scholarly journals Characterization of CRISPR Spacer and Protospacer Sequences in Paenibacillus larvae and Its Bacteriophages

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 459
Author(s):  
Casey Stamereilers ◽  
Simon Wong ◽  
Philippos K. Tsourkas
Keyword(s):  
Comparative Studies ◽  
Point Mutations ◽  
Paenibacillus Larvae ◽  
Bacterial Disease ◽  
American Foulbrood ◽  
Antibiotic Resistant ◽  
Future Studies ◽  
Complete Genomes ◽  
Point Of Reference

The bacterium Paenibacillus larvae is the causative agent of American foulbrood, the most devastating bacterial disease of honeybees. Because P. larvae is antibiotic resistant, phages that infect it are currently used as alternative treatments. However, the acquisition by P. larvae of CRISPR spacer sequences from the phages could be an obstacle to treatment efforts. We searched nine complete genomes of P. larvae strains and identified 714 CRISPR spacer sequences, of which 384 are unique. Of the four epidemiologically important P. larvae strains, three of these have fewer than 20 spacers, while one strain has over 150 spacers. Of the 384 unique spacers, 18 are found as protospacers in the genomes of 49 currently sequenced P. larvae phages. One P. larvae strain does not have any protospacers found in phages, while another has eight. Protospacer distribution in the phages is uneven, with two phages having up to four protospacers, while a third of phages have none. Some phages lack protospacers found in closely related phages due to point mutations, indicating a possible escape mechanism. This study serve a point of reference for future studies on the CRISPR-Cas system in P. larvae as well as for comparative studies of other phage–host systems.

scholarly journals Isolation and Characterization of Phages Active against Paenibacillus larvae Causing American Foulbrood in Honeybees in Poland

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1217
Author(s):  
Ewa Jończyk-Matysiak ◽  
Barbara Owczarek ◽  
Ewa Popiela ◽  
Kinga Świtała-Jeleń ◽  
Paweł Migdał ◽  
...  
Keyword(s):  
Biological Properties ◽  
Field Application ◽  
Paenibacillus Larvae ◽  
American Foulbrood ◽  
Isolation And Characterization ◽  
Safety Studies ◽  
Spreading Disease ◽  
Intended Use ◽  
Antibacterial Potential

The aim of this study was the isolation and characterization, including the phage effect on honeybees in laboratory conditions, of phages active against Paenibacillus larvae, the causative agent of American Foulbrood—a highly infective and easily spreading disease occurring in honeybee larva, and subsequently the development of a preparation to prevent and treat this dangerous disease. From the tested material (over 2500 samples) 35 Paenibacillus spp. strains were obtained and used to search for phages. Five phages specific to Paenibacillus were isolated and characterized (ultrastructure, morphology, biological properties, storage stability, and genome sequence). The characteristics were performed to obtain knowledge of their lytic potential and compose the final phage cocktail with high antibacterial potential and intended use of future field application. Preliminary safety studies have also been carried out on healthy bees, which suggest that the phage preparation administered is harmless.

Biochemical characterization of different genotypes of Paenibacillus larvae subsp. larvae, a honey bee bacterial pathogen

Microbiology ◽  
2004 ◽  
Vol 150 (7) ◽  
pp. 2381-2390 ◽  
Author(s):  
Sandra Neuendorf ◽  
Kati Hedtke ◽  
Gerhard Tangen ◽  
Elke Genersch
Keyword(s):  
Honey Bee ◽  
Reference Strain ◽  
Biochemical Characterization ◽  
Bacterial Pathogen ◽  
Paenibacillus Larvae ◽  
Bacterial Disease ◽  
System A ◽  
Biolog System ◽  
Far Future

Paenibacillus larvae subsp. larvae (P. l. larvae) is the aetiological agent of American foulbrood (AFB), the most virulent bacterial disease of honey bee brood worldwide. In many countries AFB is a notifiable disease since it is highly contagious, in most cases incurable and able to kill affected colonies. Genotyping of field isolates of P. l. larvae revealed at least four genotypes (AB, Ab, ab and α B) present in Germany which are genotypically different from the reference strain DSM 7030. Therefore, based on these data, five different genotypes of P. l. larvae are now identified with genotype AB standing out with a characteristic brown-orange and circled two-coloured colony morphology. Analysing the metabolic profiles of three German genotypes (AB, Ab and ab) as well as of the reference strain using the Biolog system, a characteristic biochemical fingerprint could be obtained for each strain. Cluster analysis showed that while genotypes Ab, ab and the reference strain DSM 7030 are rather similar, genotype AB is clearly different from the others. Analysis of all isolates for plasmid DNA revealed two different plasmids present only in isolates belonging to genotype AB. Therefore, genotype AB is remarkable in all aspects analysed so far. Future analysis will show whether or not these differences will expand to differences in virulence.

