First Molecular Survey on Anaplasma phagocytophilum Re-vealed High Prevalence in Rural Dogs from Khuzestan Prov-ince, Iran

Background: Anaplasmosis due to Anaplasma phagocytophilum is an important tick-borne zoonotic disease, which affects dogs, horses, cattle and human as well. This study aimed to probe the existence of this organism by means of molecular biology techniques for the first time in rural dogs of Khuzestan province, Southwestern Iran. Methods: During Sep 2014 to Apr 2015 blood samples of 103 apparently healthy rural dogs (60 males) were collected for A. phagocytophilum detection by light microscopical examination of Giemsa stained slides and Nested PCR on a fragment of 16S rRNA gene. Results: From the examined slides, 11.65% were positive for A. morulae while 57.28% of infection was revealed by Nested PCR method. There was no statistical difference between ages and sexes of dogs and infection in molecular survey of A. phagocytophilum. Conclusion: Molecular prevalence of A. phagocytophilum was noticeably high. It may cause the incidence of disease in human population.

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Evaluation of Mycoplasma arginini abundance in slaughtered camels with pneumonia confirm with nested PCR method for the first time in Iran

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2011.08.757 ◽

2011 ◽

Vol 44 (13) ◽

pp. S300

Author(s):

Izadi Maryam ◽

Navid Mehr Jafar ◽

Lal Alizadeh Samane ◽

Bolori Mehrin ◽

Bahmani Somaye

Keyword(s):

Nested Pcr ◽

Pcr Method ◽

First Time

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KARAKTERISASI FITOPLASMA PENYEBAB PENYAKIT LAYU KELAPA DI PULAU DERAWAN MENGGUNAKAN RFLP IN SILICO

JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA ◽

10.23960/j.hptt.217105-110 ◽

2018 ◽

Vol 17 (2) ◽

pp. 105

Author(s):

Agus Eko Prasetyo ◽

Kikin Hamzah Mutaqin ◽

Giyanto .

Keyword(s):

Nested Pcr ◽

In Silico ◽

Bacterial Species ◽

Rflp Analysis ◽

Wilt Disease ◽

Rrna Gene ◽

16S Rrna Gene Sequences ◽

Gram Positive Bacteria ◽

Pcr Method

Characterization of phytoplasmas associated with coconut wilt disease in Derawan Island using in silico RFLP. Coconutwilt disease has been reported in Derawan Island that resulted in eradication up to 10% of the total cultivated palms. Theobjective of this study was to detect and characterize phytoplasmas associated with coconut wilt disease in Derawan islandusing nested PCR technique and in silico RFLP based on 16S rRNA gene sequences. Detection of phytoplasmas was performedusing nested PCR method, cloning of nPCR products, sequencing, and analysis of sequencing results using in silico RFLP.The results revealed that phytoplasmas could not be detected by PCR using P1/P7 primer pairs however it could be amplifiedby nested PCR using R16F2n/R16R2 primer pairs resulting amplicon at about 1.25 kb. In silico RFLP analysis indicated thatphytoplasmas associated with coconut wilt disease in Derawan Island belong to 16SrII (witches broom phytoplasma). PCRproduct of the nPCR need to be sequenced because the R16F2n/R16R2 primer will also amplify the other bacterial species, mainly from Gram positive bacteria.

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Study of the genus Bracon Fabricius, 1804 (Hymenoptera: Braconidae) from Iran: new subspecies, new records and an updated checklist

Zootaxa ◽

10.11646/zootaxa.4758.2.1 ◽

2020 ◽

Vol 4758 (2) ◽

pp. 201-230

Author(s):

MOHAMMAD ZARGAR ◽

KONSTANTIN SAMARTSEV ◽

ALI ASGHAR TALEBI ◽

SAMIRA FARAHANI

Keyword(s):

New Records ◽

Distribution Data ◽

Southwestern Part ◽

New Subspecies ◽

Khuzestan Province ◽

Southwestern Iran ◽

First Time

The present study is based on the genus Bracon Fabricius collected from the Khuzestan province in the southwestern part of Iran during 2016–2017. In total, 35 species including one subspecies were collected and identified, of which 10 species are recorded for the first time from Iran and Bracon (Osculobracon) pelliger rumezensis Samartsev & Zargar ssp. n. is described. In addition, 24 species are recorded for the first time from Khuzestan province in southwestern Iran. Brief diagnoses with illustrations for new records from Iran and a faunistic list with distribution data are provided.

