scholarly journals Development of a qPCR to diagnose the genus Eimeria in bovines

2020 ◽  
Vol 41 (6) ◽  
pp. 2695-2702
Author(s):  
Sérgio Tosi Cardim ◽  
◽  
Stefani Lino Cardim ◽  
Mércia de Seixas ◽  
Alessandra Taroda ◽  
...  
Keyword(s):  
18S Rrna ◽  
High Sensitivity ◽  
Sybr Green ◽  
Morphological Observation ◽  
Economic Losses ◽  
Routine Diagnosis ◽  
Eimeria Spp ◽  
Diagnostic Training ◽  
Genus Eimeria

Bovine coccidiosis is caused by protozoa of the genus Eimeria. These protozoa mainly affect young animals, causing a decrease in production and consequent economic losses. The routine diagnosis is made through morphological observation of the oocysts, which has several limitations. The objective of the present study was to develop a qPCR technique for the diagnose of Eimeria spp. in cattle. For this purpose, the 18S rRNA region of the DNA of these parasites was selected, since it is a region with low variability among the species. The qPCR was developed using the SYBR Green, resulting in a PCR with a high sensitivity, able to amplify samples containing only one oocyst of Eimeria spp. of bovines. The feasibility of using qPCR in the diagnosis of the Eimeria Genus is demonstrated in this study, once this technique shows to be less laborious and needs less skills for diagnostic training when compared to the technique conventionally used in theroutine (micromorphometry).

scholarly journals The Mechanism Underlying the Extreme Sensitivity of Duck to Aflatoxin B1

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Kuntan Wu ◽  
Minjie Liu ◽  
Huanbin Wang ◽  
Shahid Ali Rajput ◽  
Yajing Shan ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Aflatoxin B1 ◽  
High Sensitivity ◽  
Metabolic Activation ◽  
Cytochrome P450s ◽  
Economic Losses ◽  
Metabolic Detoxification ◽  
Glutathione S Transferases ◽  
Extreme Sensitivity ◽  
Mutual Conversion

Most metabolites of aflatoxin B1 (AFB1), especially exo-AFB1-8,9-epoxide (AFBO), can induce the production of reactive oxygen species (ROS) to vary degrees, causing oxidative stress and liver damage, and ultimately induce liver cancer in humans and animals. Duck is one of the most sensitive animals to AFB1, and severe economic losses are caused by duck AFB1 poisoning every year, but the exact mechanism of this high sensitivity is still unclear. This review highlights significant advances in our understanding of the AFB1 metabolic activation, like cytochrome P450s (CYPs), and AFB1 metabolic detoxification, like glutathione S-transferases (GSTs) in poultry. In addition, AFB1 may have other metabolic pathways in poultry, such as the mutual conversion of AFB1 and aflatoxicol (AFL) and the process of AFBO to produce AFB1-8,9-dihydrodiol (AFB1-dhd) and further metabolize it into detoxification substances. This review also summarized some exogenous regulatory substances that can alleviate AFB1-induced oxidative stress.

scholarly journals Development of a Directly Visualized Recombinase Polymerase Amplification–SYBR Green I Method for the Rapid Detection of African Swine Fever Virus

2020 ◽  
Vol 11 ◽  
Author(s):  
Shuai Zhang ◽  
Aijun Sun ◽  
Bo Wan ◽  
Yongkun Du ◽  
Yanan Wu ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Sybr Green ◽  
Fever Virus ◽  
Diagnostic Methods ◽  
Sybr Green I ◽  
Recombinase Polymerase Amplification ◽  
Economic Losses ◽  
Field Samples ◽  
Standard Plasmid

African swine fever (ASF) is a lethal disease in swine caused by etiologic African swine fever virus (ASFV). The global spread of ASFV has resulted in huge economic losses globally. In the absence of effective vaccines or drugs, pathogen surveillance has been the most important first-line intervention to prevent ASF outbreaks. Among numerous diagnostic methods, recombinase polymerase amplification (RPA)-based detection is capable of producing sensitive and specific results without relying on the use of expensive instruments. However, currently used gene-specific, probe-based RPA for ASFV detection is expensive and time-consuming. To improve the efficiency of ASFV surveillance, a novel directly visualized SYBR Green I-staining RPA (RPAS) method was developed to detect the ASFV genome. SYBR Green I was added to the amplified RPA products for direct visualization by the naked eye. The sensitivity and specificity of this method were confirmed using standard plasmid and inactivated field samples. This method was shown to be highly specific with a detection limit of 103 copies/μl of ASFV in 15 min at 35°C without any cross-reactions with other important porcine viruses selected. In summary, this method enables direct sample visualization with reproducible results for ASFV detection and hence has the potential to be used as a robust tool for ASF prevention and control.

