scholarly journals Molecular Detection and Characterization of Pasteurella multocida Infecting Camels in Marsabit and Turkana Counties, Kenya

Author(s):  
Justus Kyalo Kasivalu ◽  
George Isanda Omwenga ◽  
Gabriel Oluga Aboge

Abstract BackgroundInfection with Pasteurella multocida is abundant in Kenya yet there is scarce information on their genetic diversity. Pasteurella multocida is considered to be one of the normal flora in the respiratory tract of camels and other animals but it becomes pathogenic and causes pasteurellosis when the resistance of the camel body is diminished by harmful environmental influences. Close herding, overwork, limited food supply, and wet climatic conditions are stresses that seem to speed the spread of the infection. Conventional PCR, Multiplex PCR and sequencing were applied to enhance identification of Pasteurella multocida at any level of specificity viz; strain, species, and genus. These molecular tools were applied to confirm the presence and genetic diversity of Pasteurella multocida in 102 blood and 30 nasal swab samples collected from Marsabit and Turkana counties in Kenya. Kmt1 gene was used as the marker gene for Pasteurella multocida and hyaD-hyaC, bcbD, dcbF, ecbJ, and fcbD as marker genes for capsular groups. A study done in northern Kenya noted that in Africa pasteurellosis infections causing death in camels (Camelus dromedarius) have been existing since 1890 though the real cause of this disease remains elusive and needs further study. The study was done to detect Pasteurella multocida and characterize its capsular types by application of molecular biology toolsResultsTwenty one Kenyan isolates were confirmed to be Pasteurella multocida and only capsular group E was detected in both counties. Pasteurella multocida sequences were found to be highly conserved, however isolates detected in Kenya were found to be genetically related to other isolates from African and other parts of the world. ConclusionsThe study confirm that the camels were infected by Pasteurella multocida of capsular type E in Marsabit and Turkana Counties of Kenya. DNA sequences were found to be homologous to Pasteurella multocida thereby confirming that the camels were infected by Pasteurella multocida.

2003 ◽  
Vol 93 (5) ◽  
pp. 596-603 ◽  
Author(s):  
Jeri D. Barak ◽  
Robert L. Gilbertson

Bacterial leafspot of lettuce (BLS), caused by Xanthomonas campes-tris pv. vitians, has become more prevalent in many lettuce-growing areas of the world over the past decade. To gain insight into the nature of these outbreaks, the genetic variation in X. campestris pv. vitians strains from different geographical locations was examined. All strains were first tested for pathogenicity on lettuce plants, and then genetic diversity was assessed using (i) gas-chromatographic analysis of bacterial fatty acids, (ii) polymerase chain reaction analysis of repetitive DNA sequences (rep-PCR), (iii) DNA sequence analysis of the internal transcribed spacer region 1 (ITS1) of the ribosomal RNA, (iv) restriction fragment length polymorphism (RFLP) analysis of total genomic DNA with a repetitive DNA probe, and (v) detection and partial characterization of plasmid DNA. Fatty acid analysis identified all pathogenic strains as X. campestris, but did not consistently identify all the strains as X. campestris pv. vitians. The rep-PCR fingerprints and ITS1 sequences of all pathogenic X. campestris pv. vitians strains examined were identical, and distinct from those of the other X. campestris pathovars. Thus, these characteristics did not reveal genetic diversity among X. campestris pv. vitians strains, but did allow for differentiation of X. campestris pathovars. Genetic diversity among X. campestris pv. vitians strains was revealed by RFLP analysis with a repetitive DNA probe and by characterization of plasmid DNA. This diversity was greatest among strains from different geographical regions, although diversity among strains from the same location also was detected. The results of this study suggest that these X. campestris pv. vitians strains are not clonal, but comprise a relatively homogeneous group.


Biology ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 62 ◽  
Author(s):  
Lombardo ◽  
Fila ◽  
Lombardo ◽  
Epifani ◽  
III ◽  
...  

