scholarly journals Eimeria stiedae: Infection rate and molecular characterization by nested PCR in rabbits from Minoufiya Governorate, Egypt

2020 ◽  
Vol 16 (1) ◽  
pp. 34-49
Author(s):  
Mahmoud AbouLaila
Keyword(s):  
Molecular Characterization ◽  
Infection Rate ◽  
Nested Pcr ◽  
Eimeria Stiedae

scholarly journals Toxoplasma gondii in Sheep and Goats from Central Iran

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Mojtaba Bahreh ◽  
Bahador Hajimohammadi ◽  
Gilda Eslami
Keyword(s):  
Toxoplasma Gondii ◽  
Dna Extraction ◽  
Infection Rate ◽  
Nested Pcr ◽  
Prevention Programs ◽  
Central Iran ◽  
Infection Rates ◽  
Sheep And Goats ◽  
One Year ◽  
Undercooked Meat

Abstract Objective Toxoplasmosis, caused by Toxoplasma gondii, infects humans by consuming infected raw or undercooked meat and foods harboring mature oocysts. In this study, we assessed the prevalence of T. gondii in sheep and goats coming from central Iran. After completing the questionnaire, about one gram of liver or diaphragm tissue was taken as a sample from 90 sheep and 90 goats slaughtered in Yazd Province and stored at – 20 ºC. DNA extraction was done, and then T. gondii was detected using nested PCR. Results This study indicated that the prevalence of T. gondii in all slaughtered animals was 11.6% (21 of 180), including 14.4% (13/90) in sheep and 8.8% (8/90) in goats. The infection rates in liver and diaphragm samples were 12.2% (11/90) and 11.1% (10/90), respectively (p = 0.8163). The infection rate in animals older than one was 16.3% (15/92), and it was 6.8% (6/88) in animals under one year of age. Therefore, no significant differences were found (p = 0.475). Infection rates were 19.5% (18/92) in males and 3.4% (3/88) in females (p = 0.0007). In conclusion, the infection rates of toxoplasmosis in livestock in this area are almost high, and therefore, it is necessary to design appropriate prevention programs to control the disease.

scholarly journals Molecular Characterization ofCryptosporidiumspp. in Wild Rodents of Southwestern Iran Using 18s rRNA Gene Nested-PCR-RFLP and Sequencing Techniques

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Jasem Saki ◽  
Masoud Foroutan-Rad ◽  
Reza Asadpouri
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
18S Rrna ◽  
Restriction Enzymes ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Wild Rodents ◽  
Nested Polymerase Chain Reaction ◽  
Pcr Rflp ◽  
Southwest Of Iran

Background. Rodents could act as reservoir forCryptosporidiumspp. speciallyC. parvum, a zoonotic agent responsible for human infections. Since there is no information aboutCryptosporidiuminfection in rodents of Ahvaz city, southwest of Iran, hence, this survey was performed to determine the prevalence and molecular characterization ofCryptosporidiumspp. in this region.Materials and Methods. One hundred rodents were trapped from different regions of Ahvaz city. Intestine contents and fecal specimens of rodents were studied using both microscopy examination to identify oocyst and nested-polymerase chain reaction (PCR) technique for 18s rRNA gene detection. Eventually restriction fragment length polymorphism (RFLP) method usingSspIandVspIrestriction enzymes was carried out to genotype the species and then obtained results were sequenced.Results. Three out of 100 samples were diagnosed as positive and overall prevalence ofCryptosporidiumspp. was 3% using both modified Ziehl-Neelsen staining under light microscope and nested-PCR (830 bp) methods. Afterwards, PCR-RFLP was performed on positive samples andC. parvumpattern was identified. Finally PCR-RFLP findings were sequenced and presence ofC. parvumwas confirmed again.Conclusions. Our study showed rodents could be potential reservoir forC. parvum. So an integrated program for control and combat with them should be adopted and continued.

