First Survey on the Presence and Distribution of Oxytetracycline-Resistance Genes in Anaplasma Species.

INTRODUCTION: Anaplasma sp. is an obligatory intracellular Gram-negative tick-transmitted bacterial pathogen of humans and animals. Oxytetracycline and chlortetracycline are the drugs of choice for treating domestic animals with acute anaplasmosis. Lack of documented information about oxytetracycline resistance in Anaplasma species in the world was the scope of this study to screen by PCR for the detection of the oxytetracycline-resistance genes in Anaplasma species from infected cattle and sheep in the Northwest and Southwest of Iran. MATERIALS AND METHODS: Total of 100 cattle and sheep blood samples collected from 2 provinces in the Northwest and 1 province in the Southwest of Iran were tested microscopically by the Giemsa staining examination and confirmed by PCR. Then the presence of two different oxytetracycline-resistance genes (otrA, and otrB) was detected by PCR in positive samples. RESULTS: The results showed that 60% of Anaplasma-infected samples were identified to have an otrA-resistance gene, and 26.67% had an otrB-resistance gene. The coexistence of two oxytetracycline-resistance determinants was encountered in 13.33% of the isolates. The significant difference in the frequency of otr genes was found among three Anaplasma species (A. marginale, A. centrale and A. ovis), and among three studied regions in Iran (p < 0.05). The identified sequences were submitted to the GenBank and deposited under accession numbers MN880729 and MN895439 for otrB and otrA genes. CONCLUSION: This study, for the first time, indicated the oxytetracycline-resistance genes in the three most prevalent Anaplasma species in ruminants. This finding helps to select an appropriate treatment strategy for eradication of anaplasmosis.

Molecular study and genotyping of Cryptosporidium baileyi and Cryptosporidium parvum from free-range and commercial broiler chickens in Guilan province, Iran.

Cryptosporidiosis acutely impacts the digestive and/or respiratory tract of the birds in many species of various orders. More importantly, it is also well known as a significant zoonotic disease, which can lead to diarrhea in humans and livestock. Regarding increasing demand for free-range products and increasing the number of free-range poultry farms, the present paper evaluated histopathological and molecular detection of Cryptosporidium baileyi and Cryptosporidium parvum in free-range and commercial broiler chickens in the north part of Iran. For this purpose, 100 fecal and tissue samples of the chickens in Guilan province were collected. After microscopic examination using Ziehl-Neelsen staining, molecular analyses of the fecal samples were processed by Nested-PCR targeting the 18S rRNA gene followed by sequencing of the amplicons and phylogenetic analyses. Eventually, the tissue samples were studied for histological lesions. Findings demonstrated the presence of Cryptosporidium baileyi and Cryptosporidium parvum in 6 % and 2 % of fecal samples, respectively. This is the first identification of C.parvum in avian hosts in Iran, and for the first time, C.baileyi and C.parvum are shown in native free-range chickens in Iran. All of the PCR positive birds with clinical symptoms showed gross lesions of respiratory infections. There was no significant difference between infection rate in free-range and commercial broiler chickens; however, the infection rate was significantly higher in chickens <25 days old. To conclude, we present here a notable Cryptosporidium infection rate in the free-range chicks in Iran, which notify the role of this host as a reservoir and should be more noted due to the economic and zoonotic importance.