The primary impetus behind this research was to provide a boost to the characterization of the Italian olive biodiversity by acquiring reliable and homogeneous data over the course of an eight-year trial on the floral and fruiting biology of 120 molecularly analyzed cultivars, most of which have either low or very low diffusion. The obtained data highlighted a considerable variability to almost all of the analyzed parameters, which given the uniformity of environment and crop management was indicative of a large genetic variability in the accessions under observation, as confirmed through the molecular analysis. Several cases of synonymy were reported for the first time, even among plants cultivated in different regions, whilst all of the varieties examined, with only one exception, showed very low percentages of self-fruit-set, indicating a need for the employment of suitable pollinator plants. Eventually, a fitted model allowed us to evaluate the clear effects of the thermal values on blossoming, particularly in the months of March and April, whereas the climatic conditions during the flowering time had only a modest effect on its duration.


2019 ◽  
Vol 9 (7) ◽  
pp. 904-913
Author(s):  
Bing Yan ◽  
Ruining Liang ◽  
Meng Ji ◽  
Qi-Qige Wuyun ◽  
Weijun Guan ◽  
...  

Of all the significant researches that have taken place in isolation, culture and characterization of mesenchymal stem cells (MSCs), the field of kidney-derived mesenchymal stem cells (KMSCs) in Tibetan mastiff is still a blank. Therefore, the purpose of this study is to isolate, culture and characterize the Tibetan mastiff KMSCs. The KMSCs were successfully isolated from one-day year old Tibetan mastiff kidney, cultured for 16 passages and distinguished by two methods: immunofluorescence staining and RT-PCR. The Tibetan mastiff KMSCs expressed specific surface marker genes (VIM, CD44, FN1, CD90, CD109, CD73, FN1) and kidney marker gene PAX2. The proliferation ability of Tibetan mastiff KMSCs was measured through cell count and clonality. Furthermore, cells differentiated into different cell types (hepatocellular cells, osteogenic cells, adipogenic cells and chondrogenic cells) under special induced medium, and the marker genes of induced cells were identified with Immunofluorescence staining and RT-PCR. All of these results indicated that the Tibetan mastiff KMSCs were obtained successfully, which possessed certain characteristics of multipotent stem cells. Therefore, MSCs in Tibetan mastiff kidney hold potential for clinical applications for regenerative therapy and their further studies are waiting to be required to investigate their functions.


Author(s):  
Rajwent Singh ◽  
A.K. Arora ◽  
T.S. Rai ◽  
Mudit Chandra

Background: Group B streptococcus (GBS) or Streptococcus agalactiae is an important pathogen associated with bovine mastitis. The organism is also of public health consequences and may cause variety of infections ranging from neonatal sepsis, pneumonia and meningitis to localized infections and urinary tract infection or arthritisin adult humans. Widespread use of antibiotics in veterinary medicine has led to development of resistance among the pathogens. So there is need for surveillance of antimicrobial resistance to ensure effective treatment. Methods: Milk samples collected from mastitis affected animals were processed for isolation of Streptococcus agalactiae. The isolates were tested for antimicrobial susceptibility. Molecular characterisation was carried out by PCR to study the occurrence of resistance marker genes and virulence marker genes. RAPD was carried out to study genetic diversity among the isolates. Result: Six isolates of S. agalactiae were obtained from 182 milk samples. Highest resistance was observed against co-trimoxazole and tetracycline followed by ampicillin. tetM gene and tetO genes could be amplified in four and three isolates, respectively. None of the isolates showed amplification for ermA, ermB, mefA and mefE genes. Three isolates were positive for the five virulence genes tested (glnA, cfb, hylB, scaA and cyl). RAPD analysis demonstrated great intraspecific genetic diversity among the streptococcal isolates.


