Search Results (48)

Despite the advantages associated with social living, one of the main costs of sociality is an increased exposure to pathogens. For honey bees, these costs are artificially inflated in the unnatural and densely crowded apiary environments common to commercial operations. Modern beekeeping practices have not only exacerbated these problems but have also led to the introduction of novel parasites. One newly introduced parasite of honey bees is Nosema ceranae, a unicellular, spore-forming gut parasite that has achieved a cosmopolitan distribution in recent decades. N. ceranae is widely considered a dangerous pathogen for both individual bees and colonies as a whole. The parasite can infect every member of the honey bee colony; however, it remains to be understood whether there are asymmetrical contributions to spreading N. ceranae spores by particular castes. Here, we test the long-held suspicion that honey bee drones may be important for driving pathogen transmission between colonies in modern apiary settings. Although we find sparse evidence that N. ceranae infection causes enhanced transmission by changing drone or worker behavior, we do confirm that infected drones drift at high rates and are accepted by foreign colonies. Our results thus present a grim picture for enhanced diseases transmission by males in crowded apiaries. Full article

Nosemosis is a disease caused by microsporidia, which are strictly intracellular pathogens, currently considered to be most closely related to fungi. These microscopic parasites infect a variety of hosts, significantly affecting honeybees (Apis mellifera). Nosemosis is one of the most serious diseases of bees and is caused primarily by two species: Nosema apis and Nosema ceranae. This infection adversely affects the digestive tract of the bees, causes a reduction in their vitality, and can lead to the death of entire colonies. The diagnosis of nosemosis has undergone extensive development. Traditionally, the identification of microsporidia was performed by examination of bee digestive tract (macerated) by light microscopy. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) are expensive methods that require skilled personnel and were used only when high resolution was necessary. Modern methods, such as polymerase chain reaction (PCR), allow detection of infection at species and genotype levels, thereby increasing the accuracy of diagnosis. Despite advances in molecular techniques, research into nosemosis still faces challenges. This review focuses on a comparison of different diagnostic techniques and their pitfalls that can be integrated into strategies to combat nosemosis and protect the health of honeybee colonies. Full article

Honey bees are essential pollinators for the ecosystem and food crops. However, their health and survival face threats from both biotic and abiotic stresses. Fungi, microsporidia, and bacteria might significantly contribute to colony losses. Therefore, rapid and sensitive diagnostic tools are crucial for effective disease management. In this study, molecular assays were developed to quickly and efficiently detect the main honey bee pathogens: Nosema ceranae, Aspergillus flavus, Paenibacillus larvae, and Black queen cell virus. In this context, new primer pairs were designed for use in quantitative Real-time PCR (qPCR) reactions. Various protocols for extracting total nucleic acids from bee tissues were tested, indicating a CTAB-based protocol as the most efficient and cost-effective. Furthermore, excluding the head of the bee from the extraction, better results were obtained in terms of quantity and purity of extracted nucleic acids. These assays showed high specificity and sensitivity, detecting up to 250 fg of N. ceranae, 25 fg of P. larvae, and 2.5 pg of A. flavus DNA, and 5 pg of BQCV cDNA, without interference from bee DNA. These qPCR assays allowed pathogen detection within 3 h and at early stages of infection, supporting timely and efficient management interventions. Full article

One of the common causes of mass death in bee colonies is the infectious disease nosemosis, which is caused by two types of microsporidia, Nosema apis and Nosema ceranae. Of the many factors contributing to the spread of nosemosis, in this paper we consider the hybridization of subspecies of Apis mellifera L. In most of Russia, the native subspecies is the dark forest bee Apis mellifera mellifera, which is representative of the evolutionary lineage M. The export of bee packages and queens from the southern regions of Russia and other countries has led to the fragmentation of the range of these subspecies. First, we determined the maternal and paternal ancestry of 349 honey bee colonies across 12 beekeeping regions of Russia using the mitochondrial tRNAleu-COII locus and nine nuclear SSR markers (Ap243, 4a110, A024, A008, A43, A113, A088, Ap049, and A028). Among them, 140 colonies belonged to subspecies A. m. mellifera, 58 colonies were of hybrid origin, and 151 colonies belonged to evolutionary lineage C. Then, using microscopy and PCR analysis, we performed diagnostics of nosemosis in the studied colonies: N. apis was detected in 87 colonies, N. ceranae in 102 colonies, and coinfection was observed in 36 colonies. The results of our study indicate that the main reservoir of Nosema microsporidia was bees of evolutionary lineage C. Full article

