The study investigated the occurrence of trypanosomosis in water buffaloes using parasitological and molecular methods. Buffaloes of different ages and both sexes comprising Bulgarian Murrah and some crosses were used in the study. Two of the 145 samples (0.13%) were positive for Trypanosoma evansi in both, blood parasite examination and PCR. One female buffalo (1/54 samples; 1.86%) from a lactating herd in the city of Muñoz, Nueva Ecija (Region 3) and another (1/51 samples; 1.96%) non-pregnant female belonging to caraheifer population from Los Baños, Laguna (Region 4) were found infected by T. evansi. The positive animals were in the age group of 3-4 years. Blood samples obtained from 40 buffaloes from Cagayan (Region 2) were found negative for trypanosome infection.
Abstract:Hepatozoon canis infection in canines is allegedly an underreported disease in the Philippines. In over the past four decades, there are only two published cases in the country. A total of 168 canine blood samples was processed for polymerase chain reaction (PCR) amplification and blood parasite examination (BPE). The PCR method was able to provide molecular evidence for the presence of Hepatozoon canis genomic DNA in one sample (0.6%). Consequently, Hepatozoon canis gametocytes demonstrating the classical elongated appearance in leucocytes were also consistently seen on Giemsa ® -stained blood smears of the PCR-positive animal after BPE. The study elucidates the parasitological detection and first molecular evidence of Hepatozoon canis infection in the Philippines by PCR.
Background: Trypanosoma brucei rhodesiense is a haemoflagellate parasite of zoonotic significance. Aside from its public health importance, this parasite subspecies gained notoriety because of their effective system to circumvent the immune response of vertebrate host. The parasite cell surface is covered with millions of VSG dimers, which serve as an almost infinite repertoire of biomolecules needed for evasion of host immune system. Around two decades ago, it was resolved that all trypanosome VSG is associated with one or more N-linked oligosaccharides, with a range of structures including high mannose and complex types. This complex process of protein modification known as N-linked glycosylation is catalyzed by oligosaccharyl transferase (OST). In general, the incorporation of glycan structures can alter protein's antigenic properties and recently it was established that glycan molecules associated with VSG were found to be important in several aspects of trypanosome-host interaction, especially during parasite evasion of the host defense mechanisms. Therefore, our major interest is to clone and characterize the trypanosome OST. Material, Methods and Results: The template genomic DNA for PCR amplification was extracted as described previously. In an attempt to clone Trypanosoma brucei rhodesiense putative oligosaccharyl transferase, an amplicon of ~2000 bp was obtained having an open reading frame of 2057 bp and deduced primary structure composed of 685 amino acid residues (TbrOST II). Comparison of TbrOST II ORF with annotated putative oligosaccharyl transferase in the genome of other organisms revealed sequence identity to other kinetoplastid. TbrOST II had high nucleotide (Ns) and amino acid (As) sequence similarity with the genomes of T. brucei gambiense (Ns:99%; As:78%) and T. brucei (Ns:95-98%; As:77%-98%). There was also significant nucleotide and amino acid sequence identity in the genomes of T. cruzi (Ns:74%; As:63%), Leishmania infantum (Ns:70-83%; As:46-57%), L. braziliensis (Ns:69-81%; As:46-55%) and L. major (Ns:69-80%; As:46-57%). Sequence similarity (71-77%) from other origins was also exhibited. The nucleotide sequence alignments and analysis were performed using the Oxford University Mac Vector 6.5 sequence analysis software and CLC Workbench 5.6 software. Discussion: The nucleotide BLAST results indicate that sequence identity is higher between species of the same genus rather than of the same family. It is known that T. brucei, T. gambiense and T. rhodesiense are members of the Brucei-complex or Brucei group. Although T. brucei brucei has more similarities with T. brucei rhodesiense than T. brucei gambiense, these parasites are morphologically indistinguishable. This is the probable reason why high sequence identity was displayed by other subspecies of the Brucei group. In addition, the high percent identity possessed by TbrOST II with other trypansomatids agrees with the evolutionarily conserved characteristics of the established OST. The DNA sequence data of TbrOST II showing similar seque...
Abstract:Trypanosoma theileri is one of the protozoan parasites reported in cattle and carabaos in the Philippines. The distribution of T. theileri infection in livestock in this country is not well established. To date, more than two decades have passed without any new T. theileri prevalence reported in the country. The present study endeavored to determine whether the parasite is endogenously present in cattle from the remote areas of Region II (Cagayan Valley), particularly in the province of Quirino, Philippines and to establish the current parasite prevalence of T. theileri in this province using the blood parasite examination method. A total of 246 field blood samples of cattle was collected from 5/6 (83%) of the municipalities of Quirino Province, Philippines. Blood parasite examination of Giemsa-stained smears revealed that all samples were negative for the presence of the T. theileri parasite. Given that the mainstay of parasitological examination in the field is the classic blood parasite examination method, the data suggest that the negative results of these examinations can probably be attributed to very low parasitemia.
The partial nucleotide sequence of putative Trypanosoma brucei rhodesiense oligosaccharyl transferase gene was previously reported. Here, we describe the determination of its full-length nucleotide sequence by Inverse PCR (IPCR), subsequent biological sequence analysis and transmembrane topology modelling. The full-length DNA sequence has an Open Reading Frame (ORF) of 2406 bp and encodes a polypeptide of 801 amino acid residues. Protein and DNA sequence analyses revealed that homologues within the genome of other kinetoplastid and various origins exist. Protein topology analysis predicted that Trypanosoma brucei rhodesiense putative oligosaccharyl transferase clone II (TbOST II) is a transmembrane protein with transmembrane helices in probably an N(cytosol)-C(cytosol) orientation. Data from the GenBank database assembly and sequence analyses in general clearly state that TbOST II is the STT3 subunit of OST in T.b. rhodesiense that necessitates further characterisation and functional studies with RNAi. TbOST II sequence had been deposited in the GenBank (accession number GU245937).
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