Chen Y.P., Evans J.D., Murphy C., Gutell R., Zuker M., Gundersen-Rindal D., and Pettis J.S. (2009).
Morphological, Molecular, and Phylogenetic Characterization of Nosema cerenae, a Microsporidian Parasite Isolated from the European Honey Bee, Apis mellifera.
The Journal of Eukaryotic Microbiology, 56(2):142-147.
Genotoxicity of Eleusine indica (Nkim enang: Efik) was investigated in the Wister strain albino rat (Rattus novergicus). Nine (9) male and nine (9) female rats were randomly assigned to three (3) groups, of which two were exposed to the aqueous extract of E.indica ā Group A (control-no extract)), Group B (50 mg/kg BW of E. indica) and Group C (100 mg/kg BW of E. indica). This was administered to the rats by oral gavage for 14 days after which the peripheral blood from the tail tips were collected and assayed for the presence of micronuclei, following standard procedures. Proximate analysis and phytochemical screening of the herb extract was carried out. Results obtained showed that E. indica did not cause any significant (P > 0.05) increase in the incidence of micronucleated polychromatic erythrocytes in rat peripheral blood at any of the doses administered. The polychromatic: normochromatic erythrocyte (PCE: NCE) ratio was found to be in the range of 0.50 Ā± 0.11 to 0.55 Ā± 0.02. Also, the aqueous herb extract is rich in Carbohydrates (76.17%) and Tannins (21.76%). Mean body weights (MBW) of rats showed normal distribution throughout the duration of the investigation. The results of this study demonstrate that E. indica does not confer any genotoxicity in mammals. Further in-depth study on its efficacy is recommended.
In nature, wild animals live on large area and have consequently, a low genetic resistance against parasitic infections because of hoe exposure. When herds of these wild animals are kept in captivity in Zoological Gardens, the problem of parasite infection can aggravate and pose a serious threat to endangered species, occasionally causing sudden and unexpected local declines in abundance; unfortunately, there have been few detailed and comprehensive studies on the common parasitic infections, prevalence of the parasitic infections in the primates and the Health care management of the captive primates. The focus of the research work is on investigation of parasitic infection among primates in selected Zoological Gardens in Nigeria the researcher has the following objectives, The researcher adopted experimental method sample of fresh feaces were collected differently for 5 days and was examined at two different laboratory, that Ibadan and Jos. The Ibadan Zoological Garden twenty seven species of primates were examined, with only six infested with Trichuris Trichuria parasite; common in chimpanzee, mona monkey, Tantalus and white throated. In Jos Plateau Zoological Garden, twenty-four species of primates were examined only thirteen primates were infested with Trichuris Trichuria, F buskii, Eimeria, Ascaris Lumbricoides, Ā¬S. mansoni, Hetrophyes, Those infested, are Red patas, softy mongabey, Baboon, Mona Monkey, Tantalus, and Chimpanzee. Kano Zoological Garden, thirty species of primates were examined, only eleven were infested with Ascaris Lumbricoides, Eimeria, Trichuris Trichuria, Fasciola those infested are Baboon, Chimpanzee, Mona Monkey, Patas, Tantalus Monkeys. All the above examination of the faecal sample material, a direct wet smear was used to reveal the parasites; examined with a how power objective (10x).
this presentation deals with transgenic mosquitoes,strategies of vector control by transgenic mosquitoes, methods of transgenesis in mosquitoes, its drawbacks and its limitations
Prevalence Study of Infectious Bovine Keratoconjunctivitisin Dairy cattle und...iosrjce
Ā
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Genotoxicity of Eleusine indica (Nkim enang: Efik) was investigated in the Wister strain albino rat (Rattus novergicus). Nine (9) male and nine (9) female rats were randomly assigned to three (3) groups, of which two were exposed to the aqueous extract of E.indica ā Group A (control-no extract)), Group B (50 mg/kg BW of E. indica) and Group C (100 mg/kg BW of E. indica). This was administered to the rats by oral gavage for 14 days after which the peripheral blood from the tail tips were collected and assayed for the presence of micronuclei, following standard procedures. Proximate analysis and phytochemical screening of the herb extract was carried out. Results obtained showed that E. indica did not cause any significant (P > 0.05) increase in the incidence of micronucleated polychromatic erythrocytes in rat peripheral blood at any of the doses administered. The polychromatic: normochromatic erythrocyte (PCE: NCE) ratio was found to be in the range of 0.50 Ā± 0.11 to 0.55 Ā± 0.02. Also, the aqueous herb extract is rich in Carbohydrates (76.17%) and Tannins (21.76%). Mean body weights (MBW) of rats showed normal distribution throughout the duration of the investigation. The results of this study demonstrate that E. indica does not confer any genotoxicity in mammals. Further in-depth study on its efficacy is recommended.
In nature, wild animals live on large area and have consequently, a low genetic resistance against parasitic infections because of hoe exposure. When herds of these wild animals are kept in captivity in Zoological Gardens, the problem of parasite infection can aggravate and pose a serious threat to endangered species, occasionally causing sudden and unexpected local declines in abundance; unfortunately, there have been few detailed and comprehensive studies on the common parasitic infections, prevalence of the parasitic infections in the primates and the Health care management of the captive primates. The focus of the research work is on investigation of parasitic infection among primates in selected Zoological Gardens in Nigeria the researcher has the following objectives, The researcher adopted experimental method sample of fresh feaces were collected differently for 5 days and was examined at two different laboratory, that Ibadan and Jos. The Ibadan Zoological Garden twenty seven species of primates were examined, with only six infested with Trichuris Trichuria parasite; common in chimpanzee, mona monkey, Tantalus and white throated. In Jos Plateau Zoological Garden, twenty-four species of primates were examined only thirteen primates were infested with Trichuris Trichuria, F buskii, Eimeria, Ascaris Lumbricoides, Ā¬S. mansoni, Hetrophyes, Those infested, are Red patas, softy mongabey, Baboon, Mona Monkey, Tantalus, and Chimpanzee. Kano Zoological Garden, thirty species of primates were examined, only eleven were infested with Ascaris Lumbricoides, Eimeria, Trichuris Trichuria, Fasciola those infested are Baboon, Chimpanzee, Mona Monkey, Patas, Tantalus Monkeys. All the above examination of the faecal sample material, a direct wet smear was used to reveal the parasites; examined with a how power objective (10x).
