scholarly journals Prevalence of the Pasteurellaceae bacteria in cattle farms of Moscow and Tver regions

2021 ◽  
Author(s):  
A.D. Kozlova ◽  
S.P. Yatsentyuk ◽  
D.A. Rudnyaev ◽  
Ju.I. Pobolelova
Keyword(s):  
Real Time Pcr ◽  
Respiratory Diseases ◽  
Pasteurella Multocida ◽  
Immune Status ◽  
Genetic Material ◽  
Animal Diseases ◽  
Bovine Respiratory Disease Complex ◽  
Etiological Agents ◽  
Bibersteinia Trehalosi ◽  
Cattle Farms

Pasteurellaceae bacteria are etiological agents of various animal diseases. Representatives of this family can be part of the "Bovine respiratory disease complex"" - a set of viral and bacterial pathogens that cause respiratory diseases in cattle. The aim of our work was to study the prevalence of Pasteurella multocida, Mannheimia haemolytica, Bibersteinia trehalosi, and Histophillus somni in calves aged 2 to 6 months in 37 farms in Moscow and Tver regions. Respiratory swabs were examined by Real time PCR. P. multocida DNA was found in all 37 farms (89% of samples). The genetic material of M. haemolytica and H. somni was detected less often - in 28% and in 18.5% of the samples. B. trehalosi DNA was detected in 3% of the samples from 8 farms. In one sample, P. multocida type F was detected, in 90.72% of positive P. multocida samples the presence of type A was confirmed. There was no circulation of P. multocida serotypes B, D, and E. Monoinfection with P. multocida was recorded only in 6 farms out of 37. In all other farms, a combination of two or more studied microorganisms was detected. An association of four pathogens was registered in four farms. The analysis of the frequency of P. multocida, M. haemolytica, B. trehalosi, and H. somni DNA detection in calves showed no correlation with the age of the animals. This may indicate that immune status and conditions of maintenance and care in the farm are of greatest importance in Pasteurellaceae prevalence in cattle.

DEVELOPMENT OF MULTIPLEX REAL-TIME PCR ASSAY FOR SIX PATHOGENS: RAPID TOOL FOR DIAGNOSIS OF BACTERIAL MENINGITIS

2019 ◽  
pp. 60-66
Author(s):  
Viet Quynh Tram Ngo ◽  
Thi Ti Na Nguyen ◽  
Hoang Bach Nguyen ◽  
Thi Tuyet Ngoc Tran ◽  
Thi Nam Lien Nguyen ◽  
...  
Keyword(s):  
Bacterial Meningitis ◽  
Real Time ◽  
Real Time Pcr ◽  
Diagnostic Methods ◽  
Type B ◽  
E Coli ◽  
Pcr Assays ◽  
Set Up ◽  
Etiological Agents ◽  
Suis Serotype

Introduction: Bacterial meningitis is an acute central nervous infection with high mortality or permanent neurological sequelae if remained undiagnosed. However, traditional diagnostic methods for bacterial meningitis pose challenge in prompt and precise identification of causative agents. Aims: The present study will therefore aim to set up in-house PCR assays for diagnosis of six pathogens causing the disease including H. influenzae type b, S. pneumoniae, N. meningitidis, S. suis serotype 2, E. coli and S. aureus. Methods: inhouse PCR assays for detecting six above-mentioned bacteria were optimized after specific pairs of primers and probes collected from the reliable literature resources and then were performed for cerebrospinal fluid (CSF) samples from patients with suspected meningitis in Hue Hospitals. Results: The set of four PCR assays was developed including a multiplex real-time PCR for S. suis serotype 2, H. influenzae type b and N. meningitides; three monoplex real-time PCRs for E. coli, S. aureus and S. pneumoniae. Application of the in-house PCRs for 116 CSF samples, the results indicated that 48 (39.7%) cases were positive with S. suis serotype 2; one case was positive with H. influenzae type b; 4 cases were positive with E. coli; pneumococcal meningitis were 19 (16.4%) cases, meningitis with S. aureus and N. meningitidis were not observed in any CSF samples in this study. Conclusion: our in-house real-time PCR assays are rapid, sensitive and specific tools for routine diagnosis to detect six mentioned above meningitis etiological agents. Key words: Bacterial meningitis, etiological agents, multiplex real-time PCR

scholarly journals Estimating Clinically Relevant Cut-Off Values for a High-Throughput Quantitative Real-Time PCR Detecting Bacterial Respiratory Pathogens in Cattle