scholarly journals Sensitive SYBR Green—Real Time PCR for the Detection and Quantitation of Avian Rotavirus A

2018 ◽  
Vol 6 (1) ◽  
pp. 2 ◽  
Author(s):  
David De la Torre ◽  
Claudete Astolfi-Ferreira ◽  
Ruy Chacon ◽  
Antonio Piantino Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sybr Green ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rotavirus A ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R2 value of 0.999 and a slope of −3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%.

scholarly journals Recent Advances in Electrochemiluminescence Sensors for Pathogenic Bacteria Detection

Micromachines ◽  
2019 ◽  
Vol 10 (8) ◽  
pp. 532 ◽  
Author(s):  
Jinjin Shen ◽  
Ting Zhou ◽  
Ru Huang
Keyword(s):  
Pathogenic Bacteria ◽  
Early Stage ◽  
Point Of Care ◽  
Health And Safety ◽  
High Sensitivity ◽  
Economic Losses ◽  
Bacteria Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recent Advances ◽  
Pathogenic Bacteria Detection

Pathogenic bacterial contamination greatly threats human health and safety. Rapidly biosensing pathogens in the early stage of infection would be helpful to choose the correct drug treatment, prevent transmission of pathogens, as well as decrease mortality and economic losses. Traditional techniques, such as polymerase chain reaction and enzyme-linked immunosorbent assay, are accurate and effective, but are greatly limited because they are complex and time-consuming. Electrochemiluminescence (ECL) biosensors combine the advantages of both electrochemical and photoluminescence analysis and are suitable for high sensitivity and simple pathogenic bacteria detection. In this review, we summarize recent advances in ECL sensors for pathogenic bacteria detection and highlight the development of paper-based ECL platforms in point of care diagnosis of pathogens.

Haematobium irritans and Haematobium titillans as potential vectors of Parabronema skrjabini in camels (Camelus bactrianus) in Inner Mongolia, China

Parasitology ◽  
2020 ◽  
Vol 147 (13) ◽  
pp. 1509-1514
Author(s):  
Yang Liu ◽  
Zhiguo Zhao ◽  
Xiaoye Yang ◽  
Lianru Yang ◽  
Bo Yang ◽  
...  
Keyword(s):  
Inner Mongolia ◽  
Potential Role ◽  
Animal Husbandry ◽  
Morphological Observation ◽  
Economic Losses ◽  
Future Research ◽  
Cytochrome Oxidase Subunit ◽  
Nematode Larvae ◽  
History Of ◽  
Horn Flies

AbstractParabronema skrjabini is one of the most harmful nematodes to camels and is responsible for economic losses in animal husbandry industry. There is an urgent need for in-depth studies of potential vectors of the nematode due to its scant regarding information. As previous studies indicated that flies may be the vectors of P. skrjabini, we captured flies in the main camel-producing areas of Inner Mongolia. After autopsy of the specimens of two species of horn flies, we observed the morphology of the suspected nematode larvae found in them. Internal transcribed spacer ribosomal-DNA gene sequences were considered the best candidate to confirm the species of the larvae found. Our results showed that the homology compared with P. skrjabini was 99.5% in GenBank. Subsequently, we preliminarily identified two species of horn flies through morphological observation and then sequenced the mitochondrial-DNA-gene cytochrome oxidase subunit I obtained from two species of horn flies, with 100 and 99.2% similarity to sequences deposited in GenBank, respectively. Thus, we identified Haematobia titillans and Haematobia irritans and provided evidence for their potential role as vectors of parabronemosis. Our study provides reference for future research on the life history of the nematode and the vectors of parabronemosis.

scholarly journals Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

2015 ◽  
Vol 81 (9) ◽  
pp. 3039-3048 ◽  
Author(s):  
Stefanie Rettcher ◽  
Felicitas Jungk ◽  
Christoph Kühn ◽  
Hans-Joachim Krause ◽  
Greta Nölke ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Plant Pathogens ◽  
High Sensitivity ◽  
Potato Virus X ◽  
Plant Viruses ◽  
Economic Losses ◽  
Virus Concentration ◽  
Content Type ◽  
Agricultural Industry ◽  
Sum Frequency

