scholarly journals Natural canine infection by Leishmania infantum and Leishmania amazonensis and their implications for disease control

2016 ◽  
Vol 25 (4) ◽  
pp. 465-469 ◽  
Author(s):  
Letícia da Cruz Sanches ◽  
Cleber Costa de Martini ◽  
Alex Akira Nakamura ◽  
Maria Emília Bodini Santiago ◽  
Beatriz Dolabela de Lima ◽  
...  
Keyword(s):  
Public Health Problem ◽  
Leishmania Species ◽  
Canine Leishmaniasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rrna Gene ◽  
Major Public Health Problem ◽  
Pcr Rflp ◽  
New Hosts ◽  
Canine Infection ◽  
Polymerase Chain

Abstract Leishmaniasis is a major public health problem worldwide. Because Leishmania can adapt to new hosts or vectors, knowledge concerning the current etiological agent in dogs is important in endemic areas. This study aimed to identify the Leishmania species detected in 103 samples of peripheral blood from dogs that were naturally infected with these protozoa. The diagnosis of leishmaniasis was determined through parasitological examination, the indirect enzyme-linked immunosorbent assay (ELISA) and the polymerase chain reaction (PCR). The Leishmania species were identified by means of PCR-restriction fragment length polymorphism (PCR-RFLP). The samples were subjected to PCR using oligonucleotide primers that amplify the intergenic region ITS1 of the rRNA gene in order to identify the species. The amplified DNA was digested using the restriction enzyme HaeIII. A restriction profile identical to L. amazonensis was shown in 77/103 samples and the profile was similar to L. infantum in 17/103. However, a mixed profile was shown in 9/103 samples, which impeded species identification. In conclusion, the infection in these dogs was predominantly due to L. amazonensis, thus indicating that diagnosing of cases of canine leishmaniasis needs to be reexamined, since the causative agent identified is not restricted to L. infantum.

scholarly journals Characterization of Leishmania Species Isolated from Cutaneous Human Samples from Central Region of Syria by RFLP Analysis

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Samar Anis Al-Nahhas ◽  
Rania Magdy Kaldas
Keyword(s):  
Central Region ◽  
Public Health Problem ◽  
Rflp Analysis ◽  
Leishmania Species ◽  
Etiologic Agent ◽  
Direct Microscopic Examination ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Rflp Assay

Cutaneous leishmaniasis (CL) is an endemic disease and a public health problem in Hama governorate located in the central region of Syria. The aim of this study was to characterize Leishmania species isolated from human skin samples. A polymerase chain reaction, restriction fragment length polymorphism (PCR-RFLP) assay, was performed on skin lesion material samples from 32 patients with confirmed CL by direct microscopic examination in order to prove its usefulness and efficiency for identification of Leishmania species. Leishmania tropica (L. tropica) is confirmed as an etiologic agent of CL in this area.

scholarly journals CHARACTERIZATION OF CANINE LEISHMANIASIS BY PCR-RFLP IN CUIABA, MATO GROSSO, BRAZIL

2012 ◽  
Vol 17 (2) ◽  
Author(s):  
Arleana Do Bom Parto Ferreira Almeida ◽  
Daphine Ariadne Jesus de Paula ◽  
Valéria Dutra ◽  
Edson Moleta Colodel ◽  
Luciano Nakazato ◽  
...  
Keyword(s):  
Urban Areas ◽  
Canine Leishmaniasis ◽  
Prevention Measures ◽  
Surrounding Area ◽  
Mato Grosso ◽  
Significant Difference ◽  
Pcr Rflp ◽  
Cutaneous Form ◽  
Canine Infection

Leishmaniases are neglected zoonoses that are increasing in Brazil. The dog is considered the main reservoir of the visceral form in urban areas of Brazil and also important in maintaining the cycle of transmission of the cutaneous form in endemic areas. We used PCR-RFLP to identify the species of Leishmania involved in canine infection in Cuiaba City, Mato Grosso. Samples of bone marrow and lymph were collected from 181 dogs, of which 7.2% tested positive with indirect immunofluorescence and 24.9% using PCR-RFLP; a significant difference (p ≤ 0.05), had been possible to characterize the species Leishmania (L.) chagasi. This will aid in developing prevention measures and in the control of disease in Cuiaba and the surrounding area.

scholarly journals Etiology of Diarrhea Requiring Hospitalization in Bangladesh by Quantitative Polymerase Chain Reaction, 2014–2018