this presentation deals with transgenic mosquitoes,strategies of vector control by transgenic mosquitoes, methods of transgenesis in mosquitoes, its drawbacks and its limitations
Prevalence Study of Infectious Bovine Keratoconjunctivitisin Dairy cattle und...iosrjce
Ā
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggpla...ijtsrd
Ā
Eggplant is prone to attack by several pests including bacteria, fungi, nematodes and insects. In this study, we have analyzed phylotype of bacterial wilt Ralstonia solanacearum infection in eggplant plants collected from Bhubaneswar Orissa in India. Bacterial wilt symptomatic five plant samples were collected from brinjal field in Bhubaneswar in 2016. The samples were macerated in sterile distilled water and grown on Kelman's triphenyltetrazolium chloride TZC agar media. Total genomic DNA of the bacterium were extracted and subjected to PCR amplification using the R. solanacearum specific universal primer pair 759 760. An expected single 280 bp fragment amplified in all the samples confirmed the identity of these as Ralstonia. To reconfirmed isolate of bacterium, the amplicon was sequenced in sequencer. In NCBI blast, the nucleotide sequence was 100 similar with Ralstonia solanacearum strain RS lpxC DOB 1 AB910593 and the sequence was submitted in NCBI database under Acc. No. KY393266. To determined phylotype of strain used specific multiplex PCR with phylotype specific primers Nmult 21F1 2, Nmult 22InF, Nmult 23AF, Nmult 22RR revealed that all the five infected samples belonged to phylotype I as a 144 bp amplicon were observed in agarose gel. On the basis of above finding concluded that the bacterial wilt infected eggplant collected from Bhubaneswar was Ralostonia solanacearum, Phylotype I. Rakesh Kumar | Ramachandran, E. | Koteshwar Yadav "Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggplants in Orissa in India" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21580.pdf
Molecular Identification of Bulinus Species in Ogun State, South-West Nigeria...AI Publications
Ā
The study considers the distribution of a small sample of 100 Bulinus snails, across 8 localities within Ogun State, Nigerian. Snails were identified using a molecular method of fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinustruncatus while only one was Bulinusglobosus. The use of Rsa1 restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 23% of snails were infected with schistosome
Introduction, importance and general characters of fungi, bacteria, fastidiou...Pankaj Thakur
Ā
I've prepared this wonderful eye catch presentation to explain you about these complex biology not in bulk but in points the points which holds power...
Pigeon Excreta A Potential Source of Cryptococcus Neoformans and their Antifu...ijtsrd
Ā
Globally the pigeon droppings are a known ecologic niche for cosmopolitan pathogenic yeast Cryptococcus neoformans, an etiological agent of deadly disease cryptococcosis. In this prospective study between 2015 2017, we analyzed the isolation of C. neoformans strains from a total of 305 pigeon excreta samples of caged pigeons with a pH of 6 8, from different sites of Central India. NCCLS broth microdilution methodology was employed on the isolated strains against amphotericin B, fluconazole, itraconazole, ketoconazole, and voriconazole. C. neoformans were found positive from fifty five dry guano feces. Maximum positive samples found for the pathogens were from caged pigeon excreta collected from the 12 different sites in city Jabalpur 23 46 , 9 18 from four sites katni, followed by 3 sites from each city Betul 8 16 , Satna 6 12 and Rewa 4 .08 . The highest frequency of C. neoformans was recorded from site 2 60 , followed by site 24 37.5 , site 17 27.27 , whereas site 3, 6, 10, 15 and 19 found negative for pathogenic yeast. the present study of antifungal susceptibility profile for C. neoformans revealed resistance against ketoconazole 25.5 and fluconazole 8.5 . The highest susceptibility was observed for amphotericin B 100 followed by voriconazole 97.9 and itraconazole 78.7 No resistance was found for polyene drug amphotericin B. Fluconazole 46.8 and ketoconazole 36.2 . This data of prevalence and colonization of this pathogen suggests that the dry excreta provides a more favorable environment for growth inside the cages and is more concerned with health hazards of the humans in proximity and further comprehensive study is required to reinforce the antifungal spectrum for the prudent therapy of cryptococcosis. Richa Gumasta | Shankar Mohan Singh | Ravi Prakash Mishra | Shesh Rao Nawange | Abhijeet Garg | Anuranjan Singh Rathore "Pigeon Excreta: A Potential Source of Cryptococcus Neoformans and their Antifungal Susceptibility Profile" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-5 , August 2019, URL: https://www.ijtsrd.com/papers/ijtsrd25250.pdfPaper URL: https://www.ijtsrd.com/biological-science/biotechnology/25250/pigeon-excreta-a-potential-source-of-cryptococcus-neoformans-and-their-antifungal-susceptibility-profile/richa-gumasta
The Sterile Insect Technique, best known by its acronym SIT and also identified as the Sterile Insect Release Method (SIRM), is a biologically-based method for the management of key insect pests of agricultural and medical/veterinary importance. In the FAO glossary, the Sterile Insect Technique is defined as "a method of pest control using area-wide inundative releases of sterile insects to reduce reproduction in a field population of the same species". It is therefore a type of "birth control" in which wild female insects of the pest population do not reproduce when they are inseminated by released, radiation-sterilized males. Sterilization is induced through the effects of irradiation on the reproductive cells of the insects. SIT does not involve the release of insects modified through transgenic (genetic engineering) processes. In this type of autocidal control, sequential releases of the sterilized insects in adequate sterile to wild male overflooding ratio's lead to a reduction in pest population numbers
Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggpla...ijtsrd
Ā
Eggplant is prone to attack by several pests including bacteria, fungi, nematodes and insects. In this study, we have analyzed phylotype of bacterial wilt Ralstonia solanacearum infection in eggplant plants collected from Bhubaneswar Orissa in India. Bacterial wilt symptomatic five plant samples were collected from brinjal field in Bhubaneswar in 2016. The samples were macerated in sterile distilled water and grown on Kelman's triphenyltetrazolium chloride TZC agar media. Total genomic DNA of the bacterium were extracted and subjected to PCR amplification using the R. solanacearum specific universal primer pair 759 760. An expected single 280 bp fragment amplified in all the samples confirmed the identity of these as Ralstonia. To reconfirmed isolate of bacterium, the amplicon was sequenced in sequencer. In NCBI blast, the nucleotide sequence was 100 similar with Ralstonia solanacearum strain RS lpxC DOB 1 AB910593 and the sequence was submitted in NCBI database under Acc. No. KY393266. To determined phylotype of strain used specific multiplex PCR with phylotype specific primers Nmult 21F1 2, Nmult 22InF, Nmult 23AF, Nmult 22RR revealed that all the five infected samples belonged to phylotype I as a 144 bp amplicon were observed in agarose gel. On the basis of above finding concluded that the bacterial wilt infected eggplant collected from Bhubaneswar was Ralostonia solanacearum, Phylotype I. Rakesh Kumar | Ramachandran, E. | Koteshwar Yadav "Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggplants in Orissa in India" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21580.pdf
Molecular Identification of Bulinus Species in Ogun State, South-West Nigeria...AI Publications
Ā
The study considers the distribution of a small sample of 100 Bulinus snails, across 8 localities within Ogun State, Nigerian. Snails were identified using a molecular method of fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinustruncatus while only one was Bulinusglobosus. The use of Rsa1 restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 23% of snails were infected with schistosome
Introduction, importance and general characters of fungi, bacteria, fastidiou...Pankaj Thakur
Ā
I've prepared this wonderful eye catch presentation to explain you about these complex biology not in bulk but in points the points which holds power...