2021 ◽  
Vol 8 ◽  
Author(s):  
Alicia F. Klompmaker ◽  
Maria Brydensholt ◽  
Anne Marie Michelsen ◽  
Matthew J. Denwood ◽  
Carsten T. Kirkeby ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Age Groups ◽  
Therapeutic Interventions ◽  
Respiratory Pathogens ◽  
Logistic Regression Models ◽  
Histophilus Somni ◽  
Nasal Swabs

Bovine respiratory disease (BRD) results from interactions between pathogens, environmental stressors, and host factors. Obtaining a diagnosis of the causal pathogens is challenging but the use of high-throughput real-time PCR (rtPCR) may help target preventive and therapeutic interventions. The aim of this study was to improve the interpretation of rtPCR results by analysing their associations with clinical observations. The objective was to develop and illustrate a field-data driven statistical method to guide the selection of relevant quantification cycle cut-off values for pathogens associated with BRD for the high-throughput rtPCR system “Fluidigm BioMark HD” based on nasal swabs from calves. We used data from 36 herds enrolled in a Danish field study where 340 calves within pre-determined age-groups were subject to clinical examination and nasal swabs up to four times. The samples were analysed with the rtPCR system. Each of the 1,025 observation units were classified as sick with BRD or healthy, based on clinical scores. The optimal rtPCR results to predict BRD were investigated for Pasteurella multocida, Mycoplasma bovis, Histophilus somni, Mannheimia haemolytica, and Trueperella pyogenes by interpreting scatterplots and results of mixed effects logistic regression models. The clinically relevant rtPCR cut-off suggested for P. multocida and M. bovis was ≤ 21.3. For H. somni it was ≤ 17.4, while no cut-off could be determined for M. haemolytica and T. pyogenes. The demonstrated approach can provide objective support in the choice of clinically relevant cut-offs. However, for robust performance of the regression model sufficient amounts of suitable data are required.

scholarly journals Diagnostics of main bacterial agents of porcine respiratory diseases complex (PRDC) using PCR detection of Mycoplasma hyopneumoniae

2012 ◽  
Vol 49 (No. 2) ◽  
pp. 35-41 ◽  
Author(s):  
I. Holko ◽  
J. Urbanova ◽  
THolkova ◽  
V. Kmet
Keyword(s):  
Respiratory Diseases ◽  
Pasteurella Multocida ◽  
Respiratory Infections ◽  
Clinical Signs ◽  
Antibiotic Sensitivity ◽  
Mycoplasma Hyopneumoniae ◽  
Sensitivity Testing ◽  
Pcr Method ◽  
One Year ◽  
Porcine Respiratory

The main goal of our work is the presentation and analysis of incidence of porcine respiratory disease complex (PRDC) regarding bacterial agents in the territory of northern districts of Slovakia. Mycoplasma hyopneumoniae and other secondary bacterial causative pathogens of PRDC comprised 75.2% of all cases (98) with clinical signs of respiratory infections that we examined in the course of one year. We present also one of possibilities to the solution of problematic detection of M. hyopneumoniae which is, like the whole rank of mycoplasmas, very difficult to cultivate. This problem was solved by using the PCR method with the direct isolation of M. hyopneumoniae from lungs tissue. In antibiotic sensitivity testing of Pasteurella multocida and Actinobacillus pleuropneumoniae resulted enrofloxacin as the most effective antibiotics in the therapy of PRDC regarding bacterial agents.in above mentioned territory.

scholarly journals DNA recovery from archived RDTs for genetic characterization of Plasmodium falciparum in a routine setting in Lambaréné, Gabon