Genome ◽  
2006 ◽  
Vol 49 (4) ◽  
pp. 289-296 ◽  
Author(s):  
Ervin D Nagy ◽  
István Molnár ◽  
Annamária Schneider ◽  
Géza Kovács ◽  
Márta Molnár-Láng

The short interspersed nuclear element (SINE), Au, was used to develop sequence-specific amplified polymorphism (S-SAP) markers for U- and M-genome chromosomes. The markers were localized using Triticum aestivum (wheat) – Aegilops geniculata and wheat – Aegilops biuncialis disomic chromosome addition lines. Thirty-seven markers distributed over 6 U and 6 M chromosomes were produced. A genetic diversity study carried out on 37 accessions from Ae. biuncialis, Ae. comosa, Ae. geniculata, and Ae. umbellulata suggested that Ae. biuncialis have arisen from its diploid ancestors more recently than Ae. geniculata. Several earlier studies indicated that the M genomes in polyploid Aegilops species had accumulated substantial rearrangements, whereas the U genomes remained essentially unmodified. However, this cannot be attributed to the preferential insertion of retroelements into the M genome chromosomes. Fourteen markers from a total of 8 chromosomes were sequenced; 3 markers were similar to known plant genes, 1 was derived from a long terminal repeat (LTR) retrotransposon, and 10 markers did not match to any known DNA sequences, suggesting that they were located in the highly variable intergenic regions.Key words: Aegilops, U and M genomes, S-SAP, genetic diversity, sequence characterization.


2015 ◽  
Vol 46 (1) ◽  
pp. 119-125
Author(s):  
Cristiane Silva Chitarra ◽  
Mayara Inácio Vincenzi da Silva ◽  
Laila Natasha Santos Brandão ◽  
Francielle Cristina Kagueyama ◽  
Stefhano Luis Candido ◽  
...  

ABSTRACT: Swine respiratory diseases such as atrophic rhinitis and bronchopneumonia caused by Pasteurella (P.) multocida cause important economic losses to the modern swine industry. The purpose of this study was to characterize P. multocida strains isolated from swine lungs by RAPD (Randomly Amplified Polymorphic DNA) to demonstrate their genetic diversity. Ninety-four samples of fragments from lungs with pneumonia and sixty one samples without pneumonia were collected in slaughterhouses in Mato Grosso during the period from December 2009 to March 2010. Clinical cases in 2012 and 2013 were also included in this study. Among the lung fragments with macroscopic lesions, without macroscopic lesions and clinical samples, 40.42%, 4.49% and 100% were positive for P. multocida, respectively. Bacterial identification culturing was confirmed by PCR (polymerase chain reaction) by means of the amplification of the gene kmt1. RAPD technique was performed for 46 isolates, and in every isolate, a total of 7 to 11 amplification bands were detected, composed of 8 clusters based on genetic similarity. Thus, treatment, control and preventive measures should consider the genetic diversity of P. multocida populations in swine herds in order to improve the development of new protocols to produce antimicrobials and vaccines.


2020 ◽  
Author(s):  
Quentin Carradec ◽  
Julie Poulain ◽  
Emilie Boissin ◽  
Benjamin CC Hume ◽  
Christian R Voolstra ◽  
...  

AbstractMolecular characterization of the coral host and the microbial assemblages associated with it (referred to as the coral holobiont) is currently undertaken via marker gene sequencing. This requires bulky instruments and controlled laboratory conditions which are impractical for environmental experiments in remote areas. Recent advances in sequencing technologies now permit rapid sequencing in the field; however, development of specific protocols and pipelines for the effective processing of complex microbial systems are currently lacking. Here, we used a combination of 3 marker genes targeting the coral animal host, its symbiotic alga, and the associated bacterial microbiome to characterize 60 coral colonies collected and processed in situ, during the Tara Pacific expedition. We used Oxford Nanopore Technologies to sequence marker gene amplicons and developed bioinformatics pipelines to analyze nanopore reads on a laptop, obtaining results in less than 24 hours. Reef scale network analysis of coral-associated bacteria reveals broadly distributed taxa, as well as host-specific associations. Protocols and tools used in this work may be applicable for rapid coral holobiont surveys, immediate adaptation of sampling strategy in the field, and to make informed and timely decisions in the context of the current challenges affecting coral reefs worldwide.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Quentin Carradec ◽  
Julie Poulain ◽  
Emilie Boissin ◽  
Benjamin C. C. Hume ◽  
Christian R. Voolstra ◽  
...  