Molecular characterization of Candidatus Phytoplasma aurantifolia isolate infecting chickpea (Cicer arietinum) in Dharwad, Karnataka

2019 ◽  
Author(s):  
Gurupada Balol ◽  
C Channakeshava ◽  
M S Patil
Keyword(s):  
16S Rdna ◽  
Molecular Characterization ◽  
Cicer Arietinum ◽  
Nested Pcr ◽  
Rdna Sequences ◽  
Pcr Product ◽  
16S Rdna Sequences ◽  
Axillary Proliferation ◽  
Pale Green

Chickpea plants showing phytoplasma symptoms were observed in the research plots at University of Agricultural Sciences, Dharwad, Karnataka, India. The symptoms included phyllody, pale green leaves, bushy appearance and excessive axillary proliferation. The causal agent of the phyllody disease was identified based on symptoms, amplification of 16S rDNA of the phytoplasma by nested PCR with primers P1/P7 and R16F2n/R16R2 and 1,800 bp and 1,200 bp size products were amplified in first round PCR and nested-PCR respectively. The PCR product was sequenced and compared with the reference phytoplasma sequences collected from the database (NCBI). 16S rDNA sequences of Dharwad chickpea phytoplasma shared the highest nucleotide identity of (>98%) with Periwinkle phyllody16SrII-E (EU096500). This study indicated the association of ‘Candidatus Phytoplasma aurantifolia’ the 16SrII-E group infecting chickpea from Northern Karnataka.

scholarly journals First Report on the Detection and Molecular Characterization of Bovine Herpesvirus 1 from a Clinical case of Infectious Bovine Rhinotracheitis in Pakistan

2021 ◽  
Vol 41 (01) ◽  
pp. 160-162
Author(s):  
Aayesha Riaz
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
Bovine Herpesvirus ◽  
Nasal Discharge ◽  
Infectious Bovine Rhinotracheitis ◽  
Bovine Herpesvirus 1 ◽  
Nasal Swabs ◽  
Herpesvirus 1 ◽  
First Time

Bovine herpesvirus 1 (BHV-1) is a noteworthy reason for many Cattle/Buffalo diseases. Infectious bovine rhinotracheitis (IBR) is one of the diseases which are caused by the BHV-1. In the present study a cow which was suspected of IBR was examined. The animal was suffering from fever and respiratory distress along with rhinitis (red nose), in appetence, and dyspnea. The nasal mucosa and muzzle were distinctly inflamed, with nasal discharge. DNA from blood samples and nasal swabs were subjected to nested PCR using glycoprotein B gene (gB) Primers. The samples were found positive for BHV-1 gB gene. Sequencing and phylogenetic analysis revealed close similarities with other BHV-1 gB gene sequences. The accession numbers assigned to this pioneer sequences in GenBank are MT449510 (BHV-1-Pak 1) and MT449511 (BHV-1-Pak 2). In this study, we reported for the first time the detection of DNA of BHV-1 through nested PCR assay and molecular characterization of BHV-1 gB gene in Pakistan. This study will be useful in further diagnoses of BHV-1 in Pakistan and in development of BHV-1 vaccine to reduce economical losses due to IBR

scholarly journals Apparent density, trypanosome infection rates and host preference of tsetse flies in the sleeping sickness endemic focus of   Northwestern Uganda

2020 ◽  
Author(s):  
Robert Opiro ◽  
Robert Opoke ◽  
Harriet Angwech ◽  
Esther Nakafu ◽  
Francis A. Oloya ◽  
...  
Keyword(s):  
Infection Rate ◽  
Apparent Density ◽  
Blood Meal ◽  
Nested Pcr ◽  
Tsetse Fly ◽  
Tsetse Flies ◽  
Trypanosome Species ◽  
Trypanosome Infection ◽  
Infection Rates ◽  
Monitor Lizard

Abstract Background: African trypanosomiasis, caused by protozoa of the genus Trypanosoma and transmitted by the tsetse fly, is a serious parasitic disease of humans and animals. Reliable data on the vector distribution, feeding preference and the trypanosome species they carry is pertinent to planning sustainable control strategies.Methodology: We deployed 109 biconical traps in 10 villages in two districts of northwestern Uganda to obtain information on the apparent density, trypanosome infection rates and blood meal sources of tsetse flies. A subset (272) of the collected samples was analyzed for detection of trypanosomes species and sub-species using a nested PCR protocol based on primers amplifying the Internal Transcribed Spacer (ITS) region of ribosomal DNA. 34 blood-engorged adult tsetse midguts were analyzed for blood meal sources by sequencing of the mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) genes. Results: Out of the 109 traps deployed, we captured 622 Glossina fuscipes fuscipes tsetse flies (269 males and 353 females). Apparent density (AD) ranged from 0.6 to 3.7 flies/trap/day in the two districts. 29 (10.7%) of the flies were infected with one or more trypanosome species. Infection rate was not significantly associated with neither age group (χ2 = 5.001, df=2, 0.082), sex of the fly (χ2 = 0.099, df = 1, p = 0.753), district of origin (χ2= 0.629, df = 1, p = 0.428) nor village (χ2= 9.252, df = 9, p = 0.414). Nested PCR revealed several species of trypanosomes: T. vivax (6.62%), T. congolense (2.57%), T. brucei and T. simiae each at 0.73%. Blood meal analyses revealed five principal vertebrate hosts, namely, cattle (Bos taurus), humans (Homo sapiens), Nile monitor lizard (Varanus niloticus), African mud turtle (Pelusio schapini) and the African Savanna elephant (Loxodonta africana).Conclusion: We found an infection rate of 10.78 %, with all infections attributed to trypanosome species that are causative agents for the animal disease only. However, more verification of this finding using large-scale passive and active screening of human and tsetse samples should be done. Cattle and humans appear to be the most important tsetse hosts in the region and should be considered in the design of interventions.

