scholarly journals Molecular diagnosis of Hepatozoon canis in symptomatic dogs in the city of Goiania, Goiás, Brazil

2016 ◽  
Vol 68 (6) ◽  
pp. 1431-1439
Author(s):  
S.C. Duarte ◽  
J.A. Parente ◽  
O.J. Silveira Neto ◽  
V.S. Jayme ◽  
T.S.A. Bastos ◽  
...  
Keyword(s):  
Laboratory Diagnosis ◽  
Extraction Process ◽  
Hepatozoon Canis ◽  
Phylogenetic Study ◽  
Blood Smears ◽  
Two Samples ◽  
Pcr Products ◽  
Hepatozoon Americanum ◽  
The City ◽  
Veterinary Hospital

ABSTRACT More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.

scholarly journals Ikan teri Engraulis encrasicolus berpotensi sebagai pangan fungsional

2021 ◽  
Vol 6 (1) ◽  
pp. 75
Author(s):  
Wa Ode Salma ◽  
Ruwiah Ruwiah ◽  
Febriana Muchtar ◽  
Nabilah Hanum Mudjahidah ◽  
Karwika Dwi Saputri Nurdin ◽  
...  
Keyword(s):  
Docosahexaenoic Acid ◽  
Functional Food ◽  
Metabolic Disorders ◽  
Extraction Process ◽  
Good Prospect ◽  
Engraulis Encrasicolus ◽  
The Future ◽  
Soxhlet Method ◽  
Oil Extract ◽  
The City

Anchovy Engraulis encrasicolus is a good prospect to be developed in the future, maybe it can be used for hyperlipidemia diet.  This study, therefore, aims to measure and assess the content of EPA (eikosapentaenoat acid)  and DHA (docosahexaenoic acid).from Anchovy Engraulis encrasicolus  oil extract. The samples were obtained from the city of Kendari in Southeast Sulawesi, Indonesia. It was processed and manufactured at the Halu Oleo University in Pharmacy Laboratory. Furthermore, the extraction process utilized the Soxhlet method, while measuring and analyzing the content of EPA and DHA using UV/Vis Spectropometric. It was found that 25 mg of Anchovy Engraulis encrasicolus oil produced DHA levels of 145,5 mg/g (14,5%) and EPA levels of 97,15 mg/g (7,71%). Anchovy Engraulis encrasicolus is a potential for functional food used to provide protection against diseases associated with metabolic disorders and diet hyperlipidemia.

scholarly journals Methods for Molecular Surveillance of Influenza Used in Macedonia

PRILOZI ◽  
2014 ◽  
Vol 35 (2) ◽  
pp. 25-30
Author(s):  
Golubinka Bosevska ◽  
Elizabeta Janceska ◽  
Gordana Kuzmanovska ◽  
Vladimir Mikik ◽  
Nikola Panovski
Keyword(s):  
Influenza Virus ◽  
Predictive Value ◽  
Influenza A ◽  
Rna Extraction ◽  
Influenza Virus Infection ◽  
Laboratory Diagnosis ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Two Samples ◽  
Nose And Throat

AbstractThe aim: To present and compare different Nucleic Acid Testing assays used for laboratory diagnosis of influenza virus infection in our country.Materials and methods: Respiratory samples used were nose and throat swabs. The RNA extraction was performed with a QIAamp viral RNA kit. During the season 2009–2010 the first 25 samples were tested with: conventional gel-based RT-PCR and CDC rtRT-PCR using published specific matrix and HA gene primers and probes for influenza virus typing and subtyping.Results: Of 25 samples tested with conventional RT-PCR7(28%) were positive for influenza A, but negative for A/H1seasonal and A/H3. Retested with rtRT-PCR 9(36%) were positive for influenza A, 8(32%) were positive for A/H1pdm and 1(4%) was A/H3. Two samples positive with rtRT-PCR for influenza A were negative with RT-PCR. The sensitivity of the RT-PCR in comparison with rtRT-PCR is 100% and the specificity is 88.89%. Positive predictive value for RT-PCR is 77.78%, and negative predictive value is 100%. RT-PCR is a four-step and rtRT-PCR a one-step procedure. The turn-around time of RT-PCR is 6 hours and for rtRT-PCR it is 2 hours.Discussion and conclusion: For surveillance purposes nose and throat swabs are the more easy and practical to collect. It was proved that RT-PCR is too laborious, multi-step and time-consuming. The sensitivity of both assays is equal. The specificity of rtRT-PCR is higher. NAT assays for detection of influenza viruses have become an integral component of the surveillance programme in our country. They provide a fast, accurate and sensitive detection of influenza.

Phylogenetic analysis of DNA sequence data.

