ABSTRACT
The deltamethrin resistance status in Rhipicephalus (Boophilus) annulatus and R. (B.) microplus ticks collected from cattle of five organized farms of Kerala, south India was evaluated. Resistance was characterized using biological (larval packet test), biochemical (esterase enzyme activity assay) and molecular tools (PCR amplification and sequencing of deltamethrin resistance-associated genes). Characterization of field isolates revealed level I resistance in ticks collected from four out of five farms. Elevated level of α/ß esterase activity was not recorded in isolates showing level I resistance. Previously reported point mutations in the carboxyl esterase (G1120A) and sodium channel (T2134A and C190A) genes were not observed in any of the field isolates. The present study showed a low level (level I) resistance is developed in the most economically important ticks infesting cattle of this state and it cautions the development of large scale resistance in future.
Subject(s)
Cattle Diseases/parasitology , Drug Resistance/genetics , Nitriles/pharmacology , Pyrethrins/pharmacology , Rhipicephalus/genetics , Tick Infestations/veterinary , Animals , Arthropod Proteins/genetics , Cattle , Esterases/genetics , Female , India , Male , Point Mutation , Sodium Channels/genetics , Tick Infestations/parasitologyABSTRACT
The objective of the present study is to determine the phylogenetic position of the Theileria organisms in blood of cattle of southern India using molecular tools. Theileria annulata (Namakkal isolate, Tamil Nadu) and three Theileria field isolates (free of T. annulata) from Wayanad, Kerala (Wayanad 1, 2, 3) were used. The small subunit ribosomal RNA (SSU rRNA) and major piroplasm surface protein (MPSP) gene products were cloned, sequenced and the phylogenetic tree constructed. SSU rRNA gene of Wayanad 1 isolate (JQ706077) revealed maximum identity with Theileria velifera or Theileria cervi. The phylogenetic tree constructed based on SSU rRNA genes revealed that Wayanad 1 isolate belonged to a new type which share common ancestor with all the other theilerial species while Wayanad 2 and 3 isolates (JX294459, JX294460) were close to types A and C respectively. Based on MPSP gene sequences, Wayanad 2 and 3 (JQ706078, JX648208) isolates belonged to Type 1 and 3 (Chitose) respectively. When, the previously reported MPSP type 7 is also considered from the same study area, Theileria orientalis types 1, 3 and 7 are observed in south India. SSU rRNA sequence of South Indian T. annulata (JX294461) showed a maximum identity with Asian isolates while the Tams1 merozoite surface antigen (MSA) gene (JX648210) showed maximum identity with north Indian isolate.
Subject(s)
Cattle Diseases/parasitology , Theileria annulata/classification , Theileria annulata/genetics , Theileriasis/parasitology , Animals , Antigens, Protozoan/genetics , Cattle , Cluster Analysis , DNA, Protozoan/chemistry , DNA, Protozoan/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Genes, rRNA , India , Molecular Sequence Data , Phylogeny , Protozoan Proteins/genetics , RNA, Protozoan/genetics , RNA, Ribosomal, 18S/genetics , Sequence Analysis, DNA , Theileria annulata/isolation & purificationABSTRACT
Carriers of bovine anaplasmosis in Northern Kerala, South India were detected using conventional microscopical and molecular techniques. PCR-RFLP and nested PCR techniques were used for detection of Anaplasma marginale and Anaplasma bovis respectively and the PCR products were confirmed by sequencing. Out of 150 samples tested, 25 were detected positive for A. marginale and five for A. bovis based on molecular tests. The inclusion bodies of A. marginale could be detected by microscopy in two blood smears after staining by giemsa while acridine orange staining detected three smears positive. The data clearly suggest the higher sensitivity of molecular techniques for diagnosis of these diseases.
Subject(s)
Anaplasma/isolation & purification , Anaplasmosis/epidemiology , Anaplasmosis/microbiology , Carrier State/veterinary , Cattle Diseases/epidemiology , Cattle Diseases/microbiology , Animals , Bacteriological Techniques , Blood/microbiology , Carrier State/epidemiology , Carrier State/microbiology , Cattle , Cross-Sectional Studies , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , India , Microscopy , Molecular Sequence Data , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Prevalence , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
Carriers of bovine anaplasmosis in Northern Kerala, South India were detected using conventional microscopical and molecular techniques. PCR-RFLP and nested PCR techniques were used for detection of Anaplasma marginale and Anaplasma bovis respectively and the PCR products were confirmed by sequencing. Out of 150 samples tested, 25 were detected positive for A. marginale and five for A. bovis based on molecular tests. The inclusion bodies of A. marginale could be detected by microscopy in two blood smears after staining by giemsa while acridine orange staining detected three smears positive. The data clearly suggest the higher sensitivity of molecular techniques for diagnosis of these diseases.
ABSTRACT
The disease condition attributed to have been caused by Theileria orientalis is generally benign. However, it is also thought that the parasite, at least some strains of it, can cause fatal disease. The present communication deals with the clinical signs, postmortem lesions and diagnosis of a fatal disease due to T. orientalis which caused mortality in crossbred adult bovines of South India. High body temperature, lacrimation, nasal discharge, swollen lymph nodes and haemoglobinuria were the symptoms observed. The postmortem lesions observed were punched out ulcers in abomasum, enlargement of spleen, massive pulmonary oedema, frothy exudates in trachea, epicardial and endocardial haemorrhage and haemorrhagic duodenitis. Peripheral blood smear examination revealed rod shaped Theileria sp. organisms. Polymerase chain reaction that amplify the T. orientalis specific P(32/33) gene, followed by cloning and sequencing, revealed maximum homology with Narathiwat (Thailand) and Jingole -1 (Indonesia) isolates which were positioned as isolate type 7 of T. orientalis.