scholarly journals Apparent density, trypanosome infection rates and host preference of tsetse flies in the sleeping sickness endemic focus of Northwestern Uganda

2020 ◽  
Author(s):  
Robert Opiro ◽  
Robert Opoke ◽  
Harriet Angwech ◽  
Esther Nakafu ◽  
Francis A. Oloya ◽  
...  
Keyword(s):  
Infection Rate ◽  
Apparent Density ◽  
Blood Meal ◽  
Nested Pcr ◽  
Large Scale ◽  
Tsetse Fly ◽  
Tsetse Flies ◽  
Trypanosome Species ◽  
Infection Rates ◽  
Monitor Lizard

Abstract Background: African trypanosomiasis, caused by protozoa of the genus Trypanosoma and transmitted by the tsetse fly, is a serious parasitic disease of humans and animals. Reliable data on the vector distribution, feeding preference and the trypanosome species they carry is pertinent to planning sustainable control strategies.Methodology: We deployed 109 biconical traps in 10 villages in two districts of northwestern Uganda to obtain information on the apparent density, infection rates and blood meal sources of tsetse flies. A subset (272) of the collected samples was analyzed for detection of trypanosomes species and sub-species using a nested PCR protocol based on primers amplifying the Internal Transcribed Spacer (ITS) region of ribosomal DNA. 34 blood-engorged adult tsetse midguts were analyzed for blood meal sources by sequencing of the mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) genes. Results: Out of the 109 traps deployed, we captured 622 Glossina fuscipes fuscipes tsetse flies (269 males and 353 females). Apparent density (AD) ranged from 0.6 to 3.7 flies/trap/day in the two districts. 29 (10.7%) of the flies were infected with one or more trypanosome species. Infection rate was not significantly associated with neither age group (χ2 = 5.001, df=2, 0.082), sex of the fly (χ2 = 0.099, df = 1, p = 0.753), district of origin (χ2= 0.629, df = 1, p = 0.428) nor village (χ2= 9.252, df = 9, p = 0.414). Nested PCR revealed several species of trypanosomes: T. vivax (6.62%), T. congolense (2.57%), and T. brucei and T. simiae each at 0.73%. Blood meal analyses revealed five principal vertebrate hosts, namely, cattle (Bos taurus), humans (Homo sapiens), Nile monitor lizard (Varanus niloticus), African mud turtle (Pelusio schapini) and the African Savanna elephant (Loxodonta africana).Conclusion: We found a moderately high infection rate of 10.78%, with all infections attributed to trypanosome species that are causative agents for the animal disease only. However, more validation using large-scale passive and active screening of human and tsetse samples should be done. Cattle and humans appear to be the most important tsetse hosts in the region and should be considered in the design of interventions.

scholarly journals Apparent density, trypanosome infection rates and host preference of tsetse flies in the sleeping sickness endemic focus of northwestern Uganda

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Robert Opiro ◽  
Robert Opoke ◽  
Harriet Angwech ◽  
Esther Nakafu ◽  
Francis A. Oloya ◽  
...  
Keyword(s):  
Infection Rate ◽  
Apparent Density ◽  
Blood Meal ◽  
Nested Pcr ◽  
Large Scale ◽  
Tsetse Fly ◽  
Tsetse Flies ◽  
Trypanosome Species ◽  
Trypanosome Infection ◽  
Monitor Lizard

