scholarly journals Bacteriological investigation of pyometra of Black Bengal goats obtained at slaughter

2019 ◽  
Vol 36 (1-2) ◽  
pp. 1-7
Author(s):  
ABMJ Uddin ◽  
M Nooruzzaman ◽  
T Akter ◽  
MAHNA Khan ◽  
MM Hossain
Keyword(s):  
Bacterial Species ◽  
Capra Hircus ◽  
Mixed Infections ◽  
Bacillus Sp ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Bacteriological Investigation ◽  
Identification Of Bacteria ◽  
Polymerase Chain ◽  
Phosphate Buffered Saline

Nine uteri of Black Bengal goats (Capra hircus) affected with pyometra were collected from three slaughterhouses at Kishoreganj district of Bangladesh. Both horns of each uterus were washed with phosphate buffered saline for isolation and identification of bacteria and its load. The bacterial loads in the uterus were high, ranging between 1 × 107 and 2.8 × 107. Six different bacterial species were identified and confirmed by Polymerase Chain Reaction (PCR). There were five Escherichia coli, six Streptococcus sp., five Staphylococcus sp, one Salmonella sp., one Pasteurella sp. and one Bacillus sp. All had mixed infections containing two or three types of bacterial pathogens. Further studies are needed for the virulence determination and antibiogram profiles. The Bangladesh Veterinarian (2019) 36(1 - 2): 1-7

scholarly journals Development of a Genoserotyping Method for Salmonella Infantis Detection on the Basis of Pangenome Analysis

2020 ◽  
Vol 9 (1) ◽  
pp. 67
Author(s):  
Seung-Min Yang ◽  
Jiwon Baek ◽  
Eiseul Kim ◽  
Hyeon-Be Kim ◽  
Seyoung Ko ◽  
...  
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Cost Effective ◽  
The Other ◽  
Gene Marker ◽  
Diagnostic Assay ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Substantial Economic Burden

In recent years, Salmonella Infantis has become a predominant serovariant in clinical and poultry isolates, thereby imposing a substantial economic burden on both public health and the livestock industry. With the aim of coping with the steep increase in serovar Infantis prevalence, a polymerase chain reaction (PCR)-based rapid and accurate diagnostic assay was developed in this study through pangenome profiling of 60 Salmonella serovars. A gene marker, SIN_02055, was identified, which is present in the S. Infantis genome but not in the pangenome of the other serovars. Primers specific to SIN_02055 were used to accurately detect serovar Infantis, and to successfully differentiate Infantis from the other 59 serovars in real-time PCR with a R2 of 0.999 and an efficiency of 95.76%. The developed method was applied to 54 Salmonella strains belonging to eight dominant serovars, and distinguished Infantis from the other seven serovars with an accuracy of 100%. The diagnostic primer set also did not show false positive amplification with 32 strains from eight non-Salmonella bacterial species. This cost-effective and rapid method can be considered an alternative to the classic serotyping using antisera.

scholarly journals Development of Quantitative Real-Time Polymerase Chain Reaction for Detection of and Discrimination between Erysipelothrix Rhusiopathiae and Other Erysipelothrix Species

2009 ◽  
Vol 21 (5) ◽  
pp. 701-706 ◽  
Author(s):  
Ho To ◽  
Tomohiro Koyama ◽  
Shinya Nagai ◽  
Kotaro Tuchiya ◽  
Tetsuo Nunoya
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Limit Of Detection ◽  
Enrichment Culture ◽  
Bacterial Species ◽  
Erysipelothrix Rhusiopathiae ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Practical Applications ◽  
Polymerase Chain

Quantitative real-time polymerase chain reaction (qPCR) assays were developed and validated in combination with enrichment culture for the detection and discrimination of Erysipelothrix rhusiopathiae and other Erysipelothrix species from tissue samples. The targets for SYBR green qPCR assays were the 16S ribosomal RNA gene for Erysipelothrix species and a gene involved in capsular formation for E. rhusiopathiae. The specificity of the assays was assessed with Erysipelothrix species and other related bacterial species. The limit of detection was found to be 5 colony-forming units per reaction. Amplification of DNA extracted from spleen and joint samples spiked with increasing quantities of Erysipelothrix cells was shown to be equally sensitive to DNA extracted from a pure bacterial culture. The assays were evaluated with 88 tissue samples from 3 experimentally infected pigs and 50 mice and with 36 tissue samples from 3 naturally infected pigs and 11 noninfected pigs. Results were compared with those of direct qPCR and conventional culture. The qPCR after enrichment increased the diagnostic sensitivity over that of culture and qPCR, thereby significantly reducing the total time taken for the detection of E. rhusiopathiae and other Erysipelothrix species. Therefore, this technique could be used for practical applications.

scholarly journals Identification of aflatoxigenic fungi using polymerase chain reaction-based assay

