contact with native strains of A. marginale from the moment they went to pasture, which contributed to a greater detection in snPCR for msp5 gene, present in several Anaplasma species and used for serological and molecular screening studies (Torioni de Echaide et al 1998, Bacanelli et al 2014. The second, over time, the AmRio 2 strain increased the number of DNA copies in the circulatory current of the ruminants and, consequently, contributed to a higher sensitivity of the molecular examination and this can be supported by the fact that this time (pasture period) includes the period when rickettsemia increases (2 to 6 weeks after infection), in which levels of 10 9 or more bacteria per milliliter of blood can be verified by quantitative PCR (Futse et al 2003, Han et al 2010.…”

Section: Discussionmentioning

confidence: 99%

Experimental infection by Anaplasma marginale in buffaloes and cattle: clinical, hematological, molecular and pathological aspects

et al. 2019

View full text Add to dashboard Cite

The study aimed to evaluate and compare the clinical, laboratory and pathological aspects of buffalo and bovine experimentally infected with AmRio 2 strain of Anaplasma marginale. Four Murrah buffaloes and four crossbred cattle were used in the experiment, which two animals of each species were splenectomized. Strain AmRio 2 of A. marginale was inoculated in all experimental animals. Clinical exams, Packed Cell Volume (PCV), blood counts, blood smears, rickettsemia, necropsy and histopathology were performed in all cases. Semi-Nested-PCR (snPCR) for the msp5 and snPCR for the msp1α target gene for identification of A. marginale in blood samples from animals was done. From positive samples for msp1α snPCR, samples were analyzed for the amino acid sequences of this gene. Two splenectomized cattle presented apathy, pale mucous membranes, jaundice, hyperthermia, and severe anemia. The remaining experimental animals did not show clinical signs. The rickettsemia in all animals was less than 1%. The mean PCV of the splenectomized cattle was below 20% at two-time points after infection. On the blood count, the main changes were observed in splenectomized calves and were characterized by a decrease in red blood cells, hemoglobin, PCV and platelets (p <0.05). All animals presented leukocyte elevation by increased lymphocytes, however, with no significant difference. The average prepatent period was two days in all the animals. The average incubation period in cattle that became ill was 25.5 days, and death occurred, on average, 63 days after inoculation of the strain. The necropsy findings were characterized by pale carcass, ascites, enlarged liver, distended gallbladder, and thick bile. Histopathological findings included infiltration of macrophages and lymphocytes in various organs, hepatic sinusoidal dilatation, and necrosis of the large intestine. In snPCR for the msp5 gene, 100% of the animals were positive in at least one evaluation. And in the snPCR for the infection of the msp1α target gene was also found in all animals in at least one sample evaluated. However, sequencing revealed only five animals, including the bovine which died, with a similarity of the amino acid sequences with AmRio 2 strain of A. marginale. It is concluded that the splenectomized cattle died due to anaplasmosis caused by the inoculated strain and the buffalo were more resistant compared to cattle. Buffaloes can be an alternative to cattle rearing in areas with a high occurrence of clinical cases of anaplasmosis.

show abstract

A study of biochemical changes in river buffaloes (Bubalus bubalis) naturally infected with Anaplasma marginale

Jalali,

Nikvand,

Gharibi

et al. 2023

J Hellenic Vet Med Soc

View full text Add to dashboard Cite

Anaplasma marginale as an intracellular rickettsia, endemic in tropical and subtropical regions of the world. This study was performed to determine A. marginale infection, molecularly, and to investigate its effects on some serum biochemical parameters in river buffaloes referred to Ahvaz abattoir, Southwest Iran. A total of 103 apparently healthy river buffaloes were randomly sampled. Whole blood samples were subjected to PCR analysis and blood smears were examined microscopically for the presence of Anaplasma inclusions. Serum biochemical parameters including total protein, albumin, glucose, urea, iron, total bilirubin, total cholesterol, Calcium, Phosphorus and Magnesium concentrations, aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activities and also serum electrolytes comprising Na and K were assessed. The results revealed that 16 and 32 samples were infected with A. marginale in microscopic and PCR assessment, respectively. Serum biochemical analysis showed no significant difference between A. marginale infected and non-infected buffalo groups. It can be concluded that infection with A. marginale does not lead to much pathogenicity in river buffaloes, so that in infected animals, no clinical signs or abnormalities in the performance indicators of various organs can be detected at the laboratory level. However, the role of this species as a reservoir of this microorganism in nature and in the epidemiology and transmission of anaplasmosis to other species remains significant. Buffaloes can be a substitute to raising cattle in regions with high occurrence of anaplasmosis.

show abstract

Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Cited by 9 publications

References 20 publications

Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR

Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR

Experimental infection by Anaplasma marginale in buffaloes and cattle: clinical, hematological, molecular and pathological aspects

A study of biochemical changes in river buffaloes (Bubalus bubalis) naturally infected with Anaplasma marginale

Contact Info

Product

Resources

About