Pigeon Excreta A Potential Source of Cryptococcus Neoformans and their Antifu...ijtsrd
Ā
Globally the pigeon droppings are a known ecologic niche for cosmopolitan pathogenic yeast Cryptococcus neoformans, an etiological agent of deadly disease cryptococcosis. In this prospective study between 2015 2017, we analyzed the isolation of C. neoformans strains from a total of 305 pigeon excreta samples of caged pigeons with a pH of 6 8, from different sites of Central India. NCCLS broth microdilution methodology was employed on the isolated strains against amphotericin B, fluconazole, itraconazole, ketoconazole, and voriconazole. C. neoformans were found positive from fifty five dry guano feces. Maximum positive samples found for the pathogens were from caged pigeon excreta collected from the 12 different sites in city Jabalpur 23 46 , 9 18 from four sites katni, followed by 3 sites from each city Betul 8 16 , Satna 6 12 and Rewa 4 .08 . The highest frequency of C. neoformans was recorded from site 2 60 , followed by site 24 37.5 , site 17 27.27 , whereas site 3, 6, 10, 15 and 19 found negative for pathogenic yeast. the present study of antifungal susceptibility profile for C. neoformans revealed resistance against ketoconazole 25.5 and fluconazole 8.5 . The highest susceptibility was observed for amphotericin B 100 followed by voriconazole 97.9 and itraconazole 78.7 No resistance was found for polyene drug amphotericin B. Fluconazole 46.8 and ketoconazole 36.2 . This data of prevalence and colonization of this pathogen suggests that the dry excreta provides a more favorable environment for growth inside the cages and is more concerned with health hazards of the humans in proximity and further comprehensive study is required to reinforce the antifungal spectrum for the prudent therapy of cryptococcosis. Richa Gumasta | Shankar Mohan Singh | Ravi Prakash Mishra | Shesh Rao Nawange | Abhijeet Garg | Anuranjan Singh Rathore "Pigeon Excreta: A Potential Source of Cryptococcus Neoformans and their Antifungal Susceptibility Profile" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-5 , August 2019, URL: https://www.ijtsrd.com/papers/ijtsrd25250.pdfPaper URL: https://www.ijtsrd.com/biological-science/biotechnology/25250/pigeon-excreta-a-potential-source-of-cryptococcus-neoformans-and-their-antifungal-susceptibility-profile/richa-gumasta
The Sterile Insect Technique, best known by its acronym SIT and also identified as the Sterile Insect Release Method (SIRM), is a biologically-based method for the management of key insect pests of agricultural and medical/veterinary importance. In the FAO glossary, the Sterile Insect Technique is defined as "a method of pest control using area-wide inundative releases of sterile insects to reduce reproduction in a field population of the same species". It is therefore a type of "birth control" in which wild female insects of the pest population do not reproduce when they are inseminated by released, radiation-sterilized males. Sterilization is induced through the effects of irradiation on the reproductive cells of the insects. SIT does not involve the release of insects modified through transgenic (genetic engineering) processes. In this type of autocidal control, sequential releases of the sterilized insects in adequate sterile to wild male overflooding ratio's lead to a reduction in pest population numbers
Ultrastructural Study of two Parasites Infecting Domesticated Turkey Meleagri...IOSRJPBS
Ā
This work provides a detailed systematic morphology by optic and Scanning electron microscopy of two parasites Raillietina echinobothriida Megnin, 1880 and Spirora meleagaris n. sp. infecting domesticated turkey. The present study includes some important characters that not recorded in previous description. SEM revealed that the tegument of the first cestode exhibits, filamentous, microtriches and sensory papillae densely covered the tegument of entire body, rostellum armed with two rows of hummer-shaped hooks and provide by 16 ā 20 rows of small, rose thorn-shaped accessory spines. In addition, the present studies have observed a number of taxonomic features in Spirora meleagaris n. sp. that differ from those mentioned in the same genus, mouth circular, bounded by a cuticular three circles plates, five pairs of cephalic papillae, an inner circle of two pairs situated on the wall of the buccal cavity, one pair of larges submedian amphids, and an outer circle of two pairs papillae. Buccal cavity supported by four chitinious cusped molar teeth anteriorly directed .Vulva near the end of the first third of the body, vulvular lips prominent. The male has unique rose like shaped pedunculated and unarranged numerous distributed sessile cervical papillae at the second third of the body that are distinguishable from other spirorid.
Prevelance of Lyperosomum longicauda Rudolphi, 1809 (Dicrocoeliioidae: Tremat...Innspub Net
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The present findings are related to reporting of the helminth parasitic infection in the Jungle babbler, at District: Naushahro Feroze. Host species were investigated from the month of June to August, 2018. These birds are non-migratory, former friendly, earth-colored siblings inhabit but internal visceral organs consisting intensity of parasites. Total (n=16) of T. striata were captured and dissected on a weekly basis under laboratory conditions at the Department of Zoology, SALU-Khairpur. All were found with the helminth population of digenean trematode but high prevalence was found in the month of June followed by other months. During surgical examination (n=44) specimens were recovered in the gall bladder of the host, morphologically having tapered ends at terminal body point, forebody is shorter than the hind body, protrusible rounded oral suckers but ventral suckers are rounded, maximum width at the post-acetabular region, oval-shaped pharynx, short esophagus, diverticular caeca, median-shaped ovary, and oblique testes, un-equal bands of lateral Stellaria and dark brown colored eggs. These features of the worms resemble already identified as; L. longicauda hence; identified as such. This species of fluke was first time recovered from the present host and the result of the present study revealed that it is a new host record from upper Sindh.
Introduction to endophytes and their application to develop commercial productsPrograma TF Innova
Ā
Ponencia: Introduction to endophytes and their application to develop commercial products
Autor: Dr. Gary Strobel
Evento TF Innova: Workshop Biotechnology "Isolation and identification of endophytic fungi from vascular plants"
PREVALENCE AND CHARACTERIZATION OF VIRULENCE PROPERTIES OF PSEUDOMONAS AERUGI...SUS GROUP OF INSTITUTIONS
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Pseudomonas aeruginosa is the epitome of an opportunistic pathogen of humans that cause urinary tract infections, respiratory system infection, particularly in victim of severe burns, cancer and AIDS patient who are immunocompromised. Most Pseudomonas infections are both invasive and toxigenic. The particular bacterial determinants of virulence mediate different stages of infection and are ultimately responsible for the characteristic syndromes that accompany the disease. In the present study P. aeruginosa was found to be more prevalent in burn patients (100%) followed by urinary tract infection samples (71%), sputum samples (66%) and wound samples (59%). 85% isolates recovered from clinical samples were mucoid. A total of 35% isolates were strong siderophore producers, 19% isolates were strong protease producers while 52% were strong phospholipase producers. Isolates from burns, sputum and environment sample were strong rhamnolipid producers. Elevated level of hemolysin production was observed in burn, urine and wound isolates. The prominence of haemagglutination ability in environmental isolates followed by burns isolates provided evidence for its being a nosocomial pathogen. The association between virulence determinants and disease can indicate the precise role played by the determinant in estabilishing the disease. Isolates were maximally sensitive towards ļ¢ lactam antibiotics.
Evolution of North American MicruracarusRachel Shoop
Ā
My research focuses on the evolution of North American water mites in the genus Arrenurus, Subgenus Micruracarus. In this presentation, I discuss why I chose to study these little known critters, and present some preliminary findings. Please contact me for more info.
palynotaxa and parasitic loads of nigerian currency potential sources of mic...INFOGAIN PUBLICATION
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Currency notes are handled by a large number of people under a variety of personal and environmental conditions. A total of ninety six samples of one hundred naira denomination of Nigerian notes were procured from seven Local Government Areas (LGA) of Ebonyi State, Nigeria. The aim of the study was to determine the palynotaxa and parasitic load prevalent on currency notes. The leachates of currency notes were obtained and subjected to acetolysis and examined microscopically. Twenty six fungal spores type were recorded and were highly dominated by spores of Libertelli spp., Botrytis spp. and Spadicoides spp. Pollen achieved 54 % of the total bio-particles, whereas fungal spores and parasitic worms achieved 35.2 % and 10.60 %, respectively. The presence and relative abundance of these palynotaxa and parasites in currency notes affirms their propensity to spread vectors of diseases.