scholarly journals Antimicrobial Resistance Genes in Bacteria Isolated From Japanese Honey, and Their Potential for Conferring Macrolide and Lincosamide Resistance in the American Foulbrood Pathogen Paenibacillus larvae

2021 ◽  
Vol 12 ◽  
Author(s):  
Mariko Okamoto ◽  
Masahiko Kumagai ◽  
Hiroyuki Kanamori ◽  
Daisuke Takamatsu
Keyword(s):  
Resistance Genes ◽  
Macrolide Resistance ◽  
Paenibacillus Larvae ◽  
Resistant Bacteria ◽  
Bacterial Disease ◽  
American Foulbrood ◽  
Mobilizable Plasmid ◽  
Antimicrobial Resistance Genes ◽  
Cloned Gene ◽  
Lincomycin Resistance

American foulbrood (AFB) is the most serious bacterial disease of honey bee brood. Spores of the causative agent Paenibacillus larvae are ingested by bee larvae via brood foods and germinated cells proliferate in the larval midgut. In Japan, a macrolide antibiotic, tylosin, is used as the approved prophylactic for AFB. Although tylosin-resistant P. larvae has yet to be found in Japan, it may emerge in the future through the acquisition of macrolide resistance genes from other bacteria, and bacteria latent in brood foods, such as honey, may serve as a source of resistance genes. In this study, to investigate macrolide resistance genes in honey, we attempted to isolate tylosin-resistant bacteria from 53 Japanese honey samples and obtained 209 isolates from 48 samples in the presence of 1 μg/ml of tylosin. All isolates were Gram-positive spore-forming bacteria mainly belonging to genera Bacillus and Paenibacillus, and 94.3% exhibited lower susceptibility to tylosin than Japanese P. larvae isolates. Genome analysis of 50 representative isolates revealed the presence of putative macrolide resistance genes in the isolates, and some of them were located on mobile genetic elements (MGEs). Among the genes on MGEs, ermC on the putative mobilizable plasmid pJ18TS1mac of Oceanobacillus strain J18TS1 conferred tylosin and lincomycin resistance to P. larvae after introducing the cloned gene using the expression vector. Moreover, pJ18TS1mac was retained in the P. larvae population for a long period even under non-selective conditions. This suggests that bacteria in honey is a source of genes for conferring tylosin resistance to P. larvae; therefore, monitoring of bacteria in honey may be helpful to predict the emergence of tylosin-resistant P. larvae and prevent the selection of resistant strains.

scholarly journals American foulbrood and its causative agent, Paenibacillus larvae, in New Zealand’s registered hives and apiaries

2021 ◽  
Author(s):  
◽  
Samantha Amy Montrose Graham
Keyword(s):  
New Zealand ◽  
Statistical Analysis ◽  
Genetic Analysis ◽  
Honey Bee ◽  
Causative Agent ◽  
Management Strategies ◽  
Paenibacillus Larvae ◽  
Bacterial Disease ◽  
American Foulbrood ◽  
Infection Rates