Abstract Molecular characterization of the coral host and the microbial assemblages associated with it (referred to as the coral holobiont) is currently undertaken via marker gene sequencing. This requires bulky instruments and controlled laboratory conditions which are impractical for environmental experiments in remote areas. Recent advances in sequencing technologies now permit rapid sequencing in the field; however, development of specific protocols and pipelines for the effective processing of complex microbial systems are currently lacking. Here, we used a combination of 3 marker genes targeting the coral animal host, its symbiotic alga, and the associated bacterial microbiome to characterize 60 coral colonies collected and processed in situ, during the Tara Pacific expedition. We used Oxford Nanopore Technologies to sequence marker gene amplicons and developed bioinformatics pipelines to analyze nanopore reads on a laptop, obtaining results in less than 24 h. Reef scale network analysis of coral-associated bacteria reveals broadly distributed taxa, as well as host-specific associations. Protocols and tools used in this work may be applicable for rapid coral holobiont surveys, immediate adaptation of sampling strategy in the field, and to make informed and timely decisions in the context of the current challenges affecting coral reefs worldwide.


2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
M. K. Rajesh ◽  
K. Samsudeen ◽  
P. Rejusha ◽  
C. Manjula ◽  
Shafeeq Rahman ◽  
...  

The coconut palm is versatile in its adaptability to a wide range of soil and climatic conditions. A long history of its cultivation has resulted in development of many ecotypes, which are adapted to various agro-eco factors prevalent in a particular region. These ecotypes usually are known by the location where they are grown. It is important to explore such adaptation in the coconut population for better utilization of these ecotypes in coconut breeding programs. The aim of the present study was to identify the genetic diversity of the Bedakam and Annur ecotypes of coconut and compare these ecotypes with predominant West Coast Tall (WCT) populations, from which they are presumed to have been derived, using microsatellite markers. All the 17 microsatellite markers used in the study revealed 100% polymorphism. The clustering analysis showed that Annur and Bedakam ecotypes were two separate and distinct populations compared to WCT. It was also evident from the clustering that Annur ecotype was closer to WCT than Bedakam ecotype.


2021 ◽  
Vol 28 (1) ◽  
pp. e17867
Author(s):  
Pedro Eduardo Romero ◽  
Camila Castillo-Vilcahuaman

Genetic diversity is an important component of biodiversity, and it is crucial for current efforts to protect and sustainably manage several organisms and habitats. As far as we know, there is only one work describing Peruvian genetic information stored in public databases. We aimed to update this previous work searching in four public databases that stored digital sequence information: Nucleotide, BioProject, PATRIC, BOLD. With this information, we comment on the contribution of Peruvian institutions during recent years. In Nucleotide, the largest database, Bacteria are the most sequenced organisms by Peruvian institutions (70.60%), pathogenic bacteria such as Pasteurella multocida, Neisseria meningitidis, and Vibrio parahaemolyticus were the most abundant. We found no sequence records from the Archaea domain. In BioProject, the most common sequence belongs to Salmonella enterica subsp. enterica serovar Infantis. In PATRIC, a database of pathogenic agents, Mycobacterium tuberculosis and Yersinia pestis had the highest number of entries. Finally, in BOLD, an exclusively Eukaryotic database, Chordata (Aves and Actinopterygii), Angiospermae, and Arthropoda (Insecta, and Arachnida) were the most frequent records. Our results would indicate research preferences of Peruvian institutions, focusing on infectious diseases and some Eukaryotic phyla. Although there has been a significant increase of DNA information submitted by Peruvian institutions since the last report, the genetic diversity reflected in these databases remains inconsistent with the diversity in the country. More efforts must be made to obtain genetic information from more underestimated taxonomic groups and to promote more genetic research in regional Peruvian institutions.


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