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Detection and Identification of Banana-associated Phytoplasma Using Nested-PCR Method

Jurnal Perlindungan Tanaman Indonesia ◽

10.22146/jpti.38320 ◽

2019 ◽

Vol 23 (1) ◽

pp. 148

Author(s):

Saurma Mona Astrid Sibarani ◽

Tri Joko ◽

Siti Subandiyah

Keyword(s):

16S Rrna ◽

Nested Pcr ◽

Plant Diseases ◽

16S Rrna Genes ◽

Rrna Genes ◽

Rrna Gene ◽

Aster Yellows ◽

Detection And Identification ◽

Pcr Method ◽

The 16S Rrna Gene

Phytoplasma is known to be associated with plant diseases in about 300 plant species from various families. Information on the presence of phytoplasma in bananas as one of the pathogens that can cause disease in bananas in Indonesia has never been reported. This research was conducted with the aim to detect the presence of banana phytoplasma by the nested-PCR method and to identify phytoplasma obtained based on the sequence analysis of the 16S rRNA gene. Standard PCR was carried out using P1/P7 primary pairs, followed by nested-PCR using a pair of R16F2n/R16R2m23SR primers separately that could amplify the target 16S rRNA genes in a row at 1600 bp. BLAST analysis shows that the results of phylogenetic analysis of banana phytoplasmic nucleotide cv. manggala from Tasikmalaya and cv. Raja nangka from Banjar has a genetic relationship that is closer to lethal wilt oil palm Phytoplasma (Candidatus Phytoplasma asteris). This phytoplasma belongs to the 16SrI-B group (aster yellows).

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The genus Basiria Siddiqi, 1959 (Nematoda: Tylenchidae) from Dezful region, Iran

Journal of Threatened Taxa ◽

10.11609/jott.6353.13.3.18004-18010 ◽

2021 ◽

Vol 13 (3) ◽

pp. 18004-18010

Author(s):

Manouchehr Hosseinvand ◽

Ali Eskandari ◽

Reza Ghaderi

Keyword(s):

Body Length ◽

Cephalic Region ◽

Parasitic Nematodes ◽

Plant Parasitic Nematodes ◽

Morphometric Characters ◽

Khuzestan Province ◽

Basal Bulb ◽

Southwestern Iran ◽

First Time ◽

Plant Parasitic

A survey was conducted during 2018 and 2019 in order to identify plant-parasitic nematodes of the genus Basiria in Dezful region of Khuzestan province, southwestern Iran. Nematodes were extracted from the soil and root samples by using tray method, transferred to glycerin and mounted on permanent slides. Nematodes were identified based on morphological and morphometric characters. As a result, eight species including B. aberrans, B. duplexa, B. gracilis, B. jirians, B. tumida, B. graminophila, B. ritteri, and B. similis were identified; three species namely B. jirians, B. ritteri, and B. similis are here described and illustrated for the first time from Iran. B. jirians is characterized by body length 445–535 µm, stylet 9.0–9.2 µm, cephalic region without annuli, DGO 2.0–2.5 µm, median bulb at anterior end of pharynx, basal bulb pyriform, spermatheca non offset and tail elongate conoid with pointed to filiform terminus. B. ritteri can be characterized by body length 685–747 µm, stylet 10.5–11.5 µm, median bulb located at anterior half of pharynx, basal bulb cylindroid, spermatheca non offset and tail annulated and notched at tip. B. similis is characterized by body length 644–736 µm, stylet 10.3–11 µm, DGO 8.9–10.5 µm, basal bulb cylindroid and tail clavate.

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Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand

BMC Veterinary Research ◽

10.1186/s12917-020-02585-z ◽

2020 ◽

Vol 16 (1) ◽

Author(s):

Anh H. L. Nguyen ◽

Sonthaya Tiawsirisup ◽

Morakot Kaewthamasorn

Keyword(s):