ABSTRACTPlant pathogens cause major economic losses in the agricultural industry because late detection delays the implementation of measures that can prevent their dissemination. Sensitive and robust procedures for the rapid detection of plant pathogens are therefore required to reduce yield losses and the use of expensive, environmentally damaging chemicals. Here we describe a simple and portable system for the rapid detection of viral pathogens in infected plants based on immunofiltration, subsequent magnetic detection, and the quantification of magnetically labeled virus particles.Grapevine fanleaf virus(GFLV) was chosen as a model pathogen. Monoclonal antibodies recognizing the GFLV capsid protein were immobilized onto immunofiltration columns, and the same antibodies were linked to magnetic nanoparticles. GFLV was quantified by immunofiltration with magnetic labeling in a double-antibody sandwich configuration. A magnetic frequency mixing technique, in which a two-frequency magnetic excitation field was used to induce a sum frequency signal in the resonant detection coil, corresponding to the virus concentration within the immunofiltration column, was used for high-sensitivity quantification. We were able to measure GFLV concentrations in the range of 6 ng/ml to 20 μg/ml in less than 30 min. The magnetic immunoassay could also be adapted to detect other plant viruses, includingPotato virus XandTobacco mosaic virus, with detection limits of 2 to 60 ng/ml.

scholarly journals Occurrence and diversity of Eimeria species in cattle in Hamedan province, Iran

2014 ◽  
Vol 59 (No. 6) ◽  
pp. 271-275 ◽  
Author(s):  
H. Heidari ◽  
Z. Sadeghi-Dehkordi ◽  
R. Moayedi ◽  
J. Gharekhani
Keyword(s):  
Eimeria Species ◽  
Clinical Signs ◽  
Cross Sectional Study ◽  
Economic Losses ◽  
Cross Sectional ◽  
Western Iran ◽  
Eimeria Spp ◽  
Hamedan Province ◽  
Cattle Farms ◽  
Eimeria Infection

Information on the occurrence, diversity and economic losses due to Eimeria infection on cattle farms is lacking in Western Iran. The principal aim of this study was to determine the prevalence and diversity of Eimeria species in cattle in Hamedan province, Western Iran. In a cross-sectional study performed from June 2010 to April 2013, 400 faecal samples were collected randomly from animals without clinical signs from different cattle farms of Hamedan province and examined for Eimeria species infection using parasitology methods. The overall infection rate of Eimeria species was 8.25%. Nine species of Eimeria were identified including: E. bovis (23.7%), E. zuernii (19.2%), E. canadensis (12.6%), E. ellipsodalis (11.4%), E. alabamensis (10.4%), E. pellita (9.1%), E. auburnensis (6.8%), E. cylindrica (4.6%) and E. bukidnonensis (2.3%). There were statistically significant differences with respect to Eimeria infection and age group (P < 0.0001, OR = 6), although no statistically significant relationships were found for sex (P = 0.35) or faecal consistency (P = 0.587). In conclusion, this study is the first to report the prevalence and diversity of Eimeria species in cattle in Hamedan province. Infection with pathogenic Eimeria spp. was asymptomatic in all animals; this is the reason for the transmission of infection by carriers in this region. Therefore, integrated strategies should be utilised to prevent and control Eimeria spp. infection on cattle farms.  

Etio-Epidemiological Studies on Diarrhoea in Cattle and Buffalo Calves

2020 ◽  
Author(s):  
S K Sharma ◽  
Monika Joshi
Keyword(s):  
Winter Season ◽  
Epidemiological Studies ◽  
Economic Losses ◽  
Buffalo Calves ◽  
Salmonella Spp ◽  
E Coli ◽  
Cryptosporidium Spp ◽  
Faecal Samples ◽  
Eimeria Spp ◽  
Causative Agents