The Effect of Dried Leaves Extract of Hyptis suaveolens on Various Stages of ...iosrjce
Ā
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Gutell 123.app environ micro_2013_79_1803Robin Gutell
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McFrederick Q.S., Cannone J.J., Gutell R.R., Kellner K., Plowes R.M., and Mueller U.G. (2013).
Specificity between Lactobacilli and Hymenopteran Hosts is the Exception Rather than the Rule.
Applied and Environmental Microbiology, 79:1803-1812.
Gutell R.R. (2013).
Comparative Analysis of the Higher-Order Structure of RNA.
in: Biophysics of RNA Folding. Volume editor: Rick Russell. Series title: Biophysics for the Life Sciences. Series editors: Norma Allewell, Ivan Rayment, Bertrand Garcia-Moreno, Jonathan Dinman, and Michael McCarthy. pp. 11-22. Publisher: Springer, New York, NY.
Gardner D.P., Xu W., Miranker D.P., Ozer S., Cannone J.J., and Gutell R.R. (2012).
An Accurate Scalable Template-based Alignment Algorithm.
Proceedings of 2012 IEEE International Conference on Bioinformatics and Biomedicine (BIBM 2012), Philadelphia, PA. October 4-7, 2012. IEEE Computer Society, Washington, DC, USA. pp. 237-243.
Lee J.C. and Gutell R.R. (2012).
A Comparison of the Crystal Structures of the Eukaryotic and Bacterial SSU Ribosomal RNAs Reveals Common Structural Features in the Hypervariable Regions.
PLoS ONE, 7(5):e38203.
Gardner D.P., Ren P., Ozer S., and Gutell R.R. (2011).
Statistical Potentials for Hairpin and Internal Loops Improve the Accuracy of the Predicted RNA Structure.
Journal of Molecular Biology, 413(2):473-483.2011. pp 15-22.
Ozer S., Doshi K.J., Xu W., and Gutell R.R. (2011).
rCAD: A Novel Database Schema for the Comparative Analysis of RNA.
7th IEEE International Conference on e-Science, Stockholm, Sweden. December 5-8, 2011. pp 15-22.
Jiang Y., Xu W., Thompson L.P., Gutell R., and Miranker D. (2011).
R-PASS: A Fast Structure-based RNA Sequence Alignment Algorithm.
Proceedings of 2011 IEEE International Conference on Bioinformatics and Biomedicine (BIBM 2011), Atlanta, GA. November 12-15, 2011. IEEE Computer Society, Washington, DC, USA. pp. 618-622.
Xu W., Wongsa A., Lee J., Shang L., Cannone J.J., and Gutell R.R. (2011).
RNA2DMap: A Visual Exploration Tool of the Information in RNA's Higher-Order Structure.
Proceedings of 2011 IEEE International Conference on Bioinformatics and Biomedicine (BIBM 2011), Atlanta, GA. November 12-15, 2011. IEEE Computer Society, Washington, DC, USA. pp. 613-617.
Muralidhara C., Gross A.M., Gutell R.R., and Alter O. (2011).
Tensor Decomposition Reveals Concurrent Evolutionary Convergences and Divergences and Correlations with Structural Motifs in Ribosomal RNA.
PLoS ONE, 6(4):e18768.
Xia Z., Gardner D.P., Gutell R.R., and Ren P. (2010).
Coarse-Grained Model for Simulation of RNA Three-Dimensional Structures.
The Journal of Physical Chemistry B, 114(42):13497-13506.
Milbury C.A., Lee J.C., Cannone J.J., Gaffney P.M., and Gutell R.R. (2010).
Fragmentation of the Large Subunit Ribosomal RNA Gene in Oyster Mitochondrial Genomes.
BMC Genomics, 11(1):485.
Mueller U.G., Ishak H., Lee J.C., Sen R., and Gutell R.R. (2010).
Placement of attine ant-associated Pseudonocardia in a global phylogeny (Pseudonocardiaceae, Actinomycetales): a test of two symbiont-association models.
Antonie van Leeuwenhoek International Journal of General and Molecular Microbiology, 98(2):195-212.
Theriot E.C., Cannone J.J., Gutell R.R., and Alverson A.J. (2009).
The limits of nuclear encoded SSU rDNA for resolving the diatom phylogeny.
European Journal of Phycology, 44(3):277-290.
Wu J.C., Gardner D.P., Ozer S., Gutell R.R. and Ren P. (2009).
Correlation of RNA Secondary Structure Statistics with Thermodynamic Stability and Applications to Folding.
Journal of Molecular Biology, 391(4):769-783.
Xu W., Ozer S., and Gutell R.R. (2009).
Covariant Evolutionary Event Analysis for Base Interaction Prediction Using a Relational Database Management System for RNA.
21st International Conference on Scientific and Statistical Database Management. June 2-4, 2009. Springer-Verlag. pp. 200-216.
Maddison D.R., Moore W., Baker M.D., Ellis T.M., Ober K.A., Cannone J.J., and Gutell R.R. (2009).
Monophyly of terrestrial adephagan beetles as indicated by three nuclear genes (Coleoptera: Carabidae and Trachypachidae).
Zoologica Scripta, 38(1):43-62.
GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
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Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
Climate Impact of Software Testing at Nordic Testing DaysKari Kakkonen
Ā
My slides at Nordic Testing Days 6.6.2024
Climate impact / sustainability of software testing discussed on the talk. ICT and testing must carry their part of global responsibility to help with the climat warming. We can minimize the carbon footprint but we can also have a carbon handprint, a positive impact on the climate. Quality characteristics can be added with sustainability, and then measured continuously. Test environments can be used less, and in smaller scale and on demand. Test techniques can be used in optimizing or minimizing number of tests. Test automation can be used to speed up testing.
Smart TV Buyer Insights Survey 2024 by 91mobiles.pdf91mobiles
Ā
91mobiles recently conducted a Smart TV Buyer Insights Survey in which we asked over 3,000 respondents about the TV they own, aspects they look at on a new TV, and their TV buying preferences.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Ā
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
PHP Frameworks: I want to break free (IPC Berlin 2024)Ralf Eggert
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In this presentation, we examine the challenges and limitations of relying too heavily on PHP frameworks in web development. We discuss the history of PHP and its frameworks to understand how this dependence has evolved. The focus will be on providing concrete tips and strategies to reduce reliance on these frameworks, based on real-world examples and practical considerations. The goal is to equip developers with the skills and knowledge to create more flexible and future-proof web applications. We'll explore the importance of maintaining autonomy in a rapidly changing tech landscape and how to make informed decisions in PHP development.
This talk is aimed at encouraging a more independent approach to using PHP frameworks, moving towards a more flexible and future-proof approach to PHP development.
Welcome to the first live UiPath Community Day Dubai! Join us for this unique occasion to meet our local and global UiPath Community and leaders. You will get a full view of the MEA region's automation landscape and the AI Powered automation technology capabilities of UiPath. Also, hosted by our local partners Marc Ellis, you will enjoy a half-day packed with industry insights and automation peers networking.
š Curious on our agenda? Wait no more!
10:00 Welcome note - UiPath Community in Dubai
Lovely Sinha, UiPath Community Chapter Leader, UiPath MVPx3, Hyper-automation Consultant, First Abu Dhabi Bank
10:20 A UiPath cross-region MEA overview
Ashraf El Zarka, VP and Managing Director MEA, UiPath
10:35: Customer Success Journey
Deepthi Deepak, Head of Intelligent Automation CoE, First Abu Dhabi Bank
11:15 The UiPath approach to GenAI with our three principles: improve accuracy, supercharge productivity, and automate more
Boris Krumrey, Global VP, Automation Innovation, UiPath
12:15 To discover how Marc Ellis leverages tech-driven solutions in recruitment and managed services.
