scholarly journals First Molecular Evidence of Anaplasma bovis and Anaplasma phagocytophilum in Bovine from Central Punjab, Pakistan

Pathogens ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 155 ◽  
Author(s):  
Naveed Iqbal ◽  
Muhammad Uzair Mukhtar ◽  
Jifei Yang ◽  
Muhammad Sohail Sajid ◽  
Qingli Niu ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Molecular Data ◽  
Molecular Evidence ◽  
Potential Threat ◽  
Nested Polymerase Chain Reaction ◽  
Fta Cards ◽  
Causative Agents ◽  
Polymerase Chain ◽  
Obligate Intracellular Bacteria ◽  
Anaplasma Bovis

Obligate intracellular bacteria belonging to the genus Anaplasma spp. are responsible for causing a hemolytic disease called anaplasmosis in animals, as well as in humans. This study was aimed at the molecular identification and genetic analysis of responsible causative agents of anaplasmosis beyond those already reported. A survey was performed during July and August 2018 in the Jhang District, Punjab, Pakistan. Four hundred and fifty blood samples from asymptomatic, tick-infested cattle were collected on FTA cards and tested for the Anaplasma spp. presence using nested-polymerase chain reaction (PCR) methods. The 16S ribosomal RNA gene sequences generated from the positive samples were used for genetic analysis of Anaplasma spp. The nested-PCR results showed the presence of two Anaplasma spp. with an overall prevalence rate of 10.44%, where the prevalence of A. bovis and A. phagocytophilum was 7.78% and 2.66%, respectively. The study portrayed new molecular data on the prevalence of Anaplasma spp. in the studied cattle population, indicating a potential threat to the human population as well.

scholarly journals Granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Łukasz Adaszek ◽  
Anna Wilczyńska ◽  
Jerzy Ziętek ◽  
Marcin Kalinowski ◽  
Oliwier Teodorowski ◽  
...  
Keyword(s):  
Anaplasma Phagocytophilum ◽  
Polymerase Chain Reaction Test ◽  
Anaplasma Ovis ◽  
Case Presentation ◽  
Granulocytic Anaplasmosis ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Obligate Intracellular Bacteria ◽  
Chain Reaction Test ◽  
Anaplasma Bovis

Abstract Background Anaplasma are obligate intracellular bacteria and aetiological agents of tick-borne diseases of both veterinary and medical interest. The genus Anaplasma comprises six species: Anaplasma marginale, Anaplasma centrale, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma bovis and Anaplasma platys. They can infect humans, carnivores, ruminants, rodents, insectivores, birds and reptiles. The aim of this study was to present the first clinical case of granulocytic anaplasmosis in a captive ring-tailed lemur in Poland. Case presentation A 4-year-old female lemur presented anorexia, epistaxis and tick infestation. The microscopic examination of a blood smear revealed morulae in neutrophils. Polymerase chain reaction test and sequencing of obtained PCR product confirmed infection by the GU183908 Anaplasma phagocytophilum strain. Therapeutic protocol included doxycycline (2.5 mg/kg p.o., b.i.d.) for 3 weeks and the lemur recovered within 24 h. Conclusions This is the first report on granulocytic anaplasmosis in a ring-tailed lemur in Europe, indicating that A. phagocytophilum infection must also be considered in differential diagnosis in this animal species, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.

scholarly journals Application of immunofluorescence assay and nested polymerase chain reaction for query fever diagnosis in animal handlers of Puducherry, South India, and phylogenetic analysis based on IS1111 repetitive gene element

2019 ◽  
Vol 12 (11) ◽  
pp. 1769-1774 ◽  
Author(s):  
Jothimani Pradeep ◽  
Selvaraj Stephen ◽  
Balakrishnan Sangeetha ◽  
Prabakar Xavier Antony ◽  
S. Amsaveni ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Analysis ◽  
Tamil Nadu ◽  
South India ◽  
Molecular Evidence ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Gene Element ◽  
Animal Handlers