2020 ◽  
Author(s):  
Mami Taniuchi ◽  
Kamrul Islam ◽  
Md Abu Sayeed ◽  
James A Platts-Mills ◽  
Md Taufiqul Islam ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Coefficient Of Variation ◽  
Quantitative Polymerase Chain Reaction ◽  
Age Groups ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Surveillance Systems ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background Diarrhea remains a major public health problem and characterization of its etiology is needed to prioritize interventions. However, most data are from single-site studies of children. We tested samples from participants of any age from 11 geographically diverse hospitals in Bangladesh to describe pathogen-specific burdens of diarrhea. Methods We utilized 2 existing diarrhea surveillance systems: a Nationwide network at 10 sentinel hospitals and at the icddr,b hospital. We tested stools from enrolled participants and nondiarrheal controls for enteropathogens using quantitative polymerase chain reaction and calculated pathogen-specific attributable fractions (AFs) of diarrhea. Results We analyzed 5516 patients with diarrhea and 735 controls. Overall, rotavirus had the highest attributable burden of diarrhea (Nationwide AF, 17.7%; 95% confidence interval [CI], 14.3–20.9%; icddr,b AF, 39.9%; 38.0–41.8%), followed by adenovirus 40/41 (Nationwide AF, 17.9%; 95% CI: 13.9–21.9%; icddr,b AF, 16.6%; 95% CI, 14.4–19.4%) and Vibrio cholerae (Nationwide AF, 10.2%; 95% CI, 9.1–11.3%; icddr,b AF, 13.3%; 95% CI: 11.9–15.1%). Rotavirus was the leading pathogen in children <5 years and was consistent across the sites (coefficient of variation = 56.3%). Adenovirus 40/41 was the second leading pathogen in both children and adults. Vibrio cholerae was the leading pathogen in individuals >5 years old, but was more geographically variable (coefficient of variation = 71.5%). Other attributable pathogens included astrovirus, norovirus, Shigella, Salmonella, ETEC, sapovirus, and typical EPEC. Conclusions Rotavirus, adenovirus 40/41, and V. cholerae were the leading etiologies of infectious diarrhea requiring hospitalization in Bangladesh. Other pathogens were important in certain age groups or sites.

scholarly journals ANIMAL MODELS FOR THE STUDY OF LEISHMANIASIS IMMUNOLOGY

2014 ◽  
Vol 56 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Elsy Nalleli Loria-Cervera ◽  
Fernando Jose Andrade-Narvaez
Keyword(s):  
Animal Models ◽  
Human Disease ◽  
Public Health Problem ◽  
Leishmania Species ◽  
Experimental Models ◽  
Sand Fly ◽  
Leishmania Infection ◽  
Major Public Health Problem ◽  
Wild Rodents ◽  
Experimental Conditions

Leishmaniasis remains a major public health problem worldwide and is classified as Category I by the TDR/WHO, mainly due to the absence of control. Many experimental models like rodents, dogs and monkeys have been developed, each with specific features, in order to characterize the immune response to Leishmania species, but none reproduces the pathology observed in human disease. Conflicting data may arise in part because different parasite strains or species are being examined, different tissue targets (mice footpad, ear, or base of tail) are being infected, and different numbers (“low” 1×102 and “high” 1×106) of metacyclic promastigotes have been inoculated. Recently, new approaches have been proposed to provide more meaningful data regarding the host response and pathogenesis that parallels human disease. The use of sand fly saliva and low numbers of parasites in experimental infections has led to mimic natural transmission and find new molecules and immune mechanisms which should be considered when designing vaccines and control strategies. Moreover, the use of wild rodents as experimental models has been proposed as a good alternative for studying the host-pathogen relationships and for testing candidate vaccines. To date, using natural reservoirs to study Leishmania infection has been challenging because immunologic reagents for use in wild rodents are lacking. This review discusses the principal immunological findings against Leishmania infection in different animal models highlighting the importance of using experimental conditions similar to natural transmission and reservoir species as experimental models to study the immunopathology of the disease.

scholarly journals Hepatozoon canis and Leishmania spp. coinfection in dogs diagnosed with visceral leishmaniasis

2016 ◽  
Vol 25 (4) ◽  
pp. 450-458 ◽  
Author(s):  
Fernanda Nazaré Morgado ◽  
Amanda dos Santos Cavalcanti ◽  
Luisa Helena de Miranda ◽  
Lúcia Helena O’Dwyer ◽  
Maria Regina Lucas da Silva ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Dna Isolation ◽  
Parasite Burden ◽  
Leishmania Species ◽  
18S Rrna Gene ◽  
Hepatozoon Canis ◽  
Histopathological Analysis ◽  
Rrna Gene ◽  
Pcr Rflp ◽  
Leishmania Spp

Abstract This study describes the occurrence of dogs naturally co-infected with Hepatozoon canis and two Leishmania species: L. infantum or L. braziliensis. Four dogs serologically diagnosed with Visceral Leishmaniasis were euthanized. Liver and spleen samples were collected for histopathological analysis and DNA isolation. H. canis meronts were observed in tissues from all four dogs. H. canis infection was confirmed by PCR followed by sequencing of a fragment of 18S rRNA gene. Leishmania detection and typing was confirmed by ITS1' PCR-RFLP and parasite burden was calculated using ssrRNA quantitative qPCR. A DPP - Dual Path platform test was performed. One out (Dog #2) of four animals was asymptomatic. Dogs #1 and #4 were infected by L. infantum and were DPP test positive. Dogs #2 and #3 were infected by L. braziliensis and were DPP test negative. Furthermore, visceral dissemination was observed in Dogs #2 and #3, since L. braziliensis was detected in liver and spleen samples. The visceral dissemination of L. braziliensis associated with systemic signs suggested that this co-infection could influence the parasite burden and disease progression.

