Asymmetrical coexistence of Nosema ceranae and Nosema apis in honeybees. J Invertebr Pathol

Department of Biology, College of Life Science, University of Maryland, College Park, MD, USA
Journal of Invertebrate Pathology (Impact Factor: 2.11). 06/2009; 101(3):204-209. DOI: 10.1016/j.jip.2009.05.012
Source: PubMed


Globalization has provided opportunities for parasites/pathogens to cross geographic boundaries and expand to new hosts. Recent studies showed that Nosema ceranae, originally considered a microsporidian parasite of Eastern honey bees, Apis cerana, is a disease agent of nosemosis in European honey bees, Apis mellifera, along with the resident species, Nosema apis. Further studies indicated that disease caused by N. ceranae in European honey bees is far more prevalent than that caused by N. apis. In order to gain more insight into the epidemiology of Nosema parasitism in honey bees, we conducted studies to investigate infection of Nosema in its original host, Eastern honey bees, using conventional PCR and duplex real time quantitative PCR methods. Our results showed that A. cerana was infected not only with N. ceranae as previously reported [Fries, I., Feng, F., Silva, A.D., Slemenda, S.B., Pieniazek, N.J., 1996. Nosema ceranae n. sp. (Microspora, Nosematidae), morphological and molecular characterization of a microsporidian parasite of the Asian honey bee Apis cerana (Hymenoptera, Apidae). Eur. J. Protistol. 32, 356–365], but also with N. apis. Both microsporidia produced single and mixed infections. Overall and at each location alone, the prevalence of N. ceranae was higher than that of N. apis. In all cases of mixed infections, the number of N. ceranae gene copies (corresponding to the parasite load) significantly out numbered those of N. apis. Phylogenetic analysis based on a variable region of small subunit ribosomal RNA (SSUrRNA) showed four distinct clades of N. apis and five clades of N. ceranae and that geographical distance does not appear to influence the genetic diversity of Nosema populations. The results from this study demonstrated that duplex real-time qPCR assay developed in this study is a valuable tool for quantitative measurement of Nosema and can be used to monitor the progression of microsprodian infections of honey bees in a timely and cost efficient manner.

