Molecular Detection of Novel Genetic Variants Associated to Anaplasma ovis among Dromedary Camels in Iran.

To the best of our knowledge, little information is available regarding the presence of Anaplasma species in camels in Iran. This study sought to investigate the presence of Anaplasma species by microscopy and polymerase chain reaction (PCR) assays in 100 healthy dromedaries (Camelus dromedarius) arriving for slaughter. The microscopic examination of Giemsa-stained blood films revealed that Anaplasma like structures could be identified in the erythrocytes of two blood smears. To confirm the presence of and to identify the species of Anaplasma spp., a PCR technique was performed using primers amplifying a 750 bp fragment of the 16S rRNA gene of Anaplasma and the PCR products were analyzed by sequencing. The nucleotide sequence was compared to the sequences available in GenBank using the Basic Local Alignment Search Tool (BLAST). According to the results, the sequences of two 16S rRNA PCR products clearly fit within the Anaplasma genus in the family Anaplas mataceae. In this study, phylogenetic analysis using the 16S rRNA gene sequences revealed that two sequences obtained from monophyletic clusters included Anaplasma ovis (A. ovis). The obtained sequences had 99.6-100% similarity with previously published 16S rRNA gene sequences. This study aimed to evaluate the presence of novel genetic variants associated to A. ovis in dromedaries in the world. Further studies are recommended to establish the vector(s), as well as the veterinary and medical significance of these apparently novel variants in Iran.To the best of our knowledge, little information is available regarding the presence of Anaplasma species in camels in Iran. This study sought to investigate the presence of Anaplasma species by microscopy and polymerase chain reaction (PCR) assays in 100 healthy dromedaries (Camelus dromedarius) arriving for slaughter. The microscopic examination of Giemsa-stained blood films revealed that Anaplasma like structures could be identified in the erythrocytes of two blood smears. To confirm the presence of and to identify the species of Anaplasma spp., a PCR technique was performed using primers amplifying a 750 bp fragment of the 16S rRNA gene of Anaplasma and the PCR products were analyzed by sequencing. The nucleotide sequence was compared to the sequences available in GenBank using the Basic Local Alignment Search Tool (BLAST). According to the results, the sequences of two 16S rRNA PCR products clearly fit within the Anaplasma genus in the family Anaplas mataceae. In this study, phylogenetic analysis using the 16S rRNA gene sequences revealed that two sequences obtained from monophyletic clusters included Anaplasma ovis (A. ovis). The obtained sequences had 99.6-100% similarity with previously published 16S rRNA gene sequences. This study aimed to evaluate the presence of novel genetic variants associated to A. ovis in dromedaries in the world. Further studies are recommended to establish the vector(s), as well as the veterinary and medical significance of these apparently novel variants in Iran.

The First Record of Argulus foliacesus (Crustacea: Branchiura) Infestation on Lionhead Goldfish (Carassius auratus) in Iran.

Argulus foliaceus (Crustacea: Branchiura), or the fish louse, is an ectoparasite of the skin or gill of the fresh water fish species. Clinical signs in infected fish include scratching on aquarium walls, erratic swimming, and poor growth. It causes pathological changes due to direct tissue damage and secondary infections. In the present study, lionhead goldfish (Carassius auratus), taken from a goldfish aquarium with symptoms such as abnormal swimming, poor growth and death, were examined for ectoparasites. The parasites collected from the skin and fins of fish were identified as A. foliaceus. Then, treatment was carried out by trichlorfon. After administration, no parasite was observed on the fish. This is the first report of infection with A. foliaceus of lionhead goldfish (Carassius auratus) in Iran.

Comparison of Microscopy and PCR-RFLP for detection of Anaplasma marginale in carrier cattle.

BACKGROUND AND OBJECTIVES: In Iran, anaplasmosis is normally diagnosed with traditional Giemsa staining method. This is not applicable for identification of the carrier animals. The aim of this study was to compare the detection of Anaplasma marginale in two different numbers of microscopic fields (50 and 100) using conventional Giemsa staining method compared with the PCR-RFLP technique. MATERIALS AND METHODS: In this study, examinations were performed on 150 blood samples from cattle without clinical signs. Sensitivity and specificity of two microscopic fields (50 and 100 fields) were compared with A. marginale specific PCR-RFLP. The degree of agreement between PCR-RFLP and the two microscopic tests was determined by Kappa (κ) values with 95% confidence intervals. RESULTS: PCR-RFLP showed that 58 samples were A. marginale, while routine microscopy showed erythrocytes harboring Anaplasma like structures in 16 and 75 blood samples determined in 50 and 100 microscopic fields respectively. Examination of 50 and 100 microscopic fields showed 25.8% and 91.4% sensitivity and 99% and 76.1% specificity compared to 100% sensitivity and specificity by PCR-RFLP. The Kappa coefficient between PCR-RFLP and Microscopy (50 fields) indicated a fair level of agreement (0.29). The Kappa coefficient between PCR-RFLP and Microscopy (100 fields) indicated a good level of agreement (0.64) CONCLUSION: Our results showed that the microscopic examination remains the convenient technique for day-to-day diagnosis of clinical cases in the laboratory but for the detection of carrier animal with low bacteremia, microscopy with 100 fields is preferable to Microscopy with 50 fields and molecular methods such as PCR-RFLP can be used as a safe method for identifying cattle persistently infected with A. marginale.