Prevalence of porA pseudogene deletion among Neisseria gonorrhoeae isolates referred to the UK’s Gonococcal Resistance to Antimicrobials Surveillance Program

Sexual Health ◽  
2017 ◽  
Vol 14 (4) ◽  
pp. 392 ◽  
Author(s):  
Martina Toby ◽  
Pamela Saunders ◽  
Michelle Cole ◽  
Vlad Grigorjev ◽  
Sarah Alexander ◽  
...  
Keyword(s):  
Neisseria Gonorrhoeae ◽  
False Negative ◽  
Public Health Problem ◽  
Surveillance Program ◽  
Major Public Health Problem ◽  
Negative Results ◽  
False Negative Results ◽  
Confirmatory Tests ◽  
Polymerase Chain ◽  
Low Prevalence

porA pseudogene-negative Neisseria gonorrhoeae isolates produce false-negative results when examined by polymerase chain reaction (PCR) with porA pseudogene targets. In the present study, 533 representative gonococcal isolates received in 2011 via the Gonococcal Resistance to Antimicrobials Surveillance Program were examined to determine the prevalence of porA-negative isolates. Less than 0.4% (2/533) of isolates were found to be reproducibly negative with the porA real-time PCR but were confirmed as N. gonorrhoeae with molecular, biochemical and immunological confirmatory tests. Sequencing revealed both isolates contained the Neisseria meningitidis porA gene. Low prevalence indicates that although these isolates do not present a major public health problem, microbiologists should remain vigilant.

scholarly journals COMPARISON OF A DNA BASED PCR APPROACH WITH CONVENTIONAL METHODS FOR THE DETECTION OF MYCOBACTERIUM TUBERCULOSIS IN MOROCCO

2012 ◽  
Vol 4 (1) ◽  
pp. e2012049 ◽  
Author(s):  
Fathiah Zakham ◽  
Oufae Lahlou ◽  
Mohammed Akrim ◽  
Nada Bouklata ◽  
Sanae Jaouhari ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Insertion Sequence ◽  
Clinical Manifestations ◽  
Public Health Problem ◽  
High Specificity ◽  
Major Public Health Problem ◽  
Predictive Values ◽  
Polymerase Chain ◽  
Pcr Techniques ◽  
Potential Tool

Background: Worldwide, tuberculosis (TB) is a major public health problem and the rapid diagnosis and appropriate chemotherapy become the first priority and a serious challenge to improve TB treatment.In the objective of early TB diagnosis and rapid detection of Mycobacterium tuberculosis (MTB) in the clinical specimens, the utility of the Polymerase Chain Reaction (PCR) using the Insertion Sequence 6110 (IS6110) as target was compared to conventional methods.Methods: Out of 305 patients with different clinical manifestations: suspected, new, drug relapse, drug failure and chronic cases were enrolled in this study and tested by mycobacteriological and PCR techniques for the investigation about the tubercle bacilli.Results: The results of the in house IS6110 PCR showed a good sensitivity (92, 42%) and high specificity (98%), the positive and negative predictive values were 96.4 % and 95.3 % respectively.Conclusion: This study showed clearly that the PCR testing using the IS6110 in the routine analysis is a potential tool for the rapid TB diagnosis, especially for critical cases and would be of great interest to help the clinician in the misdiagnosed critical cases by the traditional radiology.

scholarly journals Plasmodium ovale: Parasite and Disease

2005 ◽  
Vol 18 (3) ◽  
pp. 570-581 ◽  
Author(s):  
William E. Collins ◽  
Geoffrey M. Jeffery
Keyword(s):  
Public Health Problem ◽  
Molecular Techniques ◽  
Sub Saharan Africa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Developmental Cycle ◽  
Major Public Health Problem ◽  
Plasmodium Ovale ◽  
Malaria Parasites ◽  
Sub Saharan