2014 ◽  
pp. 259-269 ◽  
Author(s):  
Vladislava Soso ◽  
Marija Skrinjar ◽  
Nevena Blagojev ◽  
Slavica Veskovic-Moracanin
Keyword(s):  
Polymerase Chain Reaction ◽  
Regulatory Gene ◽  
Control Point ◽  
Complex Mixture ◽  
Target Sequence ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Close Link ◽  
Critical Control Point ◽  
Polymerase Chain

As the aflatoxins represent a health-risk for humans because of their proven carcinogenicity, food-borne fungi that produce them as secondary metabolites, mainly Aspergillus flavus and Aspergillus parasiticus, have to be isolated and identified. The best argument for identifying problem fungi is that it indicates control points within the food system as part of a hazard analysis critical control point (HACCP) approach. This assumes there is a close link between fungus and toxin. Conventional methods for isolation and identification of fungi are time consuming and require admirably dedicated taxonomists. Hence, it is imperative to develop methodologies that are relatively rapid, highly specific and as an alternative to the existing methods. The polymerase chain reaction (PCR) facilitates the in vitro amplification of the target sequence. The main advantages of PCR is that organisms need not be cultured, at least not for a long time, prior to their detection, target DNA can be detected even in a complex mixture, no radioactive probes are required, it is rapid, sensitive and highly versatile. The gene afl-2 has been isolated and shown to regulate aflatoxin biosynthesis in A. flavus. Also, the PCR reaction was targeted against aflatoxin synthesis regulatory gene (aflR1) since these genes are nearly identical in A. flavus and A. parasiticus in order to indicate the possibility of detection of both the species with the same PCR system (primers/reaction). [Projekat Ministarstva nauke Republike Srbije, br. III46009] <br><br><font color="red"><b> This article has been retracted. Link to the retraction <u><a href="http://dx.doi.org/10.2298/APT1647265E">10.2298/APT1647265E</a><u></b></font>

scholarly journals Evaluation on the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats (Capra hircus) in Cebu, the Philippines

2019 ◽  
Vol 12 (6) ◽  
pp. 774-777 ◽  
Author(s):  
Adrian P. Ybañez ◽  
Orgil V. Arrabis ◽  
Dennis Justin M. Alvarez ◽  
Eloiza May S. Galon ◽  
Rhea Mae P. Jayag ◽  
...  
Keyword(s):  
Blood Smear ◽  
Peripheral Blood Smear ◽  
The Philippines ◽  
Capra Hircus ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Blood Smear Examination ◽  
Polymerase Chain ◽  
Babesia Spp

Background: Tick-borne diseases are caused by a wide variety of viruses, pathogens, and diseases. Anaplasma, Ehrlichia, and Babesia spp. are among the most known tick-borne pathogens in Asia. In the Philippines, these pathogens were already reportedly present in dogs and large ruminants, but no study has been reported yet evaluating their presence in goats. Aim: The present study aimed to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats in Cebu, the Philippines. Materials and Methods: A total of 100 blood samples from goats were collected in Cebu, the Philippines. Profile of sampled goats including age, body score, and sex was obtained. Peripheral blood smear examination and DNA extraction were performed. Nested polymerase chain reaction assay was used to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. Results: None of the samples were found positive with Anaplasma, Ehrlichia, and Babesia spp. infection. Conclusion: Tested goats were negative with Anaplasma, Ehrlichia, and Babesia spp. and calls for continuous surveillance of these pathogens due to the reported detection of these pathogens in other livestock animals in the area.

scholarly journals Isolation and identification of bacteria and parasites in glass eel (Anguilla spp.)

2021 ◽  
Vol 322 ◽  
pp. 02012
Author(s):  
Septyan Andriyanto ◽  
Hessy Novita ◽  
Tuti Sumiati ◽  
Taukhid
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Prevalence Rate ◽  
Bacterial Species ◽  
Bacterial Identification ◽  
Biochemical Method ◽  
Isolation And Identification ◽  
Glass Eel ◽  
Pathogenic Agent ◽  
Identification Of Bacteria ◽  
Glass Eels

The disease is the main agent that causes mortality of fish, especially during seed stages. The research aimed to find out bacteria and parasitic speciesin glass eel, Anguilla spp. Bacterial identification was carried out by a biochemical method. The prevalence of bacterial species was calculated using the El-Gohary et al. (2020) formula, while the results of bacterial identification from glass eel were Aeromonas spp., Vibrio spp., Enterococcus spp., Staphylococcus spp., Planococcus spp., Lactobacillus spp., Listeria spp., Citrbacterfreundii, Neisseria spp., Pseudomonas aeruginosa, Kurthia spp., Streptococcus spp., and Corynebacterium spp. It was found that the five highest prevalence rate was for Listeria spp. (39.64%), followed by Aeromonas spp. (26.13%), Staphylococcus spp. (16.22%), Corynebacterium spp. (5.41%), Lactobacillus spp. (2.70%), and the lowest prevalence rate was Streptococcus spp. (0.90%). The type of parasitic pathogen obtained was Trichodina spp. (2,70%), Dactylogyrus spp. (2,70%) and Gyrodactylus spp. (2,70%). Bacterial and parasites identified in glass eels need further verification on the epizootiology characteristic of each pathogenic agent.

