Black queen cell virus and Nosema ceranae coinfection in Africanized honey bees from southern Brazil / Coinfecção por vírus da realeira negra (BQCV) e Nosema ceranae em abelhas africanizadas no sul do Brasil

Bees are fundamental in several aspects, especially in relation to plant biodiversity and pollination. Recently, immense losses are being faced in the number of Brazilian colonies, mainly in southern states of the country, which has a strong beekeeping activity. There are indications that, among the reasons for the losses, pathogens that affect the health of bees may be involved. Among them, the microsporidium Nosema and the black queen cell virus (BQCV) stand out for their prevalence. In this study, 92 colonies of 17 apiaries from southern Brazil were evaluated for infection by Nosema ceranae, Nosema apis and BQCV. Nucleic acid extractions and cDNA synthesis were performed from adult bee samples, followed by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and multiplex PCR. Eight BQCV positive samples were subjected to sequencing. The results showed that N. ceranae and BQCV are circulating in the Southern region of the country, which may be the reason for the loss of colonies. N. apis was not found. N. ceranae was found in 57.6% (53/92) of the colonies and BQCV in 32.6% (30/92). Co-infection was found in 25% (23/92) of the colonies studied, a factor that is suggested to be reducing the hosts' longevity due to the synergistic action of the pathogens. The samples submitted to sequencing indicated similarity of 96.8 to 100% between them, in addition to strong similarity with sequences from Asia, United States, Germany and Peru. This study reports the circulation of N. ceranae and BQCV in apiaries in southern Brazil, in addition to being the first phylogenetic analysis of the Brazilian BQCV sequence.(AU)

As abelhas mostram-se fundamentais em diversos aspectos, especialmente com relação à biodiversidade de plantas e polinização. Recentemente, estão sendo enfrentadas imensas perdas no número de colônias brasileiras, principalmente nos estados do sul do país, com forte atividade apícola. Há indicativos de que, dentre as razões para as perdas, possam estar envolvidos patógenos que afetam a saúde das abelhas. Dentre eles, o microsporídio Nosema e o vírus da realeira negra (BQCV) destacam-se pela prevalência. Neste estudo, foram avaliadas 92 colônias, de 17 apiários do sul do Brasil, a respeito da infecção por Nosema ceranae, Nosema apis e BQCV. Foram realizadas extrações de ácidos nucleicos e síntese de cDNA a partir de amostras de abelhas adultas, seguidos de Reação em Cadeia da Polimerase-Transcriptase Reversa (RT-PCR). Oito amostras positivas para BQCV foram submetidas a sequenciamento. Os resultados mostraram que N. ceranae e BQCV estão circulando na região sul do país, podendo ser a razão para as perdas de colônias. N. apis não foi encontrado. N. ceranae foi encontrado em 57.6% (53/92) das colônias e BQCV em 32.6% (30/92). Foi encontrada coinfecção por ambos em 25% (23/92) das colônias estudadas, fator que sugere a diminuição da longevidade do hospedeiro por ação sinérgica dos patógenos. As amostras submetidas ao sequenciamento indicaram similaridade de 96.8 a 100% entre elas, além de forte similaridade com sequências da Ásia, Estados Unidos, Alemanha e Peru. Este estudo relata a circulação de N. ceranae e BQCV nos apiários do sul do Brasil, além de ser a primeira análise filogenética da sequência do BQCV brasileiro.(AU)

Efficient Method for the Rapid Purification of Nosema ceranae Spores

Nosema ceranae is an obligate intracellular fungal parasite that causes mortality in honey bees and enhances the susceptibility of honey bees to other pathogens. Efficient purification of Nosema spores from the midgut of infected honey bees is very important because Nosema is non-culturable and only seasonably available. To achieve a higher yield of spores from honey bees, in this study, we considered that the initial release of spores from the midgut tissues was the most critical step. The use of 2 mm beads along with enzymatic treatment with collagenase and trypsin enhanced the homogenization of tissues and the yield of released spores by approximately 2.95 times compared with the use of common 3 mm beads alone. The optimal time for the enzyme treatment was determined to be 1 hr as measured by the yield and viability of the spores. A one-step filtration using a filter paper with an 8–11 µm pore size was sufficient for removing cell debris. This method may be useful to purify not only N. ceranae spores but also other Nosema spp. spores.