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    • "Single-and co-infections of N. ceranae and N. apis occur in both A. mellifera and A. cerana colonies with higher prevalence of N. ceranae (Chen et al., 2009; Klee et al., 2007). This may be associated with faster reproduction of N. ceranae, geographic isolation or the inability of host immune system to cope with N. ceranae infection (Chen et al., 2009; Martin- Hernandez et al., 2011; Shutler et al., 2014). Nosema alters aspects of both honey bee behavior and physiology (Hassanein, 1951; Goblirsch et al., 2013; Higes et al., 2010). "
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    ABSTRACT: The objectives of this study were to quantify the costs and benefits of co-parasitism with Varroa (Varroa destructor Anderson and Trueman) and Nosema (Nosema ceranae Fries and Nosema apis Zander) on honey bees (Apis mellifera L.) with different defense levels. Newly-emerged worker bees from either high-mite-mortality-rate (high-MMR) bees or low-mite-mortality-rate (low-MMR) bees were confined in forty bioassay cages which were either inoculated with Nosema spores [Nosema (+) group] or were left un-inoculated [Nosema (-) group]. Caged-bees were then inoculated with Varroa mites [Varroa (+) group] or were left untreated [Varroa (+) group]. This established four treatment combinations within each Nosema treatment group: (1) low-MMR Varroa (-), (2) high-MMR Varroa (-), (3) low-MMR Varroa (+) and (4) high-MMR Varroa (+), each with five replicates. Overall mite mortality in high-MMR bees (0.12±0.02 mites per day) was significantly greater than in the low-MMR bees (0.06±0.02 mites per day). In the Nosema (-) groups bee mortality was greater in high-MMR bees than low-MMR bees but only when bees had a higher mite burden. Overall, high-MMR bees in the Nosema (-) group showed greater reductions in mean abundance of mites over time compared with low-MMR bees, when inoculated with additional mites. However, high-MMR bees could not reduce mite load as well as in the Nosema (-) group when fed with Nosema spores. Mean abundance of Nosema spores in live bees and dead bees of both strains of bees was significantly greater in the Nosema (+) group. Molecular analyses confirmed the presence of both Nosema species in inoculated bees but N. ceranae was more abundant than N. apis and unlike N. apis increased over the course of the experiment. Collectively, this study showed differential mite mortality rates among different genotypes of bees, however, Nosema infection restrained Varroa removal success in high-MMR bees. Copyright © 2015. Published by Elsevier Inc.
    Journal of Invertebrate Pathology 08/2015; 132. DOI:10.1016/j.jip.2015.07.019 · 2.11 Impact Factor
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    • "In Turkey , the presence of N . apis was confirmed earlier ( Aydin et al . , 2005 ; Muz et al . , 2010 ; Whitaker et al . , 2010 ) . Using qPCR , we detected N . apis in only three colonies , indicating the replacement of N . apis by N . ceranae in many regions included in this study . Worldwide , N . ceranae is now far more common than N . apis ( Chen et al . , 2009 ; Valera et al . , 2011 ; Yoshiyama and Kimura , 2011 ) ."
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    ABSTRACT: Honey bees face numerous biotic threats from viruses to bacteria, fungi, protists, and mites. Here we describe a thorough analysis of microbes harbored by worker honey bees collected from field colonies in geographically distinct regions of Turkey. Turkey is one of the World's most important centers of apiculture, harboring five subspecies of Apis mellifera L., approximately 20% of the honey bee subspecies in the world. We use deep ILLUMINA-based RNA sequencing to capture RNA species for the honey bee and a sampling of all non-endogenous species carried by bees. After trimming and mapping these reads to the honey bee genome, approximately 10% of the sequences (9-10 million reads per library) remained. These were then mapped to a curated set of public sequences containing ca. Sixty megabase-pairs of sequence representing known microbial species associated with honey bees. Levels of key honey bee pathogens were confirmed using quantitative PCR screens. We contrast microbial matches across different sites in Turkey, showing new country recordings of Lake Sinai virus, two Spiroplasma bacterium species, symbionts Candidatus Schmidhempelia bombi, Frischella perrara, Snodgrassella alvi, Gilliamella apicola, Lactobacillus spp.), neogregarines, and a trypanosome species. By using metagenomic analysis, this study also reveals deep molecular evidence for the presence of bacterial pathogens (Melissococcus plutonius, Paenibacillus larvae), Varroa destructor-1 virus, Sacbrood virus, and fungi. Despite this effort we did not detect KBV, SBPV, Tobacco ringspot virus, VdMLV (Varroa Macula like virus), Acarapis spp., Tropilaeleps spp. and Apocephalus (phorid fly). We discuss possible impacts of management practices and honey bee subspecies on microbial retinues. The described workflow and curated microbial database will be generally useful for microbial surveys of healthy and declining honey bees.
    Frontiers in Genetics 03/2015; 6:100. DOI:10.3389/fgene.2015.00100
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    • "Freshly isolated spores were used immediately for each experimental treatment. Nosema isolates were confirmed to be pure species using PCR diagnoses (Huang and Solter, 2013; Chen et al., 2009). "
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    ABSTRACT: Nosema ceranae infection is ubiquitous in western honey bees, Apis mellifera, in the United States and the pathogen has apparently replaced Nosema apis in colonies nationwide. Displacement of N. apis suggests that N. ceranae has competitive advantages but N. ceranae was significantly less infective and less virulent than N. apis in commercially available lineages of honey bees in studies conducted in Illinois and Texas. At 5 days post eclosion, the most susceptible age of adult bees tested, the mean ID50 for N. apis was 359 spores compared to 3217 N. ceranae spores, a nearly 9-fold difference. Infectivity of N. ceranae was also lower than N. apis for 24-h and 14-day worker bees. N. ceranae was less infective than reported in studies using European strains of honey bees, while N. apis infectivity, tested in the same cohort of honey bees, corresponded to results reported globally from 1972 to 2010. Mortality of worker bees was similar for both pathogens at a dosage of 50 spores and was not different from the uninfected controls, but was significantly higher for N. apis than N. ceranae at dosages ⩾500 spores. Our results provide comparisons for evaluating research using different ages of bees and pathogen dosages and clarify some controversies. In addition, comparisons among studies suggest that the mixed lineages of US honey bees may be less susceptible to N. ceranae infections than are European bees or that the US isolates of the pathogen are less infective and less virulent than European isolates.
    Journal of Invertebrate Pathology 10/2014; 124. DOI:10.1016/j.jip.2014.10.006 · 2.11 Impact Factor
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