2021 ◽  
pp. 265-282
Author(s):  
Sergei A. Subbotin
Keyword(s):  
Phylogenetic Analysis ◽  
Sequence Data ◽  
Sequence Similarity ◽  
Phylogenetic Study ◽  
Public Database ◽  
Sample Collection ◽  
Minimum Evolution ◽  
Molecular Phylogenetic ◽  
Pcr Products ◽  
Selection Of

Abstract The goal of phylogenetics is to construct relationships that are true representations of the evolutionary history of a group of organisms or genes. The history inferred from phylogenetic analysis is usually depicted as branching in tree-like diagrams or networks. In nematology, phylogenetic studies have been applied to resolve a wide range of questions dealing with improving classifications and testing evolution processes, such as co-evolution, biogeography and many others. There are several main steps involved in a phylogenetic study: (i) selection of ingroup and outgroup taxa for a study; (ii) selection of one or several gene fragments for a study; (iii) sample collection, obtaining PCR products and sequencing of gene fragments; (iv) visualization, editing raw sequence data and sequence assembling; (v) search for sequence similarity in a public database; (vi) making and editing multiple alignment of sequences; (vii) selecting appropriate DNA model for a dataset; (viii) phylogenetic reconstruction using minimum evolution, maximum parsimony, maximum likelihood and Bayesian inference; (ix) visualization of tree files and preparation of tree for a publication; and (x) sequence submission to a public database. Molecular phylogenetic study requires particularly careful planning because it is usually relatively expensive in terms of the cost in reagents and time.

scholarly journals Natural hemoplasma infection of cats in Cuiaba, Mato Grosso, Brazil

2018 ◽  
Vol 39 (2) ◽  
pp. 875 ◽  
Author(s):  
Herica Makino ◽  
Daphine Ariadne Jesus de Paula ◽  
Valéria Regia Franco Sousa ◽  
Adriane Jorge Mendonça ◽  
Valéria Dutra ◽  
...  
Keyword(s):  
Blood Count ◽  
Clinical Care ◽  
Mato Grosso ◽  
Blood Samples ◽  
Factors Associated ◽  
Mixed Breed ◽  
The City ◽  
Veterinary Hospital ◽  
Candidatus Mycoplasma Turicensis

The aim of this research was to investigate natural hemoplasma infection in cats treated at the Veterinary Hospital of the Federal University of Mato Grosso, and the factors associated with infection. Blood samples from 151 cats of different sexes, breeds, and ages were analyzed by PCR and blood count. The overall occurrence of hemoplasma was 25.8%. Mycoplasma haemofelis (Mhf), ‘Candidatus Mycoplasma haemominutum (CMhm)’, and ‘Candidatus Mycoplasma turicensis’ (CMt) were observed in 15.2%, 14.6% and 2.6% of cats, respectively. In 6.6 % of cases, co-infection was observed. Male felines or mixed breed cats were associated with infection by CMhm (P = 0.02 and 0.04, respectively). The data obtained demonstrated an occurrence of 25.8% for hemoplasma infection in felines coming from clinical care in the city of Cuiabá, where males were at higher risk of acquiring the infection by these agents, in addition to a higher risk for CMhm in felines with no specific breed.

Prevalence of Paraneoplastic Syndromes in Dogs and Cats Treated in a Veterinary Hospital in the City of São Paulo, Brazil

2017 ◽  
Vol 156 (1) ◽  
pp. 114
Author(s):  
C. Andrade ◽  
R. Rodrigues ◽  
H. Stevanini ◽  
C. Leite ◽  
M. Santos ◽  
...  
Keyword(s):  
Sao Paulo ◽  
Paraneoplastic Syndromes ◽  
São Paulo ◽  
The City ◽  
Veterinary Hospital

scholarly journals Molecular detection of feline arthropod-borne pathogens in cats in Cuiabá, state of Mato Grosso, central-western region of Brazil

2013 ◽  
Vol 22 (3) ◽  
pp. 385-390 ◽  
Author(s):  
Natasha Gandolfi Miceli ◽  
Fernando Antonio Gavioli ◽  
Luiz Ricardo Gonçalves ◽  
Marcos Rogério André ◽  
Valéria Régia Franco Sousa ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Western Region ◽  
Mato Grosso ◽  
Animal Shelters ◽  
Blood Samples ◽  
Cytauxzoon Felis ◽  
The City ◽  
Veterinary Hospital