Brendan Lingam, Director of Sales and Business Development, Marc Ellis
In his public lecture, Christian Timmerer provides insights into the fascinating history of video streaming, starting from its humble beginnings before YouTube to the groundbreaking technologies that now dominate platforms like Netflix and ORF ON. Timmerer also presents provocative contributions of his own that have significantly influenced the industry. He concludes by looking at future challenges and invites the audience to join in a discussion.
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
Ā
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
The Metaverse and AI: how can decision-makers harness the Metaverse for their...Jen Stirrup
Ā
The Metaverse is popularized in science fiction, and now it is becoming closer to being a part of our daily lives through the use of social media and shopping companies. How can businesses survive in a world where Artificial Intelligence is becoming the present as well as the future of technology, and how does the Metaverse fit into business strategy when futurist ideas are developing into reality at accelerated rates? How do we do this when our data isn't up to scratch? How can we move towards success with our data so we are set up for the Metaverse when it arrives?
How can you help your company evolve, adapt, and succeed using Artificial Intelligence and the Metaverse to stay ahead of the competition? What are the potential issues, complications, and benefits that these technologies could bring to us and our organizations? In this session, Jen Stirrup will explain how to start thinking about these technologies as an organisation.
Epistemic Interaction - tuning interfaces to provide information for AI supportAlan Dix
Ā
Paper presented at SYNERGY workshop at AVI 2024, Genoa, Italy. 3rd June 2024
https://alandix.com/academic/papers/synergy2024-epistemic/
As machine learning integrates deeper into human-computer interactions, the concept of epistemic interaction emerges, aiming to refine these interactions to enhance system adaptability. This approach encourages minor, intentional adjustments in user behaviour to enrich the data available for system learning. This paper introduces epistemic interaction within the context of human-system communication, illustrating how deliberate interaction design can improve system understanding and adaptation. Through concrete examples, we demonstrate the potential of epistemic interaction to significantly advance human-computer interaction by leveraging intuitive human communication strategies to inform system design and functionality, offering a novel pathway for enriching user-system engagements.
Transcript: Selling digital books in 2024: Insights from industry leaders - T...BookNet Canada
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The publishing industry has been selling digital audiobooks and ebooks for over a decade and has found its groove. Whatās changed? What has stayed the same? Where do we go from here? Join a group of leading sales peers from across the industry for a conversation about the lessons learned since the popularization of digital books, best practices, digital book supply chain management, and more.
Link to video recording: https://bnctechforum.ca/sessions/selling-digital-books-in-2024-insights-from-industry-leaders/
Presented by BookNet Canada on May 28, 2024, with support from the Department of Canadian Heritage.
Removing Uninteresting Bytes in Software FuzzingAftab Hussain
Ā
Imagine a world where software fuzzing, the process of mutating bytes in test seeds to uncover hidden and erroneous program behaviors, becomes faster and more effective. A lot depends on the initial seeds, which can significantly dictate the trajectory of a fuzzing campaign, particularly in terms of how long it takes to uncover interesting behaviour in your code. We introduce DIAR, a technique designed to speedup fuzzing campaigns by pinpointing and eliminating those uninteresting bytes in the seeds. Picture this: instead of wasting valuable resources on meaningless mutations in large, bloated seeds, DIAR removes the unnecessary bytes, streamlining the entire process.
In this work, we equipped AFL, a popular fuzzer, with DIAR and examined two critical Linux libraries -- Libxml's xmllint, a tool for parsing xml documents, and Binutil's readelf, an essential debugging and security analysis command-line tool used to display detailed information about ELF (Executable and Linkable Format). Our preliminary results show that AFL+DIAR does not only discover new paths more quickly but also achieves higher coverage overall. This work thus showcases how starting with lean and optimized seeds can lead to faster, more comprehensive fuzzing campaigns -- and DIAR helps you find such seeds.
- These are slides of the talk given at IEEE International Conference on Software Testing Verification and Validation Workshop, ICSTW 2022.
1. Morphological, Molecular, and Phylogenetic Characterization of Nosema ceranae,
a Microsporidian Parasite Isolated from the European Honey Bee, Apis mellifera1
YANPING P. CHEN,a
JAY D. EVANS,a
CHARLES MURPHY,b
ROBIN GUTELL,c
MICHAEL ZUKER,d
DAWN GUNDENSEN-RINDALe
and JEFF S. PETTISa
a
USDA-ARS, Bee Research Laboratory, Beltsville, Maryland 20705, and
b
USDA-ARS, Soybean Genomic & Improvement Laboratory, Beltsville, Maryland 20705, and
c
Institute for Cellular and Molecular Biology and Section of Integrative Biology, University of Texas, Austin, Texas 78712, and
d
Rensselaer Polytechnic Institute, New York 12180, and
e
USDA-ARS, Invasive Insect Biocontrol and Behavior Laboratory, Beltsville, Maryland 20705
ABSTRACT. Nosema ceranae, a microsporidian parasite originally described from Apis cerana, has been found to infect Apis melllifera
and is highly pathogenic to its new host. In the present study, data on the ultrastructure of N. ceranae, presence of N. ceranae-speciļ¬c
nucleic acid in host tissues, and phylogenetic relationships with other microsporidia species are described. The ultrastructural features
indicate that N. ceranae possesses all of the characteristics of the genus Nosema. Spores of N. ceranae measured approximately
4.4 Ć 2.2 mm on fresh smears. The number of coils of the polar ļ¬lament inside spores was 18ā21. Polymerase chain reaction (PCR) signals
speciļ¬c for N. ceranae were detected not only in the primary infection site, the midgut, but also in the tissues of hypopharyngeal glands,
salivary glands, Malpighian tubules, and fat body. The detection rate and intensity of PCR signals in the fat body were relatively low
compared with other examined tissues. Maximum parsimony analysis of the small subunit rRNA gene sequences showed that N. ceranae
appeared to be more closely related to the wasp parasite, Nosema vespula, than to N. apis, a parasite infecting the same host.
Key Words. Disease, microorganism, parasitism, phylogeny.
SINCE their ļ¬rst recognition as pathogens in silkworms by
NageĀØli (1857), microsporidia have been identiļ¬ed as the
sources of many infectious diseases in vertebrates and inverte-
brates, including human, ļ¬shes, and insects (Canning, Lom, and
Dykova 1986; Franzen and MuĀØller 2001; Sprague and Vavra
1977; Wasson and Peper 2000; Wittner and Weiss 1999). Of 143
genera and over 1,200 microsporidian species described (Witt-
ner and Weiss 1999), insects are frequently found to be their pri-
mary host. Most of the entomopathogenic microsporidia occur in
the genus Nosema, which has 4150 described species and infects
nearly all taxonomic orders of insect, especially the orders of Le-
pidoptera and Hymenoptera (Becnel and Andreadis 1999; Sprague
1978).
Nosemosis is a serious disease of adult honey bees, caused by
Nosema species. Honey bee colonies are frequently infected, and
all colony members, including adult worker bees, drones, and
queens, can be infected. Nosema infection occurs mostly through
ingestion of spores with food or water. The physical and chemical
conditions of the midgut trigger the germination of spores and the
vegetative stage of Nosema begins to grow and multiply inside
midgut cells. Bailey and Ball (1997) showed that 30ā50 million
spores could be found inside a beeās midgut within 2 wk after
initial infection. Eventually the spores pass out of the bee in its
feces, providing new sources of the infection through cleaning and
feeding activities in the colonies.