Background and Aim: Diagnosis of query fever (QF) is mostly done on the basis of serological/molecular tests, due to the stringent requirement of biosafety level-3 containment facilities for isolating Coxiella burnetii in culture. QF is an important zoonosis and is considered to be an occupational hazard to livestock handlers. This report describes our study on the serological as well as molecular evidence of QF in animal handlers from Puducherry and surrounding Tamil Nadu, from where, to the best of our knowledge, no such reports are available so far. Materials and Methods: Seventy-five animal handlers were recruited, comprising veterinarians, slaughterhouse workers, butchers, and animal attendants of various government veterinary clinics from Puducherry and surrounding areas of Tamil Nadu state. QF serology was performed to identify Phase I and Phase II immunoglobulin G antibodies to C. burnetii. Nested polymerase chain reaction (N-PCR) was carried out to detect C. burnetii DNA in buffy coat samples by targeting IS1111 gene element. N-PCR-positive samples were sequenced and phylogenetic analysis was performed using MEGA software version 10.0. Results: A total of 21 animal handlers (28.1%) were positive for either serology or PCR. PCR alone was positive in 10 (13.4%), only serology was positive in 8 (10.7%), and both serology and PCR were positive in three samples (4.0%). GenBank accession numbers were obtained for 13 N-PCR-positive samples (MG548608-MG548620). Six of our study sequences showed close similarity with the reference isolates from Bengaluru, Colombia, Brazil, France, and Iran. Conclusion: A significant percentage of QF positivity in animal handlers of this part of South India, Puducherry, warrants a prospective study with follow-up of a large number of this occupational group.

scholarly journals Molecular evidence of mixed P. vivax and P. falciparum infections in northern Islamic Republic of Iran

2004 ◽  
Vol 10 (3) ◽  
pp. 336-342 ◽  
Author(s):  
S. Zakeri ◽  
S. Mamaghani ◽  
A. A. Mehrizi ◽  
Z. Shahsavari ◽  
A. Raeisi ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Plasmodium Species ◽  
Single Species ◽  
Molecular Evidence ◽  
Islamic Republic ◽  
Mixed Species ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Islamic Republic Of Iran ◽  
Polymerase Chain

This study compared basic microscopy with molecular detection of Plasmodium species. According to thick-film microscopy, 100% of 142 malaria cases in Pars-Abad, Ardebil province, were infected with a single species, P vivax. However, nested polymerase chain reaction [PCR] detected mixed species infections of both P. vivax and P. falciparum in 7.0%. In Maz and eran province, 2/20 blood films were diagnosed with only P. falciparum and 18/20 with only P. vivax. However, nested PCR detected 17/20, 2/20 and 1/20 with P. vivax only, P. falciparum only and mixed species respectively. The unexpected presence of P. falciparum urges prompt investigation and immediate treatment of malaria cases in this region

scholarly journals Prevalence of canine coronavirus and parvovirus infections in dogs with gastroenteritis in Thailand

2012 ◽  
Vol 48 (No. 6) ◽  
pp. 163-168 ◽  
Author(s):  
K. Sakulwira ◽  
P. Vanapongtipagorn ◽  
A. Theamboonlers ◽  
K. Oraveerakul ◽  
Y. Poovorawan
Keyword(s):  
Polymerase Chain Reaction ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Encoding ◽  
Nested Polymerase Chain Reaction ◽  
Fecal Samples ◽  
Canine Coronavirus ◽  
Pcr Product ◽  
Causative Agents ◽  
Polymerase Chain

Canine coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the causative agents of gastroenteritis in dogs. Seventy fecal samples from dogs with signs of gastroenteritis (vomiting and diarrhea), twenty-five fecal samples from healthy dogs and one CPV-2 vaccine strain were amplified by semi-nested polymerase chain reaction (PCR) and semi-nested reverse transcriptase polymerase chain reaction (RT-PCR), aimed at specifically studying the gene encoding the most abundant capsid protein VP2 of CPV-2 and spike protein of CCV. The specificity of the CCV RT-PCR product was evaluated by sequencing. Positive specimens comprised 44 samples (62.8%) and 9 samples (12.8%) for CPV-2 and CCV, respectively. In nine CCV positive samples, seven displayed co-infection between CCV and CPV-2. Our CCV sequence (AF482001) showed a 94.9% nucleotide identity to CCV reported in GenBank accession number D13096. High prevalence of CCV and CPV-2 infections was found in 1–2 month- and 3–6 month-old dogs, respectively. Molecular biology of these viruses is important primarily for epidemic control and preventive measures.