Honeybee diseases are one of the most significant and most common causes of honeybee colonies’ weakness and death. An early diagnosis of subclinical infections is necessary to implement precautionary and control measures. Sampling debris from hive bottom boards is simple, non-invasive, and cheap. In this study, we collected winter debris samples in apiaries located in the continental part of Croatia. We used molecular methods, PCR and qPCR, for the first time to analyze those samples. Laboratory results were compared with the health condition and strength of honeybee colonies at an apiary in spring. Our study successfully identified the presence and quantity of various pathogens, including the presence of Vairimorpha spp. (Nosema spp.), quintefied Paenibacillus larvae, Acute Bee Paralysis Virus (ABPV), Black Queen Cell Virus (BQCV), Deformed Wing Virus (DWV), and Sacbrood Virus (SBV). However, our analysis did not detect Melissococcus plutonius, Crithidia mellificae, Lotmaria passim, and Aethina tumida. Samples of winter debris were also examined for the presence and quantification of the V. destructor mites, and their natural mite fall was observed in spring. Honeybee colonies were simultaneously infected by an average of four to six pathogens. Some observed honeybee colonies developed characteristic symptoms, while others did not survive the winter. Full article

This article systematically reviews controlled field trials of fumagillin dicyclohexylamine in honeybee colonies to determine whether fumagillin effectively controls nosema and whether it is beneficial to colonies. Fifty publications were found that described controlled field trials of fumagillin in honeybee colonies between 1952 and 2023. Fumagillin consistently reduced the prevalence and severity of nosema infections. Doses applied in recent studies were similar to or below those recommended historically. Furthermore, our study showed no negative effects on colony health. Improvements in colony survival, size, and honey production have been demonstrated frequently, though not consistently, in both historic and recent studies. Nevertheless, some practices are not optimal. Treatment decision thresholds based on the number of spores per bee are not well supported by evidence and may be no better than calendar-based prophylactic treatments. In addition, reasonable recommendations to employ quarantine and disinfection procedures together with fumagillin treatment do not appear to have been widely adopted. When used as stand-alone treatments, both the fall- and spring-label doses provide benefits but may be too low and short-term to ensure full control of the disease. Full article

Nosema bombycis is a representative species of Microsporidia, and is the pathogen that causes pebrine disease in silkworms. In the process of infection, the polar tube of N. bombycis is injected into the host cells. During proliferation, N. bombycis recruits the mitochondria of host cells. The general transcriptional corepressor Ssn6 contains six tetratricopeptide repeats (TPR) and undertakes various important functions. In this study, we isolated and characterized Nbssn6 of the microsporidium N. bombycis. The Nbssn6 gene contains a complete ORF of 1182 bp in length that encodes a 393 amino acid polypeptide. Indirect immunofluorescence assay showed that the Ssn6 protein was mainly distributed in the cytoplasm and nucleus at the proliferative phase of N. bombycis. We revealed the interaction of Nbssn6 with polar tube protein 2 (Nbptp2) and the transcriptional repressor for RNA polymerase II (Nbtrrp2) by Co-IP and yeast two-hybrid assays. Results from RNA interference further confirmed that the transcriptional level of Nbptp2 and Nbtrrp2 was regulated by Nbssn6. These results suggest that Nbssn6 impacts the infection and proliferation of N. bombycis via interacting with the polar tube protein and transcriptional repressor for RNA polymerase II. Full article

