Molecular detection of Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos, Trypanosoma evansi and first evidence of Theileria sinensis-associated bovine anaemia in crossbred Kedah-Kelantan x Brahman cattle

Abstract Background Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos and Trypanosoma evansi, and determine their phylogenetic relationships and haemato-biochemical abnormalities in naturally infected cattle. Methods Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, MPSP genes of T. orientalis and T. sinensis, MSP4 gene of A. marginale, 16SrRNA gene of Candidatus Mycoplasma haemobos, and RoTat1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) clinically ill Kedah-Kelantan x Brahman cattle in Pahang, Malaysia. Results A total of 44 (72.13%) cattle were infected with more than one blood pathogen. Theileria species was the blood pathogen with the highest molecular detection rate (72.13, 95% CI 59.83–81.81%). Nucleotide blast analyses of all sequences demonstrated high degree of molecular similarity (98–100%) in comparison with their respective reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16SrRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys respectively as additional species detected in these cattle. MPSP-PCR analysis was conducted for further confirmation of T. sinensis. The blood picture of eight infected cattle groups revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. High mean erythrocyte fragility values were common in infected cattle groups. Anaemia of the macrocytic normochromic type and spherocytes were observed in the T. evansi and Anaplasma platys + Theileria sinensis double species co-infected cattle group. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (p < 0.05) increases in serum liver and kidney parameters, total protein, globulin, total and unconjugated bilirubin and decreased albumin values were observed in the T. evansi infected cattle when compared to clinically healthy cattle. Conclusion We present the first evidence of Theileria sinensis-associated bovine anaemia (TSABA) in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of A. platys, there is an urgent need for appropriate preventive and control measures against these blood pathogens.

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First report of bovine anaemia associated Theileria sinensis infection and phylogenetic analyses of partial gene sequences of Theileria and Anaplasma species detected in naturally infected Malaysian cattle

10.21203/rs.3.rs-17415/v1 ◽

2020 ◽

Cited By ~ 1

Author(s):

Onyinyechukwu Ada Agina ◽

Mohd Rosly Shaari ◽

Nur Mahiza Isa ◽

Mokrish Ajat ◽

Mohd Zamri-Saad ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Pcr Amplification ◽

Trypanosoma Evansi ◽

Control Measures ◽

Rrna Gene ◽

Infected Cattle ◽

Anaplasma Platys ◽

Gene Sequences ◽

First Report

Abstract Background: Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Trypanosoma evansi, Theileria, Anaplasma and Mycoplasma species, and studied their phylogenetic relationships, haemato-biochemical abnormalities and erythrocyte osmotic fragility (EOF) in Malaysian cattle. Methods: Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, 16SrRNA gene of Candidatus Mycoplasma haemobos, MPSP gene of T. orientalis and RoTaT1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) Kedah-Kelantan X Brahman cattle from Pahang, Malaysia. Haemato-biochemical analyses were performed using automated analysers while EOF was determined with the aid of saline solutions. Results: PCR amplification produced the expected fragment sizes for MPSP gene of T. orientalis, msp4 gene of Anaplasma marginale, 16S rRNA gene of C. M. haemobos, RoTaT1.2VSG gene of T. evansi. Nucleotide blast demonstrated that sequences of the PCR amplicons showed a high degree of molecular similarity in comparison with reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16S rRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys as additional species detected in these cattle. Theileria species was the most detected blood pathogen in the sampled cattle. The blood picture of all cattle group revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. Erythrocyte fragility values of all the cattle groups were above the reference range. Anaemia of the macrocytic normochromic type was observed in the Trypanosoma evansi; and Anaplasma platys + Theileria sinensis double species co-infected cattle. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (p<0.05) increases in serum liver and kidney parameters, total protein, globulin, total and unconjugated bilirubin and decreased albumin was observed in the Trypanosoma evansi infected cattle.Conclusion: We present the first report of anaemia associated with Theileria sinensis infection in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of Anaplasma platys, there is an urgent need for appropriate preventive and control measures, as Theileria species and A. platys are of great economic and zoonotic importance respectively.

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Parasitological and molecular detection of Trypanosoma spp. in cattle, goats and sheep in Somalia

Parasitology ◽

10.1017/s003118202000178x ◽

2020 ◽

Vol 147 (14) ◽

pp. 1786-1791

Author(s):

Ahmed A. Hassan-Kadle ◽

Abdalla M. Ibrahim ◽

Hamisi S. Nyingilili ◽

Abdulkarim A. Yusuf ◽

Rafael F. C. Vieira

Keyword(s):