scholarly journals Missed Plasmodium falciparum and Plasmodium vivax Mixed Infections in Ethiopia Threaten Malaria Elimination

2021 ◽  
Author(s):  
Colleen M. Leonard ◽  
Hussein Mohammed ◽  
Mekonnen Tadesse ◽  
Jessica N. McCaffery ◽  
Doug Nace ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodium Falciparum ◽  
Lactate Dehydrogenase ◽  
Plasmodium Vivax ◽  
Therapeutic Efficacy ◽  
Dried Blood Spots ◽  
Mixed Infections ◽  
Efficacy Study ◽  
Chain Reaction ◽  
Polymerase Chain

Plasmodium falciparum and Plasmodium vivax are co-endemic in Ethiopia. This study investigated whether mixed infections were missed by microscopy from a 2017 therapeutic efficacy study at two health facilities in Ethiopia. All patients (N = 304) were initially classified as having single-species P. falciparum (n = 148 samples) or P. vivax infections (n = 156). Dried blood spots were tested for Plasmodium antigens by bead-based multiplex assay for pan-Plasmodium aldolase, pan-Plasmodium lactate dehydrogenase, P. vivax lactate dehydrogenase, and histidine-rich protein 2. Of 304 blood samples, 13 (4.3%) contained both P. falciparum and P. vivax antigens and were analyzed by polymerase chain reaction for species-specific DNA. Of these 13 samples, five were confirmed by polymerase chain reaction for P. falciparum/P. vivax co-infection. One sample, initially classified as P. vivax by microscopy, was found to only have Plasmodium ovale DNA. Plasmodium falciparum/P. vivax mixed infections can be missed by microscopy even in the context of a therapeutic efficacy study with multiple trained readers.

Detection of Rhizobium meliloti cells in field soil and nodules by polymerase chain reaction

1995 ◽  
Vol 41 (9) ◽  
pp. 816-825 ◽  
Author(s):  
R. J. Watson ◽  
C. Haitas-Crockett ◽  
T. Martin ◽  
R. Heys
Keyword(s):  
Polymerase Chain Reaction ◽  
Rhizobium Meliloti ◽  
Genetically Engineered ◽  
Chain Reaction ◽  
Pcr Inhibitor ◽  
Vertical Location ◽  
Meliloti Strain ◽  
Identification Of Bacteria ◽  
Polymerase Chain ◽  
Genetically Engineered Microorganism

A genetically marked Rhizobium meliloti strain, R692, was prepared by insertion of a 1.7-kb DNA segment from Tn903 between the nifHDK and fixABC genes in the nod megaplasmid. This DNA was used as a marker, detectable by polymerase chain reaction (PCR), for the specific identification of bacteria in soil samples and alfalfa nodules. This detection technique was tested by applying different titres of the marked strain to field plots seeded with alfalfa. Samples of soil and nodules were assayed for the presence of the marker DNA fragment by PCR using primers specific to the marker sequence. The experiments revealed that the bacteria could be detected directly in soil containing about 103–104 bacteria/g, but greater sensitivity was prevented by potent PCR inhibitors present in the samples. The titre of the bacteria in the soil decreased rapidly after inoculation, dropping about 10-fold per week. Tests of vertical location of the bacteria in soil cores showed that the bacteria were initially dispersed to a depth of 18 cm, and subsequently retained viability in the top 2–8 cm. As few as 10 marked R. meliloti per gram of soil resulted in its establishment at detectable levels in nodules. Application of about 104–105 bacteria/g soil was sufficient to give the maximum number of nodules per plant and resulted in 70–90% occupancy by the marked strain. Limited movement of the inoculant was detected by analysis of nodules from plants adjacent to the sites where the bacteria were applied, probably by movement in water. The experiments demonstrated the advantages of PCR for the monitoring of marked microorganisms in the environment.Key words: genetically engineered microorganism, PCR inhibitor, nitrogen fixation, nif and fix genes, genetic marker.