Nosema infections have signiļ¬cant negative impacts on honey
bees, causing dysentery, shortened life spans of honey bees, su-
persedure of infected queens, and decrease in colony size (Has-
sanein 1953; Malone, Giacon, and Newton 1995; Rinderer and
Sylvester 1978). Nosema ceranae and Nosema apis are two spe-
cies of Nosema that are reported to infect the European honey bee,
Apis mellifera. For years, nosemosis of the European honey bee
was exclusively attributed to N. apis. Nosema ceranae, a species
originally found in the Asian honey bee, Apis cerana (Fries et al.
1996), is now a common infection of European honey bees and
is highly pathogenic to its new host (Cox-Foster et al. 2007; Fries
et al. 2006; Higes, Martin, and Meana 2006; Higes et al. 2007;
Huang et al. 2007; Klee et al. 2007). Chen et al. (2008) demon-
strated that N. ceranae was transferred from A. cerana to A. mell-
ifera at least a decade ago and is now replacing N. apis as the
predominant microsporidian infection in A. mellifera of the U.S.
populations. Although widespread infection by N. ceranae in the
U.S. population of A. mellifera has been identiļ¬ed, many biolog-
ical features of this parasite in the host A. mellifera remain to be
elucidated. To remedy this deļ¬ciency, we describe key morpho-
logical features of N. ceranae based on light and electron micros-
copy, and we use PCR to determine the presence of N. ceranae-
speciļ¬c nucleic acid in different tissues of the infected hosts. By
comparing the sequences of small subunit rRNA (SSUrRNA)
genes of different microsporidia, we construct a phylogenetic tree
to determine the genetic relationship of N. ceranae with other
species of microsporidia infecting insects.
MATERIALS AND METHODS
Honey bee sample collection. Honey bees were collected
from colonies maintained in Beltsville, MD. The abdomens of
10 honey bees from each colony were ground up in 2 ml of sterile
distilled water. One drop of the homogenate was examined by a
light microscope for presence of Nosema spores. When spores of
Nosema were observed under the microscope, the remaining por-
tion of the homogenized abdomens was used for DNA extraction
and PCR assays to determine the species status of Nosema infec-
tion. Once the Nosema infection of bee colonies was identiļ¬ed,
additional adult bees were collected from those heavily infected
colonies and stored at Ć 20 1C for subsequent morphological and
molecular analyses.
Puriļ¬cation of Nosema spores. To obtain puriļ¬ed Nosema
spores, the alimentary tracts of honey bees from Nosema-infected
colonies were removed individually by grasping the sting with
forceps and gently pulling the alimentary tract away from the ab-
domen. The midgut was separated from the hindgut and 25 pieces
of midgut were crushed in 2 ml of sterile distilled water and ļ¬l-
Corresponding Author: Y. Chen, USDA-ARS, Bee Research Labo-
ratory, Building 476, BARC-East Beltsville, MD 20705āTelephone
number: 11 301 504 8749; FAX number: 11 301 504 8736; e-mail:
judy.chen@ars.usda.gov
1
Disclaimer: Mention of trade names or commercial products in this
article is solely for the purpose of providing speciļ¬c information and
does not imply recommendation or endorsement by the U.S. Department
of Agriculture.
142
J. Eukaryot. Microbiol., 56(2), 2009 pp. 142ā147
r 2009 The Author(s)
Journal compilation r 2009 by the International Society of Protistologists
DOI: 10.1111/j.1550-7408.2008.00374.x
2. tered through a Corning Netwell (Corning Incorporated Life Sci-
ence, Lowell, MA) insert (24 mm diam., 74 mm mesh size) to re-
move tissue debris. The ļ¬ltered suspension was centrifuged at
1,500 g for 5 min and the supernatant was discarded. The pellet
was resuspended in 1 ml of sterile water and overlayed very gently
on a discontinuous 25%, 50%, 75%, and 100% of Percoll (Sigma-
Aldrich, St. Louis, MO) gradient from top to the bottom and cen-
trifuged twice at 8,000 g for 20 min at 4 1C using a Beckman rotor
(SW 28) in a Beckman L8-70M ultracentrifuge (Beckman Instru-
ments, Inc., Palo Alto, CA) to collect spores having the same size,
shape, and density. The supernatant was discarded and the spore
pellet was resuspended in distilled sterile water and collected by
centrifugation. After a ļ¬nal centrifugation at 8,000 g for 10 min at
4 1C, the spore pellet was resuspended in distilled sterile water and
stored at 4 1C until used. Spore sizes were measured under an
Eclipse TE 300 light microscope (Nikon, Melville, NY) and pho-
tographed with a Nikon Digital Camera (DXM 1200).
Light and electron microscopy. Midguts of adult bees from a
Nosema-infected colony were dissected out as described above
and midgut tissue was ļ¬xed for 2 h at room temperature by im-
mersion in 3% (v/v) glutaraldehyde in 0.05 M sodium cacodylate
buffer, pH 7.0. After overnight incubation in a refrigerator at 4 1C,
tissues were washed in a sodium cacodylate buffer rinse, 6 times
over 1 h, postļ¬xed in 2% (w/v) osmium tetroxide in 0.05 M so-
dium cacodylate buffer, pH 7.0, for 2 h, dehydrated in ethanol and
imbedded in Spurrās low-viscosity embedding resin. One-
micrometer-thick sections were cut on a Reichert/AO Ultracut
microtome (Leica Inc., Deerļ¬eld, IL) with a Diatome (Hatļ¬eld,
PA) diamond knife. The sections for light microscopy were
mounted onto slides, stained with 0.5% (w/v) toluidine blue and
photographed with the same system for spores. Sections for elec-
tron microscopy were mounted onto 200-mesh Ni grids, stained
with 4% (w/v) uranyl acetate and 3% (w/v) lead citrate, and
viewed in a H-7000 Hitachi (Tokyo, Japan) microscope at 75 kV.
Tissue dissection. Fifteen adult worker bees collected from
Nosema-infected colonies were used for tissue dissection. Tissues
of alimentary canal, Malpighian tubules, muscle, fat body, hypo-
pharyngeal gland, and salivary gland were carefully separated
under a Zeiss SV11 Stereomicroscope (Thornwood, NY) and pho-
tographed with a Zeiss AxioCam digital camera. All tissues were
rinsed once with 1 Ć PBS and twice with nuclease-free water to
prevent possible contamination and then subjected to subsequent
molecular analysis to determine the presence of N. ceranae-spe-
ciļ¬c nucleic acid in tissues.
DNA isolation, PCR ampliļ¬cation, and DNA sequencing.
Dissected tissues were frozen in liquid nitrogen individually and
ground to a ļ¬ne powder using a mortar and pestle. The genomic
DNA was extracted using the DNAzol DNA puriļ¬cation kit (Inv-
itrogen, Carlsbad, CA) following the manufacturerās protocol.
Two pairs of primers speciļ¬c for N. apis (N-apis-F: 50
-
ccattgccggataagagagt-30
; N-apis-R: 50
-cacgcattgctgcatcattgac-30
)
and N. ceranae (N-ceranae-F: 50
-cggataaaagagtccgttacc-30
; N-
ceranae-R: 50
-tgagcagggttctagggat-30
) were used in the study as
described previously (Chen et al. 2008). The speciļ¬city of ampli-
ļ¬cation was conļ¬rmed by cloning the puriļ¬ed PCR fragments
from 1.5% low-melting-point agarose gel using Wizard PCR Prep
DNA Puriļ¬cation System (Promega, Madison, WI) in pCR 2.1
vector (Invitrogen), and sequencing the PCR fragments from both
directions using M13-forward and M13-reverse primers. The se-
quence data was analyzed using the BLAST server at the National
Center for Biotechnology Information.