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

Detection of hepatitis “C” virus in formalin-fixed liver tissue by nested polymerase chain reaction

1992 ◽  
Vol 37 (4) ◽  
pp. 310-314 ◽  
Author(s):  
Richard Sallie ◽  
Anne Rayner ◽  
Bernard Portmann ◽  
A. L. W. F. Eddleston ◽  
Roger Williams
Keyword(s):  
Polymerase Chain Reaction ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Liver Tissue ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Formalin Fixed

The fate of a genetically modifiedPseudomonasstrain and its transgene during the composting of poultry manure

2004 ◽  
Vol 50 (6) ◽  
pp. 415-421 ◽  
Author(s):  
J Guan ◽  
J L Spencer ◽  
M Sampath ◽  
J Devenish
Keyword(s):  
Polymerase Chain Reaction ◽  
Incubation Temperature ◽  
Genetically Modified ◽  
Chicken Manure ◽  
Detection Sensitivity ◽  
Plate Count ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Soil Microcosms ◽  
Polymerase Chain

The fate of the genetically modified (GM) Pseudomonas chlororaphis strain 3732 RN-L11 and its transgene (lacZ insert) during composting of chicken manure was studied using plate count and nested polymerase chain reaction (PCR) methods. The detection sensitivity of the nested PCR method was 165 copies of the modified gene per gram of moist compost or soil. Compost microcosms consisted of a 100-g mixture of chicken manure and peat, whereas soil microcosms were 100-g samples of sandy clay loam. Each microcosm was inoculated with 4 × 1010CFU of P. chlororaphis RN-L11. In controlled temperature studies, neither P. chlororaphis RN-L11 nor its transgene could be detected in compost microcosms after incubation temperature was elevated to 45 °C or above for one or more days. In contrast, in the compost microcosms incubated at 23 °C, the target organism was not detected by the plate count method after 6 days, but its transgene was detectable for at least 45 days. In compost bins, the target organism was not recovered from compost microcosms or soil microcosms at different levels in the bins for 29 days. However, the transgene was detected in 8 of the 9 soil microcosms and in only 1 of the 9 compost microcosms. The compost microcosm in which transgene was detected was at the lower level of the bin where temperatures remained below 45 °C. The findings indicated that composting of organic wastes could be used to reduce or degrade heat sensitive GM microorganisms and their transgenes.Key words: composting, genetically modified Pseudomonas strain, transgene, polymerase chain reaction.

Seasonal pattern and genotype distribution of sapovirus infection in Japan, 2003–2009

2011 ◽  
Vol 140 (1) ◽  
pp. 74-77 ◽  
Author(s):  
S. K. DEY ◽  
O. PHATHAMMAVONG ◽  
T. D. NGUYEN ◽  
A. THONGPRACHUM ◽  
W. CHAN-IT ◽  
...  
Keyword(s):  
Seasonal Pattern ◽  
Age Groups ◽  
Cold Season ◽  
Viral Gastroenteritis ◽  
Genotype Distribution ◽  
Chain Reaction ◽  
The Family ◽  
Causative Agents ◽  
Polymerase Chain ◽  
Predominant Strain

SUMMARYSapovirus, a member of the family Caliciviridae, is one of the major causative agents of viral gastroenteritis affecting all age groups. A total of 3232 faecal specimens collected from infants and children with gastroenteritis in five different regions of Japan during 2003–2009 were examined for sapovirus by reverse transcription–polymerase chain reaction. Sapoviruses were detected in 131 (4·05%) patients with the peak observed mainly in the cold season (November–March) in Japan during 2003–2009. During the last 6 years, sapovirus GI/1 was the predominant strain in Japan followed by GIV, GII/3, GII/6, GII/2, GII/12 and GI, respectively.

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