Hemotrophic mycoplasmas (hemoplasmas), Bartonellasp., Hepatozoon sp. and Cytauxzoon felis are prominent pathogens that circulate between cats and invertebrate hosts. The present study aimed to detect the presence of DNA from hemoplasmas,Bartonella sp., Hepatozoon sp. andCytauxzoon felis, and then confirm it by means of sequencing, in blood samples from cats in Cuiabá, MT, Brazil. From February 2009 to February 2011, blood samples with added EDTA were collected from 163 cats that were being housed in four different animal shelters in the city of Cuiabá, state of Mato Grosso, Brazil and from 15 cats that were admitted to the veterinary hospital of the Federal University of Mato Grosso (UFMT). Out of the 178 cats sampled, 15 (8.4%) were positive for hemoplasmas: four (2.2%) forMycoplasma haemofelis, 12 (6.7%) for ‘Candidatus M. haemominutum’ and one (0.5%) for ‘Candidatus M. turicensis’. One cat (0.5%), a patient that was attended at the veterinary hospital, was coinfected with M. haemofelis, ‘Candidatus M. haemominutum’ and ‘Candidatus M. turicensis’, based on sequencing confirmation. Four cats were positive for Bartonella spp.: three (1.7%) for B. henselae and one (0.5%) for B. clarridgeiae. None of the animals showedCytauxzoon sp. or Hepatozoon sp. DNA in their blood samples. This study showed that cats housed in animal shelters in the city of Cuiabá, state of Mato Grosso, are exposed to hemoplasmas andBartonella species.

Comparison of commercial DNA extraction kits for isolation and purification of bacterial and eukaryotic DNA from PAH-contaminated soils

2011 ◽  
Vol 57 (8) ◽  
pp. 623-628 ◽  
Author(s):  
Nagissa Mahmoudi ◽  
Greg F. Slater ◽  
Roberta R. Fulthorpe
Keyword(s):  
Dna Extraction ◽  
Contaminated Soils ◽  
Dna Isolation ◽  
Extraction Process ◽  
Organic Carbon Content ◽  
Soil Dna ◽  
Isolation And Purification ◽  
Dna Yield ◽  
Pcr Products ◽  
Eukaryotic Dna

Molecular characterization of the microbial populations of soils and sediments contaminated with polycyclic aromatic hydrocarbons (PAHs) is often a first step in assessing intrinsic biodegradation potential. However, soils are problematic for molecular analysis owing to the presence of organic matter, such as humic acids. Furthermore, the presence of contaminants, such as PAHs, can cause further challenges to DNA extraction, quantification, and amplification. The goal of our study was to compare the effectiveness of four commercial soil DNA extraction kits (UltraClean Soil DNA Isolation kit, PowerSoil DNA Isolation kit, PowerMax Soil DNA Isolation kit, and FastDNA SPIN kit) to extract pure, high-quality bacterial and eukaryotic DNA from PAH-contaminated soils. Six different contaminated soils were used to determine if there were any biases among the kits due to soil properties or level of contamination. Extracted DNA was used as a template for bacterial 16S rDNA and eukaryotic 18S rDNA amplifications, and PCR products were subsequently analyzed using denaturing gel gradient electrophoresis (DGGE). We found that the FastDNA SPIN kit provided significantly higher DNA yields for all soils; however, it also resulted in the highest levels of humic acid contamination. Soil texture and organic carbon content of the soil did not affect the DNA yield of any kit. Moreover, a liquid–liquid extraction of the DNA extracts found no residual PAHs, indicating that all kits were effective at removing contaminants in the extraction process. Although the PowerSoil DNA Isolation kit gave relatively low DNA yields, it provided the highest quality DNA based on successful amplification of both bacterial and eukaryotic DNA for all six soils. DGGE fingerprints among the kits were dramatically different for both bacterial and eukaryotic DNA. The PowerSoil DNA Isolation kit revealed multiple bands for each soil and provided the most consistent DGGE profiles among replicates for both bacterial and eukaryotic DNA.

scholarly journals Detection of heavy metals in common vegetables at Varaždin City Market, Croatia

2016 ◽  
Vol 67 (4) ◽  
pp. 340-350 ◽  
Author(s):  
Zvjezdana Stančić ◽  
Dinko Vujević ◽  
Ana Gomaz ◽  
Saša Bogdan ◽  
Dragutin Vincek
Keyword(s):  
Heavy Metals ◽  
Common Bean ◽  
Agricultural Production ◽  
Absorption Spectrometry ◽  
White Potato ◽  
Spectrometry Method ◽  
Two Samples ◽  
The Relationship ◽  
The City ◽  
Sources Of Contamination

Abstract The present study was aimed at the estimation of heavy metal content in vegetables sold at the city market of one of the densely populated Croatian cities, Varaždin, and to establish the relationship between their levels and possible sources of contamination. Twenty-eight samples of the most common diet vegetables (red and white potato, onion, carrot, common bean, lettuce, and cabbage) were randomly bought at the market in September and October 2013. Using the atomic absorption spectrometry method, concentrations of nine heavy metals (As, Cd, Cr, Cu, Hg, Mn, Ni, Pb, and Zn) were measured in the selected samples. The results showed that, in five out of 28 samples analysed, six concentrations exceeded the maximum levels provided for in the regulations: five for Pb and one for Cd. Maximum regulated levels for Pb were exceeded in two samples of red potato, two samples of common bean, and one sample of carrot (17.9 %), and for Cd in a sample of red potato (3.6 %). In conclusion, the cause of the overstepping of the maximum levels for Pb and Cd in the vegetables analysed was most likely the contaminated soil. The possible sources of soil contamination include traffic, nearby industry, floodwaters of rivers and streams, and the use of pesticides and fertilisers in agricultural production.

