scholarly journals Pathology of Fowl Paratyphoid and Molecular Detection of Its Pathogen in Layer Chickens

2021 ◽  
Vol 24 (2) ◽  
pp. 47-57
Author(s):  
J Alam ◽  
T Chakma ◽  
MS Islam ◽  
MT Islam ◽  
MAHNA ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Salmonella Enteritidis ◽  
Molecular Detection ◽  
Inflammatory Cells ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Salmonella Spp ◽  
Gross Lesions ◽  
Pcr Method ◽  
Polymerase Chain

The study was aimed to ascertain the pathology of fowl paratyphoid and molecular detection of its causal agent (Salmonella spp) in chickens. Pathological and swab samples were collected from layers in Gazipur district, Bangladesh. For observing the gross and microscopic lesions of different organs necropsy and histopathology were done, and to isolate and identify the Salmonella spp, different bacteriological tests and Polymerase Chain Reaction (PCR) were performed. Swabs from 150 chickens showed 66% of salmonellosis. Gram’s staining of isolated bacteria showed pink colored rod shaped bacilli. In biochemical tests, Salmonella fermented dextrose, maltose, xylose, arabinose, dulcitol, mannitol except lactose and sucrose. Investigation of gross lesions at necropsy revealed hemorrhage and congestion in intestine, liver, spleen and ovaries. Necrotic foci were found in liver and spleen, and button like ulceration in cecal tonsils as well. Microscopic lesions included hemorrhage and focal necrosis in liver and spleen. Congestion and infiltrations of inflammatory cells were observed in small intestine. Ovary was hemorrhagic and there was infiltration of heterophils. Biochemically positive and isolated Salmonella organisms were confirmed by PCR method using invA and IE1 primers. The final results showed that a total of 91.7% Salmonella suspected cultures were confirmed as Salmonella Enteritidis. Ann. Bangladesh Agric. (2020) 24(2) : 47-57

Combination of Immunomagnetic Separation and Polymerase Chain Reaction for the Simultaneous Detection of Listeria monocytogenes and Salmonella spp. in Food Samples

2001 ◽  
Vol 64 (11) ◽  
pp. 1744-1750 ◽  
Author(s):  
HSIEN-YEE HSIH ◽  
HAU-YANG TSEN
Keyword(s):  
Polymerase Chain Reaction ◽  
Listeria Monocytogenes ◽  
Simultaneous Detection ◽  
Meat Products ◽  
Immunomagnetic Separation ◽  
Food Samples ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Pcr Method ◽  
Polymerase Chain

A method that combined the immunomagnetic separation (IMS) technique and the multiplex polymerase chain reaction (PCR) method (i.e., the IMS-mPCR method) was developed for simultaneous detection of Listeria monocytogenes and Salmonella spp. in food samples. When only the multiplex PCR method was used, it was found that if cell numbers of each of the two target organisms (L. monocytogenes and Salmonella spp.) were above the detection limit, but differed by more than 2 logs—e.g., n × 107 to n × 104 or n × 106 to n × 103—the organism presenting the lower numbers might go undetected. Following the enrichment step with universal preenrichment (UP) broth, if an IMS method using equal quantities of anti-Listeria and anti-Salmonella immunomagnetic beads was performed prior to PCR, both pathogens could be detected unambiguously. Such results could be obtained for target organisms in food samples, such as milk, dairy, and meat products, if similar enrichment and IMS steps were performed prior to PCR.

scholarly journals Molecular Detection of Plasmid-Mediated Quinolone Resistance in Ciprofloxacin-Resistant Escherichia coli from Urine of Patients attending Garki Hospital, Abuja, Nigeria

2020 ◽  
Vol 1 (4) ◽  
Author(s):  
Moses Oghenaigah Eghieye ◽  
Istifanus Haruna Nkene ◽  
Rejoice Helma Abimiku ◽  
Yakubu Boyi Ngwai ◽  
Ibrahim Yahaya ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Molecular Detection ◽  
Quinolone Resistance ◽  
Chain Reaction ◽  
E Coli ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections

Urinary tract infections (UTIs) caused by Escherichia coli (E. coli) is common worldwide; and its successful treatment using antibiotics is limited by acquisition of resistance by the bacteria. This study investigated the occurrence of plasmid-mediated quinolone resistance (PMQR) genes in ciprofloxacin-resistant E. coli from urine of patients with suspected cases of UTIs attending Garki Hospital Abuja (GHA), Nigeria. A total of 8 confirmed ciprofloxacin-resistant E. coli was screened for carriage of PMQR genes using polymerase chain reaction (PCR) method. The occurrences of the PMQR genes detected were in the order: aac-(6′)-Ib-cr (87.5%) > qnrB (50.0%) > qnrS (37.5%) > oqxAB (12.5%) > qnrA(0.0%). qnrB and qnrS did not exist alone, but in combination with other genes; aac-(6′)-Ib-crexisted both alone and in combination with others; the most prevalent patterns of existence were aac-(6′)-Ib-cr alone and aac-(6′)-Ib-cr + qnrB + qnrS at 25.0% each. This study has shown that the ciprofloxacin-resistant E. coli harbored aac-(6′)-Ib-cr, qnrB, qnrS and oqxAB PMQR genes, with aac-(6′)-Ib-cr being the most prevalent. The genes were present either alone or in combination with one another. This has implication for the clinical application of fluoroquinolones to treat UTI in the study location and environs. 