Water Buffalo ◽

Pcr Amplification ◽

Reservoir Host ◽

Rrna Gene ◽

Homology Searching ◽

The Family ◽

High Prevalence ◽

Control And Prevention ◽

Percentage Similarity ◽

First Time

Abstract Background Anaplasmosis, an animal disease caused by rickettsial bacteria in the genus Anaplasma, is of considerable economic importance in livestock animals in many countries worldwide. The objectives of this study were to determine the identity, prevalence, and geographic distribution of Ehrlichia and Anaplasma in naturally infected water buffalo in Thailand using PCR amplification and sequencing of the 16S ribosomal RNA and heat shock protein groEL genes. A total of 456 buffalo blood samples from Thailand were investigated. Species identification and genetic differentiation of intra-population and inter-population with the global isolates were conducted based on nucleotide sequences. Interplay between the infection and host factors was also assessed. Results Overall, 41% of water buffalo were found to be infected with rickettsial organisms in the family Anaplasmataceae, but Ehrlichia spp., Neorickettsia spp., and Wolbachia spp. were not found in any of the sequenced samples in this study. Female buffalo were more frequently infected with bacteria in the family Anaplasmataceae than males [71 out of 176 females (40.3%) versus 11 out of 47 males (23.4%)]. The Odds Ratio value indicated that the risk of infection for female buffalo was 2.2-fold higher than that for males (p < 0.05). We detected three haplotypes of A. marginale 16S rRNA gene and they were placed in a clade that was closely related to the A. marginale in buffalo in China; and cattle in Thailand, Uganda, and China. Homology searching of groEL sequences against the GenBank™ database using the BLASTn algorithm revealed that the obtained sequences had a high percentage similarity (98.36–99.62%) to A. platys sequences. The groEL sequences of three A. platys-like isolates were clustered in the same clade as the A. platys from the tick Rhipicephalus microplus in China. Conclusions Our data showed that the apparently healthy buffalo were naturally infected by bacteria in the family Anaplasmataceae at a relatively high prevalence. We also report the finding of A. platys-like infections in water buffalo in Thailand for the first time. Water buffalo serving as the reservoir host of anaplasmosis is of concern for managing the disease control and prevention in ruminants.

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Prevalent latent human cytomegalovirus genotype b2 in biopsy samples of gastric cancer

Future Virology ◽

10.2217/fvl-2019-0117 ◽

2020 ◽

Vol 15 (2) ◽

pp. 71-78

Author(s):

Alireza Mohebbi ◽

Zoleikha Mamizadeh ◽

Hanieh Bagheri ◽

Farzad Sharifnezhad ◽

Alijan Tabarraei ◽

...

Keyword(s):

Gastric Cancer ◽

Viral Load ◽

Gastrointestinal Tract ◽

Human Cytomegalovirus ◽

Nested Pcr ◽

Viral Loads ◽

Pcr Method ◽

H Pylori ◽

Viral Inclusions ◽

First Time

Aim: Genotyping and prevalence of human cytomegalovirus (HCMV) in gastrointestinal tract remains unclear. Objective: Characterization of HCMV and its viral load in samples of patients with gastric cancer (GC) in northeast Iran. Methods: A total of 21 biopsies were collected from patients with GC. HCMV was detected and genotyped using multiplex nested PCR. Quantitative real-time PCR has been used to determine the viral load. Staining was performed to detect viral inclusions. Results: 76.19% of the samples had HCMV genotypic b2 with viral load <0.1 IU/μl. 66.67% of patients were positive for H. pylori. Conclusion: Multiplex nested-PCR method can detect CMV populations with low viral loads in GC biopsies. For the first time, it was determined that the genotype b2 of HCMV is prevalent in the gastrointestinal tract.

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Concurrent occurrence of Anaplasma phagocytophilum and A. marginale in bovine peripheral blood samples from southwest of Iran

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.25076 ◽

2020 ◽

Vol 71 (3) ◽

pp. 2301

Author(s):

S. BAHRAMI ◽

H. HAMIDINEJAT ◽

MR HAJI HAJIKOLAEI ◽

S. KAVIANIFAR

Keyword(s):

Nested Pcr ◽

Anaplasma Phagocytophilum ◽

Epidemiological Studies ◽

Infected Cattle ◽

Blood Samples ◽

Khuzestan Province ◽

Pcr Product ◽

Health Significance ◽

Cattle Herds ◽

Southwest Of Iran

Anaplasma phagocytophilum and A. marginale are the most important tick-borne bacteria of veterinary and public health significance. Anaplasma phagocytophilum causes febrile disease in humans (human granulocytic-HGA) and animals. Anaplasma marginale is the aetiological agent of acute anaplasmosis, a bovine syndrome characterized by progressive hemolytic anemia associated with fever, weight loss, abortion, decreased milk production, and in some cases, death of the infected cattle. The present study was designed to investigate the prevalence of A. phagocytophilum and A. marginale in cattle from Khuzestan province, southwest of Iran. Samples were collected between March to August 2016. Farmed cattle were selected from the four geographic regions of Khuzestan province with the highest population of cattle herds: Behbahan; Dezful; Shushtar; and Ahvaz. Blood samples were collected from the jugular vein of 200 cattle Species specification was accomplished by specific Nested PCR according to amplification of the 16SrRNA gene. To identify A. marginale, semi-nested PCR product was cut with restriction endonucleases Bst 1107 I. The prevalence of the A. marginale infection (21.5 %) was higher than that of A. phagocytophilum (7.5 %), which was found in a mixed infection with A. marginale. Overall, in the present study 7.5% of cattle were infected with both A. phagocytophilum and A. marginale. Despite the healthy appearance of infected cattle, they can transmit Anaplasma to ticks and are potential continuous sources for maintaining and disseminating the organisms to the human and animals’ population. More epidemiological studies are needed to determine the vectors and reservoir animals for the Anaplasma species and to clarify the pathogenicity of A. marginale and A. phagocytophilum for humans and animals in Iran.