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
The Trong Nguyen ◽  
Brice Nzigou Mombo ◽  
Albert Lalremruata ◽  
Erik Koehne ◽  
Rella Zoleko Manego ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Real Time ◽  
Real Time Pcr ◽  
Rural Areas ◽  
Large Scale ◽  
Genetic Material ◽  
Chloroquine Resistance ◽  
Dna Recovery ◽  
Parasite Populations ◽  
Pcr Targeting

Abstract Background Rapid diagnostic tests (RDTs) have been described as a source of genetic material to analyse malaria parasites in proof-of-concept studies. The increasing use of RDTs (e.g., in focal or mass screening and treatment campaigns) makes this approach particularly attractive for large-scale investigations of parasite populations. In this study, the complexity of Plasmodium falciparum infections, parasite load and chloroquine resistance transporter gene mutations were investigated in DNA samples extracted from positive RDTs, obtained in a routine setting and archived at ambient temperature. Methods A total of 669 archived RDTs collected from malaria cases in urban, semi-urban and rural areas of central Gabon were used for P. falciparum DNA extraction. Performance of RDTs as a source of DNA for PCR was determined using: (i) amplification of a single copy merozoite surface protein 1 (msp1) gene followed by highly sensitive and automated capillary electrophoresis; (ii) genotyping of the pfcrt gene locus 72–76 using haplotype-specific-probe-based real-time PCR to characterize chloroquine resistance; and, (iii) real-time PCR targeting 18S genes to detect and quantify Plasmodium parasites. Results Out of the 669 archived RDTs, amplification of P. falciparum nucleic materials had a success rate of 97% for 18S real-time PCR, and 88% for the msp1 gene. The multiplicity of infections (MOI) of the whole population was 2.6 (95% CI 2.5–2.8). The highest number of alleles detected in one infection was 11. The MOI decreased with increasing age (β = − 0.0046, p = 0.02) and residence in Lambaréné was associated with smaller MOIs (p < 0.001). The overall prevalence of mutations associated with chloroquine resistance was 78.5% and was not associated with age. In Lambaréné, prevalence of chloroquine resistance was lower compared to rural Moyen-Ogooué (β = − 0.809, p-value = 0.011). Conclusion RDT is a reliable source of DNA for P. falciparum detection and genotyping assays. Furthermore, the increasing use of RDTs allows them to be an alternative source of DNA for large-scale genetic epidemiological studies. Parasite populations in the study area are highly diverse and prevalence of chloroquine-resistant P. falciparum remains high, especially in rural areas.

scholarly journals RETROSPECTIVE EPIDEMIOLOGIC STUDY OF DISEASES IN RUMINANTS IN KHAGRACHARI HILL TRACT DISTRICT OF BANGLADESH

2013 ◽  
Vol 9 (2) ◽  
pp. 145-153 ◽  
Author(s):  
MH Ali ◽  
MKJ Bhuiyan ◽  
MM Alam
Keyword(s):  
Respiratory Diseases ◽  
Skin Diseases ◽  
Epidemiologic Study ◽  
Gastrointestinal Diseases ◽  
Lower Percentage ◽  
Age Group ◽  
Animal Diseases ◽  
Worm Infestation ◽  
High Prevalence ◽  
Veterinary Hospital

A retrospective epidemiologic study of animal diseases was undertaken at Khagrachari Sadar Veterinary Hospital during January, 2006 to December, 2010 to determine prevalence and distribution of animal diseases. According to the diseases register, a total of 3988 sick animals were examined and 53 types of diseases were identified during this period. The commonly found various diseases were worm infestation (51.5%), pneumonia and pneumonitis (7.9%), ephemeral fever (3.7%), enteritis (3.4%), mastitis(3.2%), mange (3.2%), indigestion (2.8%), anestrous(2.6%). Rest of the diseases had lower percentage than 2%. Out of 3988 sick animals, 74.7% were female and 25.3 % were male animals. Animals aged between 2-5 (A1) years had high prevalence (54.0%) and it was low in age group 8-10 years (A4), 2.4%. Prevalence of diseases was high (42.3%) in rainy season (June-October) followed by (32.5%) in winter (November-February) and lowest (25.2%) in summer season (March-May). Gastrointestinal diseases 61.6 % (2458 cases) was seen highly prevalent among all groups of animals which was followed by infectious diseases 10.4% (416 cases), skin diseases 9.4 % (377 cases), respiratory diseases 8.27% (330 cases) and reproductive diseases 7.93% (cases). This study suggests that for a period of 15 years or more will help to identify the risk factors of diseases in this area.DOI: http://dx.doi.org/10.3329/bjvm.v9i2.13457

scholarly journals Environmental Attributes to Respiratory Diseases of Small Ruminants