A Novel Heterozygous Point Mutation Causing Coagulation Factor XII Deficiency as the Molecular Mechanism of Fatal Thrombosis Event in a Chinese Pedigree.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4080-4080
Author(s):  
Ying Feng ◽  
Ye Xu ◽  
Ying Pang ◽  
Jing Dai ◽  
Wei Xie ◽  
...  
Keyword(s):  
Molecular Mechanism ◽  
Coagulation Factor ◽  
Laboratory Diagnosis ◽  
Factor Ix ◽  
The Other ◽  
Heterozygous Mutation ◽  
Factor Xii ◽  
Phenotypic Analysis ◽  
Factor Xii Deficiency ◽  
Pcr Products

Abstract Objective: To study on the laboratory diagnosis and molecular mechanism of a pedigree with fatal cerebral thrombosis event caused by hereditary coagulation factor XII deficiency. Methods: Activated partial thromboplastin time (APTT), prothrombin time (PT), coagulation factor VIII activity (FVIII:C), factor IX activity (FIX:C), factor XI and factor XII activity (FXI:C and FXII:C) were determined in the propositus and the other kindred member for phenotypic analysis. All the 14 exons and their flank sequences of factor XII gene form the propositus were amplified by PCR. The purified PCR products were sequenced directly. After mutation was found, the corresponding gene fragments covering the mutated point from the other member of the kindred were amplified and sequenced. Results: The APTT of the propositus was significantly elongated (>120 seconds), while her PT was normal. Her FVIII:C,FIX:C and FXI:C were all normal. But her FXII:C was only 1 %, lower than normal. A heterozygous G3774C missense mutation in exon 4 was identified, which has led to the substitution of serine (TCT) 73 for cysteine (TGT). The sequencing results from the pedigree suggested that the other asymptomatic member of the kindred also had the same heterozygous mutation. Conclusion: The Propositus was diagnosed with inherited coagulation factor XII deficiency. The novel gene mutation found in this pedigree is the molecular mechanism leading to the fatal thrombosis events in the propositus.

scholarly journals Hematological values associated to the serological and molecular diagnostic in cats suspected of Ehrlichia canisinfection

2013 ◽  
Vol 22 (4) ◽  
pp. 470-474 ◽  
Author(s):  
ísis Assis Braga ◽  
Luana Gabriela Ferreira dos Santos ◽  
Andréia Lima Tomé Melo ◽  
Felipe Wolf Jaune ◽  
Thaysa Felfili Ziliani ◽  
...  
Keyword(s):  
Molecular Diagnostic ◽  
Rrna Gene ◽  
Igg Antibody ◽  
Mato Grosso ◽  
Pcr Products ◽  
Antibody Titers ◽  
Polymerase Chain ◽  
Pcr Targeting ◽  
Veterinary Hospital ◽  
Hematological Alterations

The literature contains several studies on feline ehrlichiosis. However, information about the characteristics of Ehrlichiainfection in cats is still scanty. This study evaluated the association between Ehrlichia spp. infection and the hematologic data of 93 cats treated at the Federal University of Mato Grosso Veterinary Hospital in Cuiabá, state of Mato Grosso, Brazil. The presence of or exposure to Ehrlichia spp. infection was evaluated by Polymerase Chain Reaction (PCR) targeting the dsb and 16S rRNA gene of Ehrlichia, and by detection of anti-Ehrlichia canis IgG antibodies in Indirect Fluorescence Assay (IFA), respectively. Eight (8.6%) cats tested positive by PCR and the partial DNA sequence obtained from PCR products was a 100% match to E. canis. Forty-two (45.1%) cats showed antibody reactivity against Ehrlichia spp. Hematological alterations such as low erythrocyte count, thrombocytopenia, lymphopenia and monocytosis were observed in PCR positive cats. Among them, low erythrocyte counts were associated with IgG antibody titers of 40 to 640 and five cats also tested positive by PCR. Furthermore, PCR-positive cats showed a tendency to be lymphopenic. No correlation was found between age and sex, and no ticks were observed in any of the examined cats.

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