Prevalence of Nosema and Virus in Honey Bee (Apis mellifera L.) Colonies on Flowering Period of Acacia in Korea

Honey production from approximately 1.6 million colonies owned by about 199,000 Korean beekeepers was almost 23,000 metric tons in 2009. Nosema causes significant losses in honey production and the virus decreases population size. We initiated a survey of honey bee colonies on the blooming period of Acacia to determine the prevalence of Nosema and virus in 2011. Most Korean beekeepers have moved from the south to north of Korea to get Acacia nectar for 2 mon. This provided a valuable opportunity to sample bees originating from diverse areas in one location. Twenty hives owned by 18 beekeepers were sampled in this year. Nosema spore counts ranged from zero to 1,710,000 spores per bee. The average number of nosema spores per bee was 580,000. Approximately 95% of the colonies were infected with Nosema, based on the presence of spores in the flowering period of Acacia. This indicates that Nosema is the predominant species affecting honeybee colonies. Also, the seven most important honeybee viruses were investigated by reverse transcription-PCR. Among them, four different viruses were detected in samples. Black queen cell virus was present in all samples. Chronic bee paralysis virus was detected in 10% of samples. Deformed wing virus was present in only 5% of the samples. Prevalence of Sacbrood virus was 15%. However, Cloudy wing virus, Israel acute paralysis virus and kashmir bee virus were not detected in any of samples.

Nosema like organisms found in an immunocompetent human patient: first report in Central América

Stool samples of a immunocompetent hypertense 61 year old woman were stained by the Weber et al. method and studied by electronic microscopy. Mature microsporidia spores were found and according to their diplokaryotic muclei, as well as the typical polar filament with 12 coils, the organisms was classified as Nosema like genus

Microspiridiosis: una parasitosis de reciente adaptación al hombre

Se hace un análisis general de la literatura y del trabajo de los autores sobre microsporidiosis, una parasitosis que se ha puesto en boga a raíz de los problemas de inmunosupresión, especialmente en personas infectadas con el virus del SIDA. Se hace una revisión histórica, se describe básicamente la morfología de los géneros más importantes de los microsporidios y se comentan algunos aspectos de la epidemiología. Se revisan los cambios patológicos debidos a la infección con estos parásitos, los métodos de diagnóstico más usados, el tratamiento y la prevención contra esta parasitosis. Se enfatiza sobre el hallazgo por primera vez en Costa Rica y probablemente en Centro América, de casos humanos de infección con Enterocytozoon bieneusi y con el género Nosema. Palabras claves: Microsporidia, microsporidiosis, Enterocytozoon, Encephalitozoon, Pleisthphora, Nosema

Impacto de nuevas metodologías en el estudio de parásitos: V. Dinámica del disparo del filamento polar en el microsporidio Nosema algerae, analizado por videomicroscopía de contraste acentuado electrónicamente / Impact of new methodologies in the study of parasites: V. Dynamics of the polar filament shot in the microsporidium Nosema algerea, analyzed by electronically enhanced contrast videomicroscopy

Por medio de video-microscopía de contraste acentuado electrónicamente, se consiguió el primer análisis cinemático de la descarga del filamento polar y el esporoplasma por esporas de un microsporidio. La estimulación in vitro de esporas de Nosema algerea, un parásito de los mosquitos, provoca la salida explosiva del filamento polar con una velocidad instantánea máxima de 105 µm/s en promedio, seguida por la expulsión del esporoplasma en el extremo del filamento luego de un lapso variable con un máximo de 500 ms. La descarga total se completa en menores de 2 s. La morfología de la parte del filamento ya descargada en cada instante no cambia durante la salida, lo que sugiere que el alargamiento ocurre tan solo en el extremo distal, conforma a la opinión de que el filamento es extruido por eversión. Por lo común, la longitud del filamento disminuye entre 5 y un 10 por ciento después de la expulsión del esporoplasma, lo que indica elasticidad del material constitutivo y presurización interna durante el proceso. Una vez liberado el esporoplasma aumenta de volumen, como es de esperar de una alta presión osmótica residual que, de acuerdo con la hipótesis prevaleciente, es ocasionada por la estimulación. Los resultados apoyan el modelo de que las esporas de los microsporidios germinan cuando el estímulo causa un aumento de presión osmótica interna, que a su vez determina un influjo de agua de manera que la presión hidrostática se eleva y finalmente rompe la tapa polar de la espora, por donde son entonces expulsados el filamento y enseguida el esporoplasma.

Actualizaciones sobre microsporidiosis en el hombre / Update on microsporidiosis in man

The importance of microsporidium as an opportunistic agent in immunocompromised and AIDS patients is reviewed. Five strains of the agent have been described: Encephalitozoon, Enterocytozoon, Nosema, Pleistophora and Septata. The clinical presentation may be as 1) Generalized infections with multisystemic involvement, specially of the central nervous system; 2) Intestinal, that is the most important and frequent localization in man, and that may cause death in AIDS patients; 3) Ocular, that affects cornea, conjunctiva and may extend to paranasal sinuses; 4) Liver and biliary tract infection with granulomatous lesions, hepatic necrosis or sclerosing colangitis and 5) Muscular, affecting skeletal muscle. The diagnosis is difficult and is established finding spores in the affected tissues with light or electron microscopy. Lately, the diagnosis of intestinal microsporidiosis is made looking for faecal spores. The resistant wall of spores hampers treatment. However, good results are obtained with albendazole in intestinal microsporidiosis