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Anu Rahal ◽  
Abul Hasan Ahmad ◽  
Atul Prakash ◽  
Rajesh Mandil ◽  
Aruna T. Kumar
Keyword(s):  
Disease Burden ◽  
Respiratory Diseases ◽  
Immune Status ◽  
Respiratory Health ◽  
Environmental Pollutants ◽  
Small Ruminants ◽  
Parasitic Infections ◽  
Environmental Attributes ◽  
New Compounds ◽  
Alveolar Surface

Respiratory diseases are the major disease crisis in small ruminants. A number of pathogenic microorganisms have been implicated in the development of respiratory disease but the importance of environmental factors in the initiation and progress of disease can never be overemphasized. They irritate the respiratory tree producing stress in the microenvironment causing a decline in the immune status of the small ruminants and thereby assisting bacterial, viral, and parasitic infections to break down the tissue defense barriers. Environmental pollutants cause acute or chronic reactions as they deposit on the alveolar surface which are characterized by inflammation or fibrosis and the formation of transitory or persistent tissue manifestation. Some of the effects of exposures may be immediate, whereas others may not be evident for many decades. Although the disease development can be portrayed as three sets of two-way communications (pathogen-environment, host-environment, and host-pathogen), the interactions are highly variable. Moreover, the environmental scenario is never static; new compounds are introduced daily making a precise evaluation of the disease burden almost impossible. The present review presents a detailed overview of these interactions and the ultimate effect on the respiratory health of sheep and goat.

scholarly journals The efficiency of sodC gene / N. meningitidis detection in comparison with the classical methods for the diagnosis of meningococcal infection / Evaluarea eficienţei Real Time PCR TaqMan utilizând gena sodC / N. meningitidis în comparaţie cu metodele clasice utilizate în diagnosticul infecţiei meningococice

2015 ◽  
Vol 23 (1) ◽  
Author(s):  
Roxana Elena Nemescu ◽  
Ramona Gabriela Ursu ◽  
Carmen Mihaela Dorobăț ◽  
Luminița Smaranda Iancu
Keyword(s):  
Antibiotic Therapy ◽  
Real Time ◽  
Real Time Pcr ◽  
Latex Agglutination ◽  
Blood Analysis ◽  
Meningococcal Infection ◽  
Specific Method ◽  
Rt Pcr ◽  
Highly Sensitive ◽  
Etiological Agents

AbstractMeningococcal infection requires a fast and accurate diagnostic method in order to correctly initiate the antibiotic therapy. The aim of our study was to assess the efficiency of Real Time PCR -Taq Man using sod C gene / N. meningitidis in comparison with the classical methods for the diagnosis of meningococcal infection - direct microscopy, cultivation, latex agglutination and blood culture. We have detected 24/44 (54.54%) patients with meningococcal infection. In both cases of patients with / without previous antibiotic therapy before admission, the AUC (area under curve) had the highest values for RT PCR in CSF and blood analysis. This sod C RT-PCR assay is a highly sensitive and specific method for detection of Neisseria meningitis and it would be useful to include this method like a multiplex in routine testing of patients with clinical meningococcal infection for other etiological agents also.

scholarly journals Etiological characteristics of acute respiratory diseases in servicemen of the Western military district in 2014-2019

2020 ◽  
Vol 22 (1) ◽  
pp. 81-86
Author(s):  
A N Gorenchuk ◽  
P V Kulikov ◽  
S D Zhogolev ◽  
R M Aminev ◽  
A A Kuzin ◽  
...  
Keyword(s):  
Environmental Sustainability ◽  
Respiratory Diseases ◽  
Respiratory Infections ◽  
Genetic Material ◽  
Influenza Viruses ◽  
Respiratory Pathogens ◽  
Acute Respiratory Diseases ◽  
Socio Demographic Factors ◽  
Comparison Of The Results ◽  
Active Implementation