SUMMARY Humans are infected by four recognized species of malaria parasites. The last of these to be recognized and described is Plasmodium ovale. Like the other malaria parasites of primates, this parasite is only transmitted via the bites of infected Anopheles mosquitoes. The prepatent period in the human ranges from 12 to 20 days. Some forms in the liver have delayed development, and relapse may occur after periods of up to 4 years after infection. The developmental cycle in the blood lasts approximately 49 h. An examination of records from induced infections indicated that there were an average of 10.3 fever episodes of ≥101°F and 4.5 fever episodes of ≥104°F. Mean maximum parasite levels were 6,944/μl for sporozoite-induced infections and 7,310/μl for trophozoite-induced infections. Exoerythrocytic stages have been demonstrated in the liver of humans, chimpanzees, and Saimiri monkeys following injection of sporozoites. Many different Anopheles species have been shown to be susceptible to infection with P. ovale, including A. gambiae, A. atroparvus, A. dirus, A. freeborni, A. albimanus, A. quadrimaculatus, A. stephensi, A. maculatus, A. subpictus, and A. farauti. An enzyme-linked immunosorbent assay has been developed to detect mosquitoes infected with P. ovale using a monoclonal antibody directed against the circumsporozoite protein. Plasmodium ovale is primarily distributed throughout sub-Saharan Africa. It has also been reported from numerous islands in the western Pacific. In more recent years, there have been reports of its distribution on the Asian mainland. Whether or not it will become a major public health problem there remains to be seen. The diagnosis of P. ovale is based primarily on the characteristics of the blood stages and its differentiation from P. vivax. The sometimes elliptical shape of the infected erythrocyte is often diagnostic when combined with other, subtler differences in morphology. The advent of molecular techniques, primarily PCR, has made diagnostic confirmation possible. The development of techniques for the long-term frozen preservation of malaria parasites has allowed the development diagnostic reference standards for P. ovale. Infections in chimpanzees are used to provide reference and diagnostic material for serologic and molecular studies because this parasite has not been shown to develop in other nonhuman primates, nor has it adapted to in vitro culture. There is no evidence to suggest that P. ovale is closely related phylogenetically to any other of the primate malaria parasites that have been examined.

Molecular Detection of Hepatitis C Virus amongst Patients in Five Selected Hospitals in Niger State, Nigeria

2019 ◽  
Vol 2 (1) ◽  
pp. 60-69
Author(s):  
M U Iduh ◽  
F A Kuta ◽  
M E Abalaka ◽  
K O Shitu
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Direct Sequencing ◽  
Rna Extraction ◽  
Public Health Problem ◽  
Developed Countries ◽  
Response To Treatment ◽  
Hcv Rna ◽  
Major Public Health Problem ◽  
Polymerase Chain

Hepatitis C Virus (HCV) is a major public health problem in developing and developed countries worldwide. It is responsible for liver diseases and hepatocellular carcinoma in chronically-infected patients. This study therefore aimed to identify the strain of HCV among HCV seropositive subjects in Niger State. A total of 44 HCV seropositive blood samples which consisted of 27 males and 17 females were analyzed (after Viral RNA extraction) for the presence of HCV-RNA by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Nine (20.5%) of the samples were positive for HCV RNA. HCV-RNA positive samples were genotyped by direct sequencing at 5’UTR region genomes; sequences were aligned on MEGA 6.0 and confirmed by phylogenetic analysis. HCV genotype 1b was the only one distributed among the participants. The findings are relevant as predictors for using antiviral therapy in this population because the response to treatment varies according to the genotype.

An intracellular symbiont and other microbiota associated with field-collected populations of sawflies (Hymenoptera: Symphyta)

2008 ◽  
Vol 54 (9) ◽  
pp. 758-768 ◽  
Author(s):  
Robert I. Graham ◽  
Viviane Zahner ◽  
Christopher J. Lucarotti
Keyword(s):  
Fragment Length Polymorphism ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Intracellular Symbiont ◽  
Mountain Ash ◽  
Pcr Rflp ◽  
Gradient Gel Electrophoresis ◽  
Total Dna ◽  
Polymerase Chain

Six species of sawfly (Hymenoptera: Symphyta) from four taxonomic families (Agridae, Diprionidae, Pamphiliidae, and Tenthredinidae) were collected from locations across Canada and surveyed for their associated microbiota. Total DNA was extracted from individual insects, and polymerase chain reaction (PCR) was used to amplify the conserved 16S rRNA gene from microbiota. Denaturing gradient gel electrophoresis (DGGE) and restriction fragment length polymorphism (RFLP) were undertaken to separate bacterial clones associated with the host insect. Sequencing of the PCR–DGGE and PCR–RFLP products revealed a dominance of α- and γ-Proteobacteria, with most sequences showing high similarity to bacteria previously identified from other insect species and environmental samples. Additionally, a strain of the bacterial endosymbiont Wolbachia and a Wolbachia bacteriophage were identified from the mountain ash sawfly ( Pristiphora geniculata ).

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