Mexico is a major honey producer, but not much information exists about the health status of honey bees (Apis mellifera L.) in the country. This study was conducted to determine the sanitary status of adult honey bees in Mexico’s five beekeeping regions. Samples from 369 apiaries were diagnosed to identify pathogens such as Varroa destructor, which was quantified, Acarapis woodi, Nosema spp., and five viruses. Colonies were also inspected for the presence of the small hive beetle (SHB), Aethina tumida. Varroa destructor was found in 83.5% of the apiaries, with the Pacific Coast region having the highest prevalence (>95%) and rates (4.5% ± 0.6). Acarapis woodi was detected in only one apiary from the Pacific Coast, whereas Nosema spp. were prevalent in 48.5% of the apiaries, with the highest and lowest frequencies in the Yucatan Peninsula and North regions (64.6% and 10.2%, respectively). For viruses, deformed wing virus (DWV) was detected in 26.1% of the apiaries, with the highest frequency in the Pacific Coast region (44.7%). Israeli acute paralysis virus (IAPV) was diagnosed in 3.2% of the samples and sacbrood bee virus (SBV) in 23.3% of them, with the highest frequency in the High Plateau region (36.4%). Chronic bee paralysis and Kashmir bee viruses were not detected. SHB prevalence was 25.2% nationwide, with the highest frequency in the Yucatan Peninsula (39.2%). This study shows that the most common parasites of adult honey bees in Mexico are V. destructor and Nosema spp., and that the most prevalent virus is DWV, whereas SHB is highly prevalent in the Yucatan Peninsula. This information could be useful to design disease control strategies for honey bee colonies in different regions of Mexico. Full article

The declining honeybee populations are a significant risk to the productivity and security of agriculture worldwide. Although there are many causes of these declines, parasites are a significant one. Disease glitches in honeybees have been identified in recent years and increasing attention has been paid to addressing the issue. Between 30% and 40% of all managed honeybee colonies in the USA have perished annually over the past few years. American foulbrood (AFB) and European foulbrood (EFB) have been reported as bacterial diseases, Nosema as a protozoan disease, and Chalkbrood and Stonebrood as fungal diseases. The study aims to compare the bacterial community related to the Nosema ceranae and Ascosphaera apis infection on the gut of the honeybee and compare it with the weakly active honeybees. The Nosema-infected honeybees contain the phyla Proteobacteria as the significantly dominant bacterial phyla, similar to the weakly active honeybees. In contrast, the Ascosphaera (Chalkbrood) infected honeybee contains large amounts of Firmicutes rather than Proteobacteria. Full article

Microsporidia are ubiquitous in the environment, infecting almost all invertebrates, vertebrates, and some protists. The microsporidian Nosema bombycis causes silkworms pébrine disease and leads to huge economic losses. Parasite secreted proteins play vital roles in pathogen–host interactions. Serine protease inhibitors (serpins), belonging to the largest and most broadly distributed protease inhibitor superfamily, are also found in Microsporidia. In this study, we characterized 19 serpins (NbSPNs) in N. bombycis; eight of them were predicted with signal peptides. All NbSPN proteins contain a typical conserved serpin (PF00079) domain. The comparative genomic analysis revealed that microsporidia serpins were only found in the genus Nosema. In addition to N. bombycis, a total of 34 serpins were identified in another six species of Nosema including N. antheraeae (11), N. granulosis (8), Nosema sp. YNPr (3), Nosema sp. PM-1 (3), N. apis (4), and N. ceranae (5). Serpin gene duplications in tandem obviously occurred in Nosema antheranae. Notably, the NbSPNs were phylogenetically clustered with serpins from the Chordopoxvirinae, the subfamily of Poxvirus. All 19 NbSPN transcripts were detected in the infected midgut and fat body, while 19 NbSPN genes except for NbSPN12 were found in the transcriptome of the infected silkworm embryonic cell line BmE-SWU1. Our work paves the way for further study of serpin function in microsporidia. Full article

Nosemosis type C is a dangerous and widespread disease of the adult European honey bee Apis mellifera and is caused by the spore-forming intracellular parasite Vairimorpha (Nosema) ceranae. The search for new ways of therapy for this disease is complicated due to the seasonal availability of V. ceranae-infected insects as well as the lack of a developed system for the pathogen’s cultivation. By carrying out trials which used different infectious dosages of the parasite, spore storage protocols, host age, and incubation temperatures, we present a simple, safe, and efficient method of V. ceranae propagation in artificially infected worker bees in the laboratory. The method is based on feeding the groups of adult worker bees with microsporidian spores and insect maintenance in plastic bottles at 33 °C. The source of the spores originated from the cadavers of infected insects from the previous round of cultivation, in which the infective spores persist for up to six months. An analysis of five independent cultivation rounds involving more than 2500 bees showed that the proposed protocol exploiting the dosage of one million spores per bee yielded over 60 million V. ceranae spores per bee, and most of the spore samples can be isolated from living insects. Full article