Calf diarrhoea is the most commonly encountered disease syndrome and significant cause of economic losses in dairy industry. Present investigation was undertaken to find out the prevalence of causative agents of diarrhoea in the bovine calves for a period of one year. The effect of age, sex, season and parity of dam was also studied. E. coli was the major organism (86.00 %) observed in the faecal samples of the diarrhoeic calves followed by rotavirus, Eimeria spp. and Amphistomes (15.00 % each); Toxocara spp. (12.00 %); Strongyles (9.00 %); Cryptosporidium spp. (6.00 %); Trichuris spp. (5.00 %); and Salmonella spp. and Strongyloides spp. (3.00 % each). The prevalence of rotavirus, Cryptosporidium spp. and Eimeria spp. was found significantly higher in buffalo calves and crossbred calves than cow calves and Gir / local non-descript calves, respectively. The prevalence of Toxocara spp., Amphistomes and Strongyles in diarrhoeic buffalo calves was significantly higher than cow calves. Highest prevalence of E. coli and rotavirus was observed in faecal samples of diarrhoeic calves of 0-15 days age group. Rotavirus was not detected in faecal samples of diarrhoeic calves above 60 days age. The susceptibility of bovine calves for E. coli and rotavirus was found decreased with the advancement of the age. The prevalence of Salmonella spp. in diarrhoeic faecal samples of bovine calves was observed only in 16-60 days age whereas Cryptosporidium spp. was found only in 0-30 days age. The most of the parasitic infestations were observed after 30 days of age in calves. The calves of both sexes were equally susceptible to different causative agents of diarrhoea. The prevalence of E. coli and most of the helminth ova in the faecal samples of diarrhoeic calves was found maximum during rainy season whereas the rotavirus was observed mostly during winter season. The prevalence of E. coli, Salmonella spp., rotavirus and Cryptosporidium spp. was found highest in the faecal samples of the diarrhoeic calves of first or second parity dams.

Improved Detection of Hg2+ in Aqueous Solution with High Sensitivity and Selectivity by Using Mercury-Specific DNA, Sybr Green I and Gold Nanoparticles

2011 ◽  
Vol 239-242 ◽  
pp. 934-939
Author(s):  
Hui Xu ◽  
Shuli Gao ◽  
Jian Nong Chen ◽  
Quan Wen Liu
Keyword(s):  
Gold Nanoparticles ◽  
High Sensitivity ◽  
Sybr Green ◽  
Sybr Green I ◽  
Fluorescence Signal ◽  
Potassium Ions ◽  
Label Free ◽  
Turn On ◽  
Sensitivity And Selectivity ◽  
Fluorescence Turn On

We report a label-free, fast, fluorescence turn on assay for Hg2+detecton by using mercury-specific DNA (MSD), Sybr Green I (SG) and gold nanoparticles (AuNPs). SG efficiently discriminates MSD and MSD/Hg2+complex. The addition of gold nanoparticle decreases the background fluorescence signal further for MSD. The fluorescence intensity of MSD/Hg2+complex keeps constant after addition of AuNPs. This property improves the signal-to-background ratio and decreases the detection limitation further. In addition, the method shows improved selectivity compared with that in the absence of AuNPs. This strategy could be applied to the detection of potassium ions and showed good generality.

scholarly journals Occurrence of poultry coccidiosis in different management systems in Thrissur, Kerala

2021 ◽  
Vol 52 (3) ◽  
Author(s):  
Karthika R. ◽  
Karthika R. ◽  
Karthika R. ◽  
Karthika R. ◽  
Karthika R. ◽  
...  
Keyword(s):  
Eimeria Species ◽  
Management Systems ◽  
Occurrence Rate ◽  
Economic Losses ◽  
Morphological Examination ◽  
Backyard Poultry ◽  
Faecal Samples ◽  
Eimeria Spp ◽  
Morphology And Morphometry ◽  
Poultry Coccidiosis

Coccidiosis is one of the most prevalent and economically important parasitic diseases caused by the infection with Eimeria species contributing to major economic losses of poultry industry worldwide. In this study, occurrence of Eimeria spp. in chicken reared under different management systems was studied. A total of 300 faecal samples from chicken were collected from six organised poultry farms and six backyard poultry units in and around Thrissur, Kerala. Out of this, 167 faecal samples were from organised farms and 133 from backyard poultry units. All the samples were artificially sporulated and examined for studying the oocysts morphology and morphometry. Out of 167 samples from organised farms 52 were found to be positive for Eimeria spp. while 61 out of 133 samples from backyard poultry were positive. The overall occurrence of Eimeria spp. in chicken from 12 different areas in and around Thrissur was 37.66 per cent (113/300). The species of Eimeria identified on morphological examination were E. tenella, E. necatrix and E. maxima. The occurrence rate of E. tenella was found to be significantly higher (46.01 %) compared to E. necatrix (39.82 %) and E. maxima (14.15 %). The rate of occurrence of Eimeria spp. infection was significantly higher in backyard poultry (45.86 %) compared to that in organised farms (31.13 %).

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