Abstract Background African trypanosomiasis, caused by protozoa of the genus Trypanosoma and transmitted by the tsetse fly, is a serious parasitic disease of humans and animals. Reliable data on the vector distribution, feeding preference and the trypanosome species they carry is pertinent to planning sustainable control strategies. Methodology We deployed 109 biconical traps in 10 villages in two districts of northwestern Uganda to obtain information on the apparent density, trypanosome infection status and blood meal sources of tsetse flies. A subset (272) of the collected samples was analyzed for detection of trypanosomes species and sub-species using a nested PCR protocol based on primers amplifying the Internal Transcribed Spacer (ITS) region of ribosomal DNA. 34 blood-engorged adult tsetse midguts were analyzed for blood meal sources by sequencing of the mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) genes. Results We captured a total of 622 Glossina fuscipes fuscipes tsetse flies (269 males and 353 females) in the two districts with apparent density (AD) ranging from 0.6 to 3.7 flies/trap/day (FTD). 10.7% (29/272) of the flies were infected with one or more trypanosome species. Infection rate was not significantly associated with district of origin (Generalized linear model (GLM), χ2 = 0.018, P = 0.895, df = 1, n = 272) and sex of the fly (χ2 = 1.723, P = 0.189, df = 1, n = 272). However, trypanosome infection was highly significantly associated with the fly’s age based on wing fray category (χ2 = 22.374, P < 0.001, df = 1, n = 272), being higher among the very old than the young tsetse. Nested PCR revealed several species of trypanosomes: T. vivax (6.62%), T. congolense (2.57%), T. brucei and T. simiae each at 0.73%. Blood meal analyses revealed five principal vertebrate hosts, namely, cattle (Bos taurus), humans (Homo sapiens), Nile monitor lizard (Varanus niloticus), African mud turtle (Pelusios chapini) and the African Savanna elephant (Loxodonta africana). Conclusion We found an infection rate of 10.8% in the tsetse sampled, with all infections attributed to trypanosome species that are causative agents for AAT. However, more verification of this finding using large-scale passive and active screening of human and tsetse samples should be done. Cattle and humans appear to be the most important tsetse hosts in the region and should be considered in the design of control interventions.

scholarly journals Apparent Density, Trypanosome Infection Rates and Host Preference of Tsetse Flies in the Sleeping Sickness Endemic Focus of North-Western Uganda

2020 ◽  
Author(s):  
Robert Opiro ◽  
Robert Opoke ◽  
Harriet Angwech ◽  
Esther Nakafu ◽  
Francis A. Oloya ◽  
...  
Keyword(s):  
Infection Rate ◽  
Apparent Density ◽  
Blood Meal ◽  
Nested Pcr ◽  
Tsetse Fly ◽  
Tsetse Flies ◽  
Trypanosome Species ◽  
Trypanosome Infection ◽  
Infection Rates ◽  
Monitor Lizard

Abstract Background: African trypanosomiasis, caused by protozoa of the genus Trypanosoma and transmitted by the tsetse fly, is a serious parasitic disease of humans and animals. Reliable data on the vector distribution, feeding preference and the trypanosome species they carry is pertinent to planning sustainable control strategies.Methodology: We deployed 109 biconical traps in 10 villages in two districts of northwestern Uganda to obtain information on the apparent density, trypanosome infection rates and blood meal sources of tsetse flies. A subset of the collected samples was analyzed for detection of trypanosomes species and sub-species using a nested PCR protocol based on primers amplifying the Internal Transcribed Spacer (ITS) region of ribosomal DNA. 34 blood-engorged adult tsetse midguts were analyzed for blood meal sources by sequencing of the mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) genes. Results: Out of the 109 traps deployed, we captured 622 Glossina fuscipes fuscipes tsetse flies (269 males and 353 females). Apparent density (AD) ranged from 0.6 to 3.7 flies/trap/day in the two districts. 29 (10.7%) of the flies were infected with one or more trypanosome species, with infection rate significantly associated with age group (χ2 = 29.733, df = 2, p < 0.05) but not with sex (χ2 = 0.43, df = 1, p = 0.835) and district of origin (χ2 = 1.374, df = 1, p = 0.241). Nested PCR revealed several species of trypanosomes: T. vivax (62.1%), T. congolense (24.14 %), and T. brucei and T. simiae each at 6.89%. Blood meal analyses revealed five principal vertebrate hosts, namely, cattle (Bos taurus), humans (Homo sapiens), Nile monitor lizard (Varanus niloticus), African mud turtle (Pelusio schapini) and the African Savanna elephant (Loxodonta africana).Conclusion: We found a moderately high infection rate at 10.78%, with all infections attributed to trypanosome species that are causative agents for the animal disease only. However, more validation using large-scale passive and active screening of human and tsetse samples should be done. Cattle and humans appear to be the most important tsetse hosts in the region and should be considered in the design of interventions.

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