The species affiliation of respiratory pathogens isolated from patients and carriers in the military units of the Western Military District in 2014-2019 was studied. The analysis of long-term and seasonal dynamics of their circulation is carried out. It was found that S. pneumoniae and adenoviruses are more often detected in acute respiratory diseases in conscripts. The genetic material of adenoviruses was found in 31,9% of samples, influenza viruses in 13,3%, rhinoviruses in 11,2%, respiratory syncytial viruses in 1,7%, metapneumoviruses in 0,9%, parainfluenza viruses 0,7%, bocaviruses0,5%, coronaviruses 0,1%, S. pneumoniae 33,9%, H. influenzae 13%, M. pneumoniae 9%, C. pneumoniae - in 3,3%, N. meningitidis - in 16%. Comparison of the results of work with studies carried out by domestic research groups among the civilian population in the same period showed that the circulation of various respiratory viruses depends on the year, season, and is also influenced by socio-demographic factors. A direct high functional correlation was found between the dynamics of circulation of adenovirus and S. pneumoniae in different years and epidemic seasons. Evidence has been obtained of the active implementation of the process of self-maintenance of the reservoir of infections and the multifactorial nature of the overall environmental sustainability of the system in organized military teams. In the etiological structure of respiratory infections, the proportion of pathogens varies depending on the season in different years, the characteristics of the formation and composition of organized groups, as well as epidemic periods.

scholarly journals Detection of the BCR-ABL Gene By the Real-Time PCR Method in Patients with Chronic Myeloid Leukemia in Rio Grande Do Norte, Brazil

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5432-5432
Author(s):  
Aldair Sousa Paiva ◽  
Hugo Diogenes De Oliveira Paiva ◽  
Geraldo Barroso Cavalcanti ◽  
Gioconda DR Leão ◽  
Marcos Dias Leão ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Real Time ◽  
Real Time Pcr ◽  
Myeloid Leukemia ◽  
Conflicts Of Interest ◽  
Rna Extraction ◽  
Philadelphia Chromosome ◽  
Genetic Material ◽  
Chromosome 9 ◽  
The Real

Abstract Background: The Philadelphia chromosome is a cytogenetic change resulting from a reciprocal translocation of genetic material between ABL genes from chromosome 9 and BCR from chromosome 22 or t(9; 22) (q34; 11), forming the chimeric gene BCR- ABL, being associated with chronic myeloid leukemia (CML), acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML). The p190 variant is usually associated with acute forms of leukemia, including AML and ALL, whereas the p210 variant is associated with the chronic phases of CML. Due to the high sensitivity and specificity, nucleic acid amplification techniques by real-time PCR have replaced the conventional cytogenetic techniques for the identification of the Philadelphia chromosome and its p190 and p210 variants. Molecular analysis has been indicated in the initial diagnostic phase and also for the therapeutic monitoring defining the percentage of neoplastic cells present in the patients during the different phases of the treatment (Minimum Residual Disease or MRD).The aim of this study was the transcript BCR-ABL identification in patients with suspected of CML and evaluation of the gene frequency in these patients. Methods: The presence of BCR-ABL gene was investigated in blood samples from 42 patients with suspected CML. The RNA extraction was performed by phenol/chloroform method. The cDNA was submitted to PCR, using specific primers for and BCR-ABL genes by Real time PCR. Results: From all studied patients, 16 (38.10%) were negative, and 26 (59.09%) positive for one of rearrangements: p210 b3a2 and b2a2 in 18 cases (40.91%) and p190 a1a2 in 2 cases (4,76%) and double positive p120/190 in 6 cases (14,28%). We observed that the most common rearrangement was the p210 b3a2, and the molecular results were compatible with clinical and hematologic suspicion. Conclusions: The Real-timePCR, because of its specificity and sensitivity, can be considered the most used technique in routine diagnosis and investigation of MRD of CML patients. Disclosures No relevant conflicts of interest to declare.

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