Phylogenetic analysis. The 21 species of microsporidian
SSUrRNA sequences with the highest BLAST similarity score
against the complete sequence of the N. ceranae SSUrRNA were
retrieved from GenBank database. The hosts of microsporidian
species used for phylogenetic analysis were all insects from the
Orders Hymenoptera, Lepidoptera, and Coleoptera. Trachipleis-
tophora hominis infecting Homo sapiens was used as an outgroup
to root the phylogenetic tree. Sequences were aligned using Meg-
Align (DNASTAR Lasergene software program, Madison, WI)
and sequences that could not be aligned unambiguously at both
30
- and 50
-ends were truncated. The percentage identity and
divergence of sequences between equivalent microsporidian
SSUrRNA was generated by the MegAlign. Aligned sequences
of 20 microsporidia species and the outgroup were imported into
the phylogenetic analysis program PAUP 4.03 (Sinauer Associ-
ates, Sunderland, MA). Maximum Parsimony under a heuristic
search with random stepwise addition and TBR branch swapping
was used to construct the phylogenetic trees. Phylogenies were
assessed by a 1,000 bootstrap replication.
RESULTS
Nosema ceranae infection was found in adult bees of A. mell-
ifera collected in Beltsville, MD. When the abdomens of infected
bees were crushed in water, a large numbers of mature spores
were released, although most infected bees did not exhibit overt
behavior and morphological signs of infection. The samples ex-
amined in this study were exclusively N. ceranae-positive: none
of the PCR reactions using N. apis-speciļ¬c primers yielded any
product (data not shown).
Light microscopy revealed that fresh N. ceranae spores were
oval or rod shaped, varied in size with a length of 3.9ā5.3 mm
(mean Ć SE 5 4.4 Ć 0.41 mm) and a width of 2.0ā2.5 mm (mean-
SE 5 2.2 Ć 0.09 mm) (N 5 50) (Fig. 1). Observation of Nosema-
infected midgut tissue showed that mature spores not only accu-
mulate in midgut epithelial cells but also are released into the gut
lumen (Fig. 2, 3).
Ultrastructural studies showed that different developmental
stages, including meronts, sporonts, sporoblasts, and mature
spores, are found in the midgut epithelial cells. Meronts, the ear-
liest developmental stage, had two nuclei in diplokarytic arrange-
ment and were bound by a plasma membrane in direct contact
with host cytoplasm (Fig. 4). Sporonts were elongated and oval in
shape with dense cytoplasm and no discernible internal structures
(Fig. 5). Sporoblasts were generally smaller than sporonts with a
more clearly deļ¬ned cell wall and two nuclei (Fig. 4). Electron
micrographs of longitudinal sections of mature spores showed that
the spore wall consisted of a dense exospore, 48ā53 nm thick, and
a lucent layer endospore, and that the sporoplasm was enclosed by
Fig. 1. Nosema ceranae spores. Light micrograph of oval- to rod-
shaped spores of N. ceranae after Percoll puriļ¬cation. Scale bar 5 10 mM.
143CHEN ET AL.āCHARACTERIZATION OF NOSEMA CERANAE
3. a plasma membrane (Fig. 6). The anchoring disk was located in
the anterior pole of the spore. The lamellate polaroplast occupied
the anterior part of the spore adjacent to the anchoring disk (Fig.
6). A vacuole was located in the posterior end of the spore and not
prominent. Two nuclei in diplokaryotic arrangement were closely
apposed in the central region of the spore between the polaroplast
and the posterior vacuole and the polar ļ¬laments were arranged in
18ā21 isoļ¬lar coils in two rows (Fig. 7). When a spore had an
extruded polar tube, the posterior vacuole swelled and became
very prominent inside the spore (Fig. 8).
The PCR assays revealed that N. ceranae-speciļ¬c nucleic acid
was detected in 100% of the alimentary canals, Malpighian tu-
bules, and hypopharyngeal glands, in 87% of the salivary glands,
and in 20% of the fat bodies (N 5 15). No N. ceranae-speciļ¬c
PCR signal was detected in the muscle tissue examined (lane 5,
Fig. 9).
The complete DNA sequences of the rRNA gene is 4,475 bp.
The G1C content of the SSUrRNA cistron at positions 1ā1,259
was 36.46%. The internal transcribed spacer (ITS) region con-
sisted of a 39-bp sequence and was located between nucleotides
1260 and 1298. The DNA sequence of LSUrRNA, located at the
30
-end between nucleotides 1299 and 3828, contained 2,530 bp
and was 32.86% G1C.
The percent of SSUrRNA sequence identity revealed that N.
ceranae shared the highest degree of sequence identity (97.5%)
with N. vespula and was the most distantly related to Nosema
plutellae with 19.3% sequence divergence among all micros-
poridia included in this study. Our phylogenetic tree of 20 mi-
crosporidian taxa contains two distinct clades (Fig. 10). One clade
includes Vairimorpha imperfecta and some species of the āātrueāā
Nosema group, a group of lepidopteran Nosema species closely
related to Nosema bombycis (Baker et al. 1994). Nosema ceranae,
along with several non-lepidoteran Nosema species and true Nose-
ma species forms another clade (Fig. 10). Within this latter clade,
Fig. 2, 3. Cross-section of the midgut showing spores. 2. Spores ac-
cumulated in midgut lumen. 3. Epithelial cells of the midgut infected with
Nosema ceranae. Arrows indicate infected epithelial cells with tightly
packed parasites.
Fig. 4, 5. Epithelial cells infected with different developmental stages
of Nosema ceranae. The developmental stages include meront (M), spo-
ront (ST), sporoblast (SB), and mature spore (MS). MB, membrane of the
infected host cell; ES, empty shell of the hatched spore.
144 J. EUKARYOT. MICROBIOL., 56, NO. 2, MARCHāAPRIL 2009
4. N. ceranae is most closely related to N. vespula with 80% boot-
strap support, and was distantly related to N. apis.
DISCUSSION
The transfer of N. ceranae from its described original host,
A. cerana, to a possible novel host, A. mellifera, adds a new di-
mension to the biological and epidemiological aspects of this par-
asite. Experimental infection of A. mellifera by N. ceranae
conducted by Higes et al. (2007) clearly showed that this para-
site is highly pathogenic to its new host and poses a serious threat
to the beekeeping industry.
The morphological and molecular characterization of N. cer-
anae in Asian honey bees was conducted by Fries et al. (1996).