Internal Transcribed Spacer ◽

Molecular Detection ◽

Trypanosoma Evansi ◽

Buffy Coat ◽

Control Measures ◽

Chain Reaction ◽

Blood Samples ◽

Polymerase Chain ◽

Animal Trypanosomiasis ◽

And Control

AbstractAfrican animal trypanosomiasis (AAT) affects the livestock of 12.3 million Somalis and constrains their development and wellbeing. There is missing data on AAT in the country after the civil war of the 1990s. Therefore, this study has aimed to assess the prevalence of Trypanosoma spp. in 614 blood samples from cattle (n = 202), goats (n = 206) and sheep (n = 206) in Afgoye and Jowhar districts, Somalia using parasitological and molecular methods. Twenty-one out of 614 (3.4%; 95% CI: 2.1–5.2%) and 101/614 (16.4%; 95% CI: 13.6–19.6%) ruminants were positive for Trypanosoma spp. by buffy coat technique (BCT) and internal transcribed spacer 1 (ITS1)-polymerase chain reaction (PCR), respectively. Using ITS1-PCR, the highest prevalence was observed in cattle (23.8%; 95% CI: 18.4–30.1%) followed by goats (17.5%; 95% CI: 12.9–23.3%) and sheep (8.3%; 95% CI: 5.1–12.9%). A total of 74/101 (73.3%; 95% CI: 63.5–81.6%) ruminants were shown coinfection with at least two Trypanosome species. The four T. brucei-positive samples have tested negative for T. b. rhodesiense, by the human-serum-resistance-associated-PCR. Trypanosoma evansi, T. godfreyi, T. vivax, T. brucei, T. simiae and T. congolense were the Trypanosoma species found in this study. This is the first study on the molecular detection of Trypanosoma sp. in ruminants in Somalia. Further investigations and control measures are needed to manage Trypanosomiasis spreading in the country. Studies should also focus on the detection of T. b. rhodesiense in the country.

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Species diversity and spatial distribution of CL/VL vectors: assessing bioclimatic effect on expression plasticity of genes possessing vaccine properties isolated from wild-collected sand flies in endemic areas of Iran

BMC Infectious Diseases ◽

10.1186/s12879-021-06129-0 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Ali Bordbar ◽

Parviz Parvizi

Keyword(s):

Gene Expression ◽

Spatial Distribution ◽

Interpolation Method ◽

Diversity Index ◽

Fold Change ◽

Control Measures ◽

Pcr Analysis ◽

Physiological Status ◽

Sand Flies ◽

Risk Maps

Abstract Background Leishmaniasis is one of the ten most important neglected tropical diseases worldwide. Understanding the distribution of vectors of visceral and cutaneous leishmaniasis (VL/CL) is one of the significant strategic frameworks to control leishmaniasis. In this study, the extent of the bioclimatic variability was investigated to recognize a rigorous cartographic of the spatial distribution of VL/CL vectors as risk-maps using ArcGIS modeling system. Moreover, the effect of bioclimatic diversity on the fold change expression of genes possessing vaccine traits (SP15 and LeIF) was evaluated in each bioclimatic region using real-time PCR analysis. Methods The Inverse Distance Weighting interpolation method was used to obtain accurate geography map in closely-related distances. Bioclimatic indices were computed and vectors spatial distribution was analyzed in ArcGIS10.3.1 system. Species biodiversity was calculated based on Shannon diversity index using Rv.3.5.3. Expression fold change of SP15 and LeIF genes was evaluated using cDNA synthesis and RT-qPCR analysis. Results Frequency of Phlebotomus papatasi was predominant in plains areas of Mountainous bioclimate covering the CL hot spots. Mediterranean region was recognized as an important bioclimate harboring prevalent patterns of VL vectors. Semi-arid bioclimate was identified as a major contributing factor to up-regulate salivary-SP15 gene expression (P = 0.0050, P < 0.05). Also, Mediterranean bioclimate had considerable effect on up-regulation of Leishmania-LeIF gene in gravid and semi-gravid P. papatasi population (P = 0.0109, P < 0.05). Conclusions The diversity and spatial distribution of CL/VL vectors associated with bioclimatic regionalization obtained in our research provide epidemiological risk maps and establish more effectively control measures against leishmaniasis. Oscillations in gene expression indicate that each gene has its own features, which are profoundly affected by bioclimatic characteristics and physiological status of sand flies. Given the efficacy of species-specific antigens for vaccine production, it is essential to consider bioclimatic factors that have a fundamental role in affecting the regulatory regions of environmentally responsive loci for genes used in vaccine design.

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Conserved gene sequences for species identification: PCR analysis of the 3′ UTR of the SON gene distinguishes human and other mammalian DNAs

Forensic Science International ◽

10.1016/0379-0738(95)01748-8 ◽

1995 ◽

Vol 73 (3) ◽

pp. 171-181 ◽

Cited By ~ 5

Author(s):

B. Soteriou ◽

R.A. Fisher ◽

I.M. Khan ◽

A.M. Kessling ◽

L.C. Archard ◽

...

Keyword(s):

Species Identification ◽

Pcr Analysis ◽

Gene Sequences ◽

Conserved Gene

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Molecular detection of Anaplasma platys infection in free-roaming dogs and ticks from Kenya and Ivory Coast

Parasites & Vectors ◽

10.1186/s13071-016-1443-3 ◽

2016 ◽

Vol 9 (1) ◽

Cited By ~ 17

Author(s):

Ioana Adriana Matei ◽

Gianluca D’Amico ◽

Patrick K. Yao ◽

Angela Monica Ionică ◽

Paul W. N. Kanyari ◽

...