<p>Though the honey bee (Apis mellifera) is exposed to an extensive diversity of parasites and pathogens from multiple kingdoms, few are as devastating as American foulbrood. American foulbrood is a highly contagious bacterial disease, of which the causative agent (bacterium Paenibacillus larvae) infects honey bee brood through the ingestion of its spores, ultimately leading to the death of the infected larva and the collapse of the infected hive. Paenibacillus larvae’s genotypes (ERIC I-IV) exhibit differing ‘killing time’ of infected larvae, resulting in different larval and colony level virulence of the disease within hives.  American foulbrood is found in New Zealand’s registered hives, and poses a threat to the country’s apiculture industry. The first objective of this thesis was to perform a genetic analysis on New Zealand’s P. larvae field strains using the well-established methodology of rep-PCR with MBO REP1 primers. A total of 172 bacteria isolates were gathered from registered hives from 2011 to 2014 and examined. The MBO REP1 primer identifies the ‘beta’ genetic subgroups of P. larvae. By identifying beta subgroups, the ERIC genotypes that are present in New Zealand can also be concluded. The genetic analysis of P. larvae using rep-PCR is a first for New Zealand, and appears to be a first for Australasia. The second objective of this thesis was to conduct a temporal and geographical statistical analysis on American foulbrood infection rate trends in New Zealand’s national and regional, divided into seven regions, registered hives and apiaries from 1994 to 2013.  The genetic analysis of P. larvae detected three ‘beta’ genotypic subgroups: B, b, and Б. From these findings it was concluded that ERIC I and ERIC II are present in New Zealand. Previous to my findings, subgroup B and Б and ERIC II genotype had not been recorded outside of Europe. The statistical analysis reported that American foulbrood infection rates were significantly decreasing nationally. Results also reported that four of the seven regions’ infection rates were significantly decreasing, whilst three regions were significantly increasing.  Conclusions on the subgroups and genotypes present in New Zealand gives the first insight to the virulence and occurrence of P. larvae strains. Additionally, the use of rep-PCR for the genetic analysis of P. larvae enables this thesis to contribute to the increasing knowledge on American foulbrood. By examining the temporal and geographic dynamics of American foulbrood, the results allow for the evaluation of current management strategies and the most recent understanding on the national and regional infection rates of the disease.</p>

scholarly journals Complete Genome Sequences of 18 Paenibacillus larvae Phages from the Western United States

2018 ◽  
Vol 7 (13) ◽  
Author(s):  
Bryan D. Merrill ◽  
Christopher P. Fajardo ◽  
Jared A. Hilton ◽  
Ashley M. Payne ◽  
Andy T. Ward ◽  
...  
Keyword(s):  
United States ◽  
Causative Agent ◽  
Complete Genome ◽  
Western United States ◽  
Paenibacillus Larvae ◽  
American Foulbrood ◽  
Genome Sequences ◽  
Content Type ◽  
Brigham Young University ◽  
Complete Genomes

We present here the complete genomes of 18 phages that infect Paenibacillus larvae, the causative agent of American foulbrood in honeybees. The phages were isolated between 2014 and 2016 as part of an undergraduate phage discovery course at Brigham Young University.

scholarly journals Complete Genome Sequences of Paenibacillus larvae Phages BN12, Dragolir, Kiel007, Leyra, Likha, Pagassa, PBL1c, and Tadhana

2018 ◽  
Vol 6 (24) ◽  
Author(s):  
Jamison K. Walker ◽  
Bryan D. Merrill ◽  
Jordan A. Berg ◽  
Aziza Dhalai ◽  
Douglas W. Dingman ◽  
...  
Keyword(s):  
Causative Agent ◽  
Complete Genome ◽  
Paenibacillus Larvae ◽  
American Foulbrood ◽  
Genome Sequences ◽  
Content Type ◽  
Complete Genomes ◽  
The Usa

ABSTRACT We present here the complete genomes of eight phages that infect Paenibacillus larvae, the causative agent of American foulbrood in honeybees. Phage PBL1c was originally isolated in 1984 from a P. larvae lysogen, while the remaining phages were isolated in 2014 from bee debris, honeycomb, and lysogens from three states in the USA.

scholarly journals American foulbrood and its causative agent, Paenibacillus larvae, in New Zealand’s registered hives and apiaries

2021 ◽  
Author(s):  
◽  
Samantha Amy Montrose Graham
Keyword(s):  
New Zealand ◽  
Statistical Analysis ◽  
Genetic Analysis ◽  
Honey Bee ◽  
Causative Agent ◽  
Management Strategies ◽  
Paenibacillus Larvae ◽  
Bacterial Disease ◽  
American Foulbrood ◽  
Infection Rates