Later, Fries et al. (2006) reported the natural infection N. ceranae
in European honey bees. However, many morphological details of
spores such as types and sizes of spores in a dense spore puriļ¬-
cation and the morphology at the different developmental stages
of spores in midgut epithelium cells in naturally infected hosts
remained to be demonstrated. Our observation with light micros-
copy showed that spores of N. ceranae from European honey bees
are oval shaped and rather uniform in shape. The electron mi-
croscopy indicates that N. ceranae contains all of the ultrastruc-
tural characteristics of the genus Nosema (Larsson 1986):
diplokaryotic nuclei present in all developmental stages; a long
ļ¬exible polar ļ¬lament that appears in the mature spores; meronts,
the earliest stages in the life cycle of the parasite, which are in
direct contact with host cell cytoplasm; mature spores that are
bounded by a thickened wall consisting of electron-dense exo-
spore and electron-lucent endospore layers; and the thickness of
exospore is 48ā52 nm. The number of polar ļ¬lament coils is an
important taxonomic criterion to differentiate different species of
Nosema (Burges, Canning, and Hulls 1974). The number of coils
of polar ļ¬lament inside N. ceranae spores measured by us was
18ā21, overlapping with the range of 20ā23 coils reported by
Fries et al. (1996), which is much smaller than the 430 coils re-
corded for N. apis (Fries 1989; Liu 1984).
Although not all examined tissues showed visible signs of
pathological changes, PCR assay followed by sequencing analy-
sis showed that N. ceranae-speciļ¬c PCR signals are not restricted
to the midgut tissue but spread to other tissues, including the
Malpighian tubules, hypopharyngeal glands, salivary glands, and
fat bodies. The presence of the signal suggests that these tissues
may be infected, as was determined microscopically for Nosema
bombi in a bumble bee (Fries et al. 2001). However, microscopic
studies of N. ceranae in A. mellifera tissues, which would verify
the infections, remain to be conducted. The detection of N. cer-
anae-speciļ¬c PCR signals in both hypopharyngeal and salivary
glands suggests that royal jelly, the secretion of hypopharyngeal
and salivary glands of worker bees used to feed the queen and
larvae, could be another vehicle for horizontal fecalāoral and
food-borne transmission of the parasite in the bee colonies. A
weak PCR signal speciļ¬c for N. ceranae detected in the fat body
tissue suggests a low parasite load, arguing that fat body tissue is
not a primary target for N. ceranae infection even though fat-body
tissues are one of the primary sites for microsporidian infection.
Infection of fat bodies causes formation of whitish cysts and the
infected gut becomes swollen and whitish as a result of impaired
fat metabolism in many other insects (Sokolova et al. 2006).
Although honey bee colonies with reduced longevity, de-
creased population size, higher autumn/winter colony loss, and/
or reduced honey production are often reported to be associated
with the presence of N. ceranae, the disease signs such as dysen-
tery or crawling behavior or milky white coloration of gut, that are
usually associated with N. apis infection, has never been described
in N. ceranae-infected bees (Fries et al. 2006). The absence of
these disease symptoms in N. ceranae-infected A. mellifera might
reļ¬ect the absence or low intensity of N. ceranae speciļ¬c PCR
signals in the muscles and fat bodies of infected bees, respec-
tively. It is not clear why N. ceranae has different pathological
effects on the host A. mellifera compared with N. apis. Further
studies are warranted to ascertain the pathogenesis of both para-
sites in the A. mellifera.
The sequences of the rRNA operon have been widely used as a
molecular marker for detection of microsporidian infection,
differentiation of closely related species, and estimation of phylo-
genetic relationship among microsporidia. The organization of the
rRNA gene of N. ceranae contains one SSUrRNA gene at the
Fig. 6ā8. Electron-micrographs of longitudinal sections of spores of
Nosema ceranae. 6. Micrograph showing anchoring disk (AD), polaroplast
(P), posterior vacuole (PV), polar ļ¬lament (PF). 7. Micrograph showing
endospore (EN), exospore (EX), plasma membrane (PM), nucleus (N), 20ā
22 isoļ¬lar coils of the PFs. 8. A spore with an extruded polar ļ¬lament
(EPF). Note the more conspicuous PV.
145CHEN ET AL.āCHARACTERIZATION OF NOSEMA CERANAE
5. 50
end, one LSUrRNA gene at 30
end, and an ITS located between
the SSUrRNA and LSUrRNA genes. Parallel comparison of the
rRNA gene sequences of N. ceranae and N. apis showed a se-
quence identity of 92.7% for SSUrRNA, 91.9% for LSUrRNA,
and 48.5% for ITS. Although N. apis and N. ceranae infect the
same host and share similarities in sequences of rRNA gene, our
phylogenetic analysis based on sequences of SSUrRNA showed
that N. apis is not the closest relative of N. ceranae. The same
phylogenetic relationshop of N. apis and N. ceranae was also il-
lustrated in earlier works for characterization of the microsporidian
Fig. 9. Detection of Nosema ceranae by polymerase chain reaction (PCR) ampliļ¬cation of nucleic acids from different tissues. DNA was extracted
from tissues and examined for the presence of N. ceranae-signal by PCR method and electrophoresis. The gel numbers 1ā6 indicate the hypopharyngeal
gland, salivary gland, alimentary canal, Malpighian tubules, muscle, and fat body, respectively; N indicates negative control and P indicates positive
control. The size of PCR fragments is indicated on the right of the gel.
Fig. 10. Phylogenetic tree of microsporidia. Phylogenetic tree of microsporidia infecting insects based on the sequences of the small subunit rRNA
gene and constructed by maximum parsimony analysis under a heuristic search. Trachipleistophora hominis-infecting Homo sapiens was used as an
outgroup. The non-lepidopteran Nosema species are indicated by an asterisk. The reliability of the tree topology is shown by the bootstrap values located
on the tree branches.
146 J. EUKARYOT. MICROBIOL., 56, NO. 2, MARCHāAPRIL 2009
6. species from mosquitoes and spider mites (Becnel et al. 2002;
MuĀØller et al. 2000). Within the same clade, N. ceranae appears to
be more closely related to N. vespula, a parasite infecting wasps,
with 80% bootstrap support. Nosema apis seems to have branched
off earlier and is most closely linked to N. bombi, a parasite in-
fecting bumble bees.
The comparative analysis of rRNA sequences indicated that
ribosomal RNA is conserved and maintains a similar secondary
and tertiary structure for all types of organisms (Gutell, Noller,
and Woese 1986a; Gutell et al. 1986b). While the microsporidian
rRNAs contain some of the characteristic features found in the
vast majority of the eukaryotic rRNAs, the 16S-like and 23S-like
rRNAs of N. ceranae are very unusual. They lack many of the
structural elements present in other nuclear-encoded eukaryotic
rRNAs, and they are signiļ¬cantly shorter in length. For example
the Saccharomyces cerevisiae 16S-like and 23S-like rRNAs are
approximately 1,800 and 3,550 nucleotides in length, the N. cer-
anae 16S-like and 23S-like rRNAs are 1,259 and 2,530 nucleo-
tides in length, respectively. To determine how the reduction in
size of rRNA contributes to the life cycle of the intracellular par-
asite in the host, further studies are needed.
As with many other new and emerging pathogens, we are just
beginning to scratch the surface of understanding how N. ceranae
adopt and establish infection in the new host. Genomic and bio-
chemical characterizations of N. ceranae are currently in progress
to study the roles of parasite genetic variability, host physiological
conditions, and host immune status in the course of infection and
disease.
ACKNOWLEDGMENTS
We would like to thank Michele Hamilton, Bart Smith, and
Andrew Ulsamer for their excellent technical assistance. The
work was supported in part by the 2006 California State Bee-
keepersā Association (CSBA) research fund. R. Gutell was sup-
ported by the National Institutes of Health (GM067317) and the
Welch Foundation (F-1427).
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Received: 12/21/07, 04/02/08, 05/22/08, 07/08/08; accepted: 10/08/08
147CHEN ET AL.āCHARACTERIZATION OF NOSEMA CERANAE