Keyword(s):

Ivory Coast ◽

Molecular Detection ◽

Anaplasma Platys

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First Molecular Detection of Anaplasma platys and Coinfection with Babesia gibsoni in Dogs from Bengaluru, India

International Journal of Current Microbiology and Applied Sciences ◽

10.20546/ijcmas.2020.903.009 ◽

2020 ◽

Vol 9 (3) ◽

pp. 75-79

Author(s):

Medha Karnik ◽

Anjan Kumar ◽

M. Manjula ◽

H. D. Lohitha ◽

R. Narendra ◽

...

Keyword(s):

Molecular Detection ◽

Anaplasma Platys ◽

Babesia Gibsoni

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Short communication: Molecular detection of honeybee viruses in Ecuador

Spanish Journal of Agricultural Research ◽

10.5424/sjar/2020181-15779 ◽

2020 ◽

Vol 18 (1) ◽

pp. e05SC02

Author(s):

María E. Bravi ◽

Jorge Avalos ◽

Hugo Rosero ◽

Gerald Maldonado ◽

Francisco J. Reynaldi ◽

...

Keyword(s):

Agricultural Production ◽

Molecular Detection ◽

Short Communication ◽

Rna Extraction ◽

Clinical Signs ◽

Pcr Analysis ◽

Pollination Services ◽

Multiple Risk Factors ◽

The World ◽

Honeybee Population

Aim of study: The honeybee, Apis mellifera, is one of the most important pollinators in the world. Apicultural activity and pollination services have been affected by the decline in the honeybee population, which may be due to the interaction of multiple risk factors, such as changes in agricultural production, use of pesticides and presence of pathogens. Viruses, in particular, are suspected to be drivers of colony mortality. In this scenario, the aim of this study was to determine the presence of honeybee viruses (IAPV, DWV, SBV, ABPV, BQCV, CBPV) in A. mellifera populations using a RT-mPCR assay.Area of study: Apiaries were situated in Pichincha, Ecuador.Material and methods: Samples were collected from seventeen apiaries that exhibited mortality but without specific clinical signs. Each sample comprised 15 individuals. After RNA extraction, a multiplex PCR analysis was performed for presence of six viruses (IAPV, DWV, SBV, ABPV, BQCV, CBPV).Main results: Four of the viruses (ABPV, DWV, BQCV and SBV) were found in co-infections in these colonies, with ABPV and SBV also being found in simple infections.Research highlights: To our knowledge, this is the first molecular detection of BQCV and SBV in Ecuador. These findings suggest that some of the above viruses could be involved in weakening these colonies.

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Screening of Highly Expressed Mycobacterial Genes Identifies Rv3615c as a Useful Differential Diagnostic Antigen for the Mycobacterium tuberculosis Complex

Infection and Immunity ◽

10.1128/iai.00150-08 ◽

2008 ◽

Vol 76 (9) ◽

pp. 3932-3939 ◽

Cited By ~ 69

Author(s):

Ben Sidders ◽

Chris Pirson ◽

Philip J. Hogarth ◽

R. Glyn Hewinson ◽

Neil G. Stoker ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Significant Proportion ◽

Growth Conditions ◽

Control Measures ◽

Human Populations ◽

Infected Cattle ◽

Ifn Γ ◽

Mycobacterial Antigens ◽

And Control ◽

Differential Antigens

ABSTRACT Tuberculous infections caused by mycobacteria, especially tuberculosis of humans and cattle, are important both clinically and economically. Human populations can be vaccinated with Mycobacterium bovis bacille Calmette-Guérin (BCG), and control measures for cattle involving vaccination are now being actively considered. However, diagnostic tests based on tuberculin cannot distinguish between genuine infection and vaccination with BCG. Therefore, identification of differential diagnostic antigens capable of making this distinction is required, and until now sequence-based approaches have been predominant. Here we explored the link between antigenicity and mRNA expression level, as well as the possibility that we may be able to detect differential antigens by analyzing quantified global transcriptional profiles. We generated a list of 14 candidate antigens that are highly expressed in Mycobacterium tuberculosis and M. bovis under a variety of growth conditions. These candidates were screened in M. bovis-infected and naïve cattle for the ability to stimulate a gamma interferon (IFN-γ) response. We identified one antigen, Rv3615c, which stimulated IFN-γ responses in a significant proportion of M. bovis-infected cattle (11 of 30 cattle [37%] [P < 0.01]) but not in naïve or BCG-vaccinated animals. Importantly, the same antigen stimulated IFN-γ responses in a significant proportion of infected cattle that did not respond to the well-characterized mycobacterial antigens ESAT-6 and CFP-10. Therefore, use of the Rv3615c epitope in combination with previously described differential tests based on ESAT-6 and CFP-10 has the potential to significantly increase diagnostic sensitivity without reducing specificity in BCG-vaccinated populations.

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