Detection of Aspergillus fumigatus by Polymerase Chain Reaction (PCR)

2019 ◽  
Vol 10 (04) ◽  
pp. 655-661
Author(s):  
Zainab H Abood AL-Asadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Aspergillus Fumigatus ◽  
Oligonucleotide Primer ◽  
Rrna Gene ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain ◽  
The Impact

Aspergillosis refers to fungi infections of the respiratory tract caused by Aspergillus species, especially Aspergillus fumigatus. Infection of A. fumigatus was increased in the last few years due to either resistances to antibiotics or the influence of other factors such as other fungal infections. The present study aimed to review the impact of Aspergillus fumigatus in Aspergillosis cases, and study the role of Singleplex PCR for amplification of ITS1, ITS4 of rRNA gene in the detection of fungal isolate. In this study, One hundred sputum samples were collected from patients admitted to the specialize chest and respiratory diseases center / Baghdad who were suffering from respiratory problems. During these studied, molds were isolation and identification based on Conventional method (Direct microscopy by using 10% KOH, and fungal culture was done on Sabouraud Dextrose agar supplemented with chloramphenicol and on Czapek-Dox agar incubated at 37°C and examined for 3-7 days then macroscopic, microscopic examination of the colony by(lactophenol cotton blue stain )and molecular methods by using Polymerase chain reaction (PCR)technique for identification. The 10% KOH examination was positive for 35 cases, while laboratory culturing was positive for 53 cases. Aspergillus sp were isolated from 44(83%) patients; A. fumigatus was isolated in 23 (42. 4%) patients while A. flavus, A. niger, and A. terreus were isolated from 11 (20. 08%), (13. 2%) and 3 (5. 7%) patients respectively, also isolated Penicillium spp. at percentage 1(1. 9%). In this study. The ages of participants ranged from 10-70years with a mean age of 34years, the males were more susceptible to fungal infection, were recorded 35/53 (66. 3), compared to females were 18/53 (33. 96). The infection of fungi was more prevalent in ages 30-40recorded 26(53. 06%) followed by ages 40-50, 13(26. 5), while the lowest infection recorded in the age group 10- 20 years was 2(2. 04%). DNA isolated from twenty-three A. fumigatus isolates was used as a template, and the specific of oligonucleotide primer sequences were used in conventional PCR to detect the presence of internal transcribed spacer region ( ITS) region of the rRNA gene for Aspergillus fumigates. The results of the PCR amplification of the rRNA gene showed that this gene was present in 19 samples out 23 positive samples which isolation with a PCR product size of approximated 385 bp, while 4 samples out 23 positive samples showed negative results for the presence of this gene as indicated by the absence of the PCR products in their relevant lanes. Statistical analysis revealed that the PCR to have a sensitivity of 95. 1% in the detection of Aspergillus fumigatus in Aspergillosis cases. Polymerase chain reaction (PCR) is a rapid, specific, and sensitive method to detect Aspergillus fumigatus in aspergillosis cases of humans.

scholarly journals Development of PCR-Based Assays for Detecting Xanthomonas campestris pv. carotae, the Carrot Bacterial Leaf Blight Pathogen, from Different Substrates

Plant Disease ◽  
2004 ◽  
Vol 88 (11) ◽  
pp. 1226-1234 ◽  
Author(s):  
X. Q. Meng ◽  
K. C. Umesh ◽  
R. M. Davis ◽  
R. L. Gilbertson
Keyword(s):  
Xanthomonas Campestris ◽  
Random Amplified Polymorphic Dna ◽  
Bacterial Species ◽  
Leaf Blight ◽  
Bacterial Leaf Blight ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Agar Media ◽  
Extensive Collection ◽  
Polymerase Chain

Detection of the carrot bacterial leaf blight pathogen, Xanthomonas campestris pv. carotae, was achieved using polymerase chain reaction (PCR) along with primer pairs developed from sequences of cloned random amplified polymorphic DNA (RAPD) fragments. Primer pairs 3S and 9B directed the amplification of ∼350-bp and ∼900-bp (or ∼2 kb) DNA fragments, respectively, from genomic DNA of all known X. campestris pv. carotae strains tested, but not from that of 13 other X. campestris pathovars or other bacterial species, including yellow non-xanthomonad bacteria isolated from carrot tissues and seeds. In tests conducted with an extensive collection of X. campestris pv. carotae-like strains isolated from different substrates from California, Idaho, Oregon, Washington, and Canada, the 3S primer pair directed the amplification of the ∼350-bp target fragment from all strains. These results indicated that the 3S primer pair is highly specific for X. campestris pv. carotae detection. Using the 3S primer pair, PCR assays were developed for detection of X. campestris pv. carotae from colonies on agar media, carrot leaf and stem tissues, and seeds. These tests could be performed in a single day. The PCR-based seed assay detected X. campestris pv. carotae from lots with contamination rates ranging from 2 × 102 to 2.3 × 108 CFU per gram of seed. This assay gave results similar to a seed-wash dilution plating assay and proved more sensitive than an enzyme-linked immunosorbent assay (ELISA)-based assay.

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