Nosema ceranae Fries, 1996, causes contagious fungal nosemosis disease in managed honey bees, Apis mellifera L. It is associated around the world with winter losses and colony collapse disorder. We used a laboratory in vivo screening assay to test curcumin, fenbendazole, nitrofurazone and ornidazole against N. ceranae in honey bees to identify novel compounds with anti-nosemosis activity compared to the commercially available medication Fumagilin-B^®. Over a 20-day period, Nosema-inoculated bees in Plexiglas cages were orally treated with subsequent dilutions of candidate compounds, or Fumagilin-B^® at the recommended dose, with three replicates per treatment. Outcomes indicated that fenbendazole suppressed Nosema spore proliferation, resulting in lower spore abundance in live bees (0.36 ± 1.18 million spores per bee) and dead bees (0.03 ± 0.25 million spores per bee), in comparison to Fumagilin-B^®-treated live bees (3.21 ± 2.19 million spores per bee) and dead bees (3.5 ± 0.6 million spores per bee). Our findings suggest that Fumagilin-B^® at the recommended dose suppressed Nosema. However, it was also likely responsible for killing Nosema-infected bees (24% mortality). Bees treated with fenbendazole experienced a greater survival probability (71%), followed by ornidazole (69%), compared to Nosema-infected non-treated control bees (20%). This research revealed that among screened compounds, fenbendazole, along with ornidazole, has potential effective antifungal activities against N. ceranae in a controlled laboratory environment. Full article

The honey bee is an important economic insect due to its role in pollinating many agricultural plants. Unfortunately, bees are susceptible to many pathogens, including pests, parasites, bacteria, and viruses, most of which exert a destructive impact on thousands of colonies. The occurrence of resistance to the therapeutic substances used against these organisms is rising, and the residue from these chemicals may accumulate in honey bee products, subsequently affecting the human health. There is current advice to avoid the use of antibiotics, antifungals, antivirals, and other drugs in bees, and therefore, it is necessary to develop alternative strategies for the treatment of bee diseases. In this context, the impact of nosema diseases (nosemosis) on bee health and the negative insults of existing drugs are discussed. Moreover, attempts to combat nosema through the use of alternative compounds, including essential oils, plant extracts, and microbes in vitro and in vivo, are documented. Full article

Honeybees are important pollinators, but they are continuously exposed to a variety of fungal and bacterial diseases. One of the various diseases affecting honeybees is nosemosis caused by microsporidia from the Nosema genus. Honeybees are mainly infected through consumption of infected food or faeces containing Nosema spp. spores. Nosemosis causes damage to the middle intestine epithelium, which leads to food absorption disorders and honeybee malnutrition. Fumagillin, i.e., the antibiotic used to treat nosemosis, was withdrawn in 2016 from EU countries. Therefore, researchers have been looking for compounds of both natural and synthetic origin to fight nosemosis. Such compounds should not have a negative impact on bees but is expected to inhibit the disease. Natural compounds tested against nosemosis include, e.g., essential oils (EOs), plant extracts, propolis, and bacterial metabolites, while synthetic substances tested as anti-nosemosis agents are represented by porphyrins, vitamins, antibiotics, phenolic, ascorbic acids, and others. This publication presents an 18-year overview of various studies of a number of natural and synthetic compounds used in the treatment and prevention of nosemosis cited in PubMed, GoogleScholar, and CrossRef. Full article

Beekeeping has yet to reach its full potential in terms of productivity in Turkey where it has a relatively large role in the economy. Poor colony health is suspected to be the reason for this, but comprehensive disease monitoring programs are lacking to support this notion. We sampled a total of 115 colonies across five different apiaries throughout the Marmara region of Turkey and screened for all of the major bee pathogens using PCR and RNA-seq methods. We found that Varroa mites are more prevalent in comparison to Nosema infections. The pathogens ABPV, DWV, KV, and VDV1 are near 100% prevalent and are the most abundant across all locations, which are known to be vectored by the Varroa mite. We therefore suspect that controlling Varroa mites will be key for improving bee health in Turkey moving forward. We also documented significant interactions between DWV, KV, and VDV1, which may explain how the more virulent strain of the virus becomes abundant. ABPV had a positive interaction with VDV1, thereby possibly facilitating this more virulent viral strain, but a negative interaction with Nosema ceranae. Therefore, these complex pathogen interactions should be taken into consideration in the future to improve bee health. Full article