<p>Though the honey bee (Apis mellifera) is exposed to an extensive diversity of parasites and pathogens from multiple kingdoms, few are as devastating as American foulbrood. American foulbrood is a highly contagious bacterial disease, of which the causative agent (bacterium Paenibacillus larvae) infects honey bee brood through the ingestion of its spores, ultimately leading to the death of the infected larva and the collapse of the infected hive. Paenibacillus larvae’s genotypes (ERIC I-IV) exhibit differing ‘killing time’ of infected larvae, resulting in different larval and colony level virulence of the disease within hives.  American foulbrood is found in New Zealand’s registered hives, and poses a threat to the country’s apiculture industry. The first objective of this thesis was to perform a genetic analysis on New Zealand’s P. larvae field strains using the well-established methodology of rep-PCR with MBO REP1 primers. A total of 172 bacteria isolates were gathered from registered hives from 2011 to 2014 and examined. The MBO REP1 primer identifies the ‘beta’ genetic subgroups of P. larvae. By identifying beta subgroups, the ERIC genotypes that are present in New Zealand can also be concluded. The genetic analysis of P. larvae using rep-PCR is a first for New Zealand, and appears to be a first for Australasia. The second objective of this thesis was to conduct a temporal and geographical statistical analysis on American foulbrood infection rate trends in New Zealand’s national and regional, divided into seven regions, registered hives and apiaries from 1994 to 2013.  The genetic analysis of P. larvae detected three ‘beta’ genotypic subgroups: B, b, and Б. From these findings it was concluded that ERIC I and ERIC II are present in New Zealand. Previous to my findings, subgroup B and Б and ERIC II genotype had not been recorded outside of Europe. The statistical analysis reported that American foulbrood infection rates were significantly decreasing nationally. Results also reported that four of the seven regions’ infection rates were significantly decreasing, whilst three regions were significantly increasing.  Conclusions on the subgroups and genotypes present in New Zealand gives the first insight to the virulence and occurrence of P. larvae strains. Additionally, the use of rep-PCR for the genetic analysis of P. larvae enables this thesis to contribute to the increasing knowledge on American foulbrood. By examining the temporal and geographic dynamics of American foulbrood, the results allow for the evaluation of current management strategies and the most recent understanding on the national and regional infection rates of the disease.</p>

scholarly journals Genomic Analysis of 48 Paenibacillus larvae Bacteriophages

Viruses ◽  
2018 ◽  
Vol 10 (7) ◽  
pp. 377 ◽  
Author(s):  
Casey Stamereilers ◽  
Christopher Fajardo ◽  
Jamison Walker ◽  
Katterinne Mendez ◽  
Eduardo Castro-Nallar ◽  
...  
Keyword(s):  
Genomic Analysis ◽  
Dna Packaging ◽  
Paenibacillus Larvae ◽  
Bacterial Disease ◽  
Replication Proteins ◽  
Antibiotic Resistant ◽  
Distinct Cluster ◽  
Antibiotic Resistant Bacterium ◽  
Packaging Strategy ◽  
Assembly Proteins

The antibiotic-resistant bacterium Paenibacillus larvae is the causative agent of American foulbrood (AFB), currently the most destructive bacterial disease in honeybees. Phages that infect P. larvae were isolated as early as the 1950s, but it is only in recent years that P. larvae phage genomes have been sequenced and annotated. In this study we analyze the genomes of all 48 currently sequenced P. larvae phage genomes and classify them into four clusters and a singleton. The majority of P. larvae phage genomes are in the 38–45 kbp range and use the cohesive ends (cos) DNA-packaging strategy, while a minority have genomes in the 50–55 kbp range that use the direct terminal repeat (DTR) DNA-packaging strategy. The DTR phages form a distinct cluster, while the cos phages form three clusters and a singleton. Putative functions were identified for about half of all phage proteins. Structural and assembly proteins are located at the front of the genome and tend to be conserved within clusters, whereas regulatory and replication proteins are located in the middle and rear of the genome and are not conserved, even within clusters. All P. larvae phage genomes contain a conserved N-acetylmuramoyl-l-alanine amidase that serves as an endolysin.

Identification and Treatment of European Foulbrood in Honey Bee Colonies

EDIS ◽  
2020 ◽  
Vol 2020 (1) ◽  
pp. 7
Author(s):  
Catherine M. Mueller ◽  
Cameron Jack ◽  
Ashley N. Mortensen ◽  
Jamie D. Ellis
Keyword(s):  
Honey Bee ◽  
Bacterial Disease ◽  
American Foulbrood ◽  
Fact Sheet ◽  
European Foulbrood ◽  
Bee Colonies ◽  
Honey Bee Larvae

European foulbrood is a bacterial disease that affects Western honey bee larvae. It is a concern to beekeepers everywhere, though it is less serious than American foulbrood because it does not form spores, which means that it can be treated. This 7-page fact sheet written by Catherine M. Mueller, Cameron J. Jack, Ashley N. Mortensen, and Jamie Ellis and published by the UF/IFAS Entomology and Nematology Department describes the disease and explains how to identify it to help beekeepers manage their colonies effectively and prevent the spread of both American and European foulbrood.https://edis.ifas.ufl.edu/in1272

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