scholarly journals Detection Method for Salmonella Typhimurium and Salmonella Enteritidis using Real-Time Polymerase Chain Reaction

2019 ◽  
Vol 7 (4.14) ◽  
pp. 302 ◽  
Author(s):  
Siti Nurjanah ◽  
Winiati P. Rahayu ◽  
Lisa Al Mutaqin
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Salmonella Enteritidis ◽  
Shigella Dysenteriae ◽  
Virulence Plasmid ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Specific Target Gene ◽  
Polymerase Chain ◽  
Primer Sequence

Salmonella is a pathogenic bacterium that can cause serious harm to humans. Chicken carcasses have been reported contaminated by Salmonella, especially S. Typhimurium and S. Enteritidis. These two serovars are very difficult to be confirmed and distinguished using biochemical analysis, therefore a rapid method for detection and differentiation of both is required. The objective of this study was to evaluate designed primer for detection and differentiation of S. Typhimurium and S. Enteritidis on chicken carcasses using real time Polymerase Chain Reaction (rt-PCR). Detection of Salmonella spp. was conducted using primer sequence from invA gene. Differentiation of both Salmonella serovars was conducted using specific target gene from S. Typhimurium (STM) and specific virulence plasmid of S. Enteritidis (Prot6E). The result showed that invA primer effective to detect all species Salmonella tested and has good specificity that could not detect Escherichia coli and Shigella dysenteriae in the similar melting temperature.   Two specific primers STM and prot6E have distinguished between S. Typhimurium and S. Enteritidis.  Sensitivity of method showed very good with 0.5 μM primer concentration of invA, STM and prot6E that were 0.2 pg, 22 pg and 28 pg respectively. Initial trial showed that this method can be applied for detection of Salmonella spp. and two serovars in chicken carcasses.  

scholarly journals Screening of Salmonella spp. and Chlamydophila psittaciin parrots domiciled in Rio Branco, Acre, Brazil

2021 ◽  
Vol 15 (4) ◽  
pp. 339-344
Author(s):  
Breno Kalyl Freitas Nascimento ◽  
Luciana dos Santos Medeiros ◽  
Leandro dos Santos Machado ◽  
Vânia Maria França Ribeiro
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Infectious Diseases ◽  
Emerging Infectious Diseases ◽  
Wild Animals ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Salmonella Spp ◽  
Zoonotic Pathogens ◽  
Polymerase Chain

A large proportion of emerging infectious diseases (60.3%) globally are zoonotic pathogens, and of these, 71.8% originate from wild animals. Salmonellosis and psittacosis, diseases caused by Salmonella spp. and Chlamydophila psittaci, respectively, in wild animals are zoonoses with great risks to public health. Therefore, this study aimed to investigate the presence of Salmonella spp. and C. psittaci in parrots domiciled in Rio Branco, Acre. The animals in the study were raised as pets, and selection was performed based on convenience criteria. The birds were manually restrained to collect biological materials. Subsequently, conventional microbiological and biochemical tests were performed to identify Salmonella spp., and polymerase chain reaction analyses were conducted to identify C. psittaci and Salmonella spp. It was not possible to isolate Salmonella spp. and C. psittaci in the sampled birds. However, the presence of these bacteria in parrots cannot be ruled out because intermittent release and diagnostic limitations are widely described in the literature.

scholarly journals DETECTION OF SALMONELLA SPP. IN COMMERCIAL EGGS FROM LAYER CHICKEN FARMS AND TRADITIONAL MARKETS IN BALI

2021 ◽  
Vol 22 (1) ◽  
pp. 93-100
Author(s):  
Alifianita Anake Yansri ◽  
Hani Plumeriastuti ◽  
Mustofa Helmi Effendi
Keyword(s):  
Polymerase Chain Reaction ◽  
Salmonella Enteritidis ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Traditional Markets ◽  
Layer Chicken ◽  
Commercial Chicken ◽  
Polymerase Chain ◽  
Duplex Polymerase Chain Reaction ◽  
Chicken Eggs

This study aims to early detect Salmonella spp. and identify serotypes in commercial chicken eggs from layer chicken farms and traditional markets in Bali. Early detection study of Salmonella spp. was carried out by conventional bacteriological methods, while serotype identification by duplex Polymerase Chain Reaction (d-PCR) test against the invA gene from Salmonella spp. and the sefA gene from Salmonella enteritidis. Egg samples in this study were taken from 10 layer chicken farms in Bali which included districts of Bangli, Gianyar, Tabanan and Karangasem. Egg samples from traditional markets were taken from 18 traditional markets from the districts of Bangli, Gianyar, Tabanan, Karangasem, Badung, and Denpasar City. Samples were eggshells and egg whites. Analysis of positive results from Salmonella spp. described descriptively. The results showed that eggshells and white eggs from all of the layer chicken farms are negative contaminated with Salmonella spp. (0%). In eggshell samples taken from the traditional markets of Taman Bali and Tulikup from the districts of Bangli and Gianyar, positive with Salmonella spp. (11,1%) by conventional bacteriological tests. In the duplex Polymerase Chain Reaction test, S. enteritidis serotypes were identified. The finding contamination of Salmonella enteritidis in commercial chicken eggs from traditional markets require periodically detection to prevent the occurrence of salmonellosis due to consumption of contaminated chicken eggs in traditional markets in Bali.