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Detection of Anaplasma phagocytophilum in Wild and Farmed Cervids in Poland

Pathogens ◽

10.3390/pathogens10091190 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1190

Author(s):

Anna W. Myczka ◽

Żaneta Steiner-Bogdaszewska ◽

Katarzyna Filip-Hutsch ◽

Grzegorz Oloś ◽

Michał Czopowicz ◽

...

Keyword(s):

Nested Pcr ◽

Anaplasma Phagocytophilum ◽

Biological Material ◽

Red Deer ◽

Roe Deer ◽

Fallow Deer ◽

Tissue Samples ◽

Natural Reservoir ◽

First Time

Background: The role of cervids in the circulation of A. phagocytophilum has not yet been clearly determined; however, several species of wild and farm cervids may be a natural reservoir of this bacteria. Methods: Spleen and liver tissue samples were taken from 207 wild (red deer, roe deer, fallow deer and moose) and farmed cervids (red deer and fallow deer) from five geographical areas. These were tested for the A. phagocytophilum16S rDNA partial gene by nested PCR. Results: Anaplasma spp. were detected in 91 of 207 examined cervids (prevalence 43.9%). Three different variants of 16S rDNA partial gene were reported, one for the first time. Anaplasma phagocytophilum was more often detected in young specimens than in adults and more often in the spleen than in the liver. Conclusions: Cervids from the four sites across Poland were found to be major natural reservoirs of various strains of A. phagocytophilum. This is the first study to use spleen and liver as biological material to detect A. phagocytophilum in moose in Poland.

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Prevalence of Toxoplasma gondii and Toxocara canis among Myositis Patients in Southwest of Iran

Infectious Disorders - Drug Targets ◽

10.2174/1871526520666191231123159 ◽

2019 ◽

Vol 20 ◽

Author(s):

Jasem Saki ◽

Karim Mowla ◽

Reza Arjmand ◽

Forough Kazemi ◽

Somayeh Fallahizadeh

Keyword(s):

Toxoplasma Gondii ◽

Nested Pcr ◽

Healthy Subjects ◽

Toxocara Canis ◽

Control Group ◽

Igg Antibodies ◽

Healthy Individuals ◽

High Prevalence ◽

Acute Toxoplasmosis ◽

Southwest Of Iran

Introduction: Parasitic myositis is caused by some parasites such as T. gondii and T. canis. So, the aim of the study was to evaluate the prevalence T. gondii and T. canis in patients with myositis and healthy individuals. Methods: A total of 108 samples were randomly selected as the control (54 healthy individuals) and test (54 myositis patients) groups. IgG and IgM antibodies against T. gondii and IgG antibodies against T. canis were measured by the ELISA. The detection of chronic and acute toxoplasmosis was performed by the ELISA IgG avidity. The presence of T. gondii in blood was evaluated by the nested-PCR. Results: Of 108, 33 (30.6%) cases were detected positive for IgG against T. gondii that 19 (35.2%) and 14 (25.9%) were observed in myositis patients and healthy individuals, respectively (P=0.296). Of 19 positive cases, 12 (63.2%) and 7 (36.8%) cases were detected as chronic and acute toxoplasmosis, respectively, while, all positive cases in the control group had chronic toxoplasmosis (P=0.013). One (1.9%) sample was detected positive for anti- Toxoplasma gondii IgM and two (3.7%) samples were found positive for IgG against T. canis by the ELISA that these positive cases were observed only in myositis patients (P=1.000 P=0.495, respectively). B1 T. gondii gene was amplified in 12 (63.2%) and 1 (7.1%) in myositis patients and healthy subjects (P=0.001). Conclusions: Our findings showed that there was a relatively high prevalence of acute toxoplasmosis in myositis patients in comparison with the control subjects in southwest of Iran.

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