scholarly journals Detection of Aedes aegypti Resistance towards Cypermethrin using Polymerase Chain Reaction (PCR) Techniques in Ambarawa Semarang Regency

2020 ◽  
Vol 9 (1) ◽  
pp. 71-77
Author(s):  
Widya Hary Cahyati ◽  
Laila Fitriani
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Molecular Detection ◽  
Sampling Techniques ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Homozygous Resistant ◽  
Development Center ◽  
Pcr Techniques

Ambarawa Regency is one of the endemic sub-districts since 2017 which always accounts for the highest DHF cases in Semarang Regency. Molecular detection of Aedes aegypti resistance towards sipermetrin using Polymerase Chain Reaction (PCR) techniques. The study was conducted to see mutations in the Aedes aegypti VGSC gene. This research was a pure experimental study. The Aedes aegypti samples examined were 10 in every village taken from 3 endemic DHF villages with high fogging intensity in Ambarawa. The research used random sampling techniques taken with ovitrap which was first installed during the month of August. Resistance detection test using the PCR method were conducted at the Banjarnegara Research and Development Center in September. Data were analyzed descriptively to illustrate the results of the study. The results showed that in Tambakboyo Village, 2 samples were susceptible (V / V), 7 samples were detected as homozygous (G / G) resistant, 1 sample was detected as heterozygous (V / G) resistant; in Kupang District 5 samples were detected as being homozygous resistant (G / G) and 5 samples were detected as heterozygous resistant (V / G); and in Panjang Village 1 susceptible sample (V / V), 8 samples were detected as homozygous resistant (G / G), 1 sample was detected as heterozygous resistant (V / G). Based on the results of mutation studies had been found in the VGSC gene in codon V1016G. The proper implementation of management, selection and rotation of insecticides is expected to reduce the risk of resistance in the Aedes aegypti mosquito population.

scholarly journals Conventional and molecular detection of vibrio cholerae isolated from environmental water with the prevalence of antibiotic resistance mechanisms.

2019 ◽  
Vol 10 (3) ◽  
Author(s):  
Suad A Al-Hilu ◽  
Ali M Al-Mohana ◽  
Zainab Jaber
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Molecular Detection ◽  
Public Health Problem ◽  
Environmental Water ◽  
Selective Medium ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Vibrio Cholera ◽  
Polymerase Chain

Environmental water is an important source for Vibrio cholerae, which is autochthonous to the aquatic environment, monitoring this bacterium in water is important for control of cholera. Vibrio cholerae represents an enormous public health problem around the world, especially in developing countries. One hundred samples were collected and selected. The samples were filtered and transferred to slants containing 2ml of alkaline peptone water, then subcultured on selective medium Thiosulphate Citrate Bile Salt Sucrose agar. All presumptive isolates were confirmed by using a series of biochemical tests including Oxidase test, Simmon Citrate test, DNase test, Indole test, Klingler Iron Agar (KIA) test, MacConkey agar test and motility. Confirmed Vibrio cholera strains were then screening for slide agglutination test by using commercially antisera polyvalent and monovalent O1 and O139 for determining strain serotype. The DNA extracted from pure culture and Polymerase Chain Reaction assay was used for molecular detection of Vibrio cholerae, a specific primer which designed according to ctxA gene sequences. This primer was detection and amplifying 241 base pairs of the ctxA gene. The resistance to antibiotics by Vibrio cholerae was determining by using thirteen standardized disc diffusion including Amikacin, Ceftriaxone, Ceftazidime, Gentamycin, Tetracycline, Streptomycin, Tobramycin, Cephotaxime, Nalidixic Acid, Norfloxacin, Cephalothin, Rifampicin, Cefixime. From one hundred water samples were detected, fifty-six samples were motile and positive for biochemical tests. Fifteen isolates confirmed as Vibrio cholera by Polymerase Chain Reaction (PCR) assay with primers de­signed for ctxA and 241bp band was observed. These fifteen isolates showed agglutination with polyvalent and monovalent O1 antisera, and two strains represented Ogawa from other strains that showed Inaba. The fifteen isolates exhibited an identical response to each antibiotic examined. They showed sensitive to all antibiotics except Amikacin, Streptomycin, Cefixime, Norfloxacin, Cephalothin. the aim of this study was determined the accurate method for detection of Vibrio cholerae in environmental water. In the current study, we found that the molecular method using Polymerase Chain Reaction performance using the ctxA gene-specific primers for detection of Vibrio cholerae was faster and accurate and specific.

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