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SummaryMites in the genus Tropilaelaps are native brood parasites of the non-domesticated giant Asian honey bees, Apis dorsata, A. breviligula and A. laboriosa. They spread onto the managed European honey bee (A. mellifera) some time after humans introduced that bee into Asia. Nowadays, A. mellifera is kept for beekeeping throughout Asia and Tropilaelaps mites are one of its most damaging pests. At present, these mites remain confined to Asia and bordering areas but are recognized as emerging threats to world apiculture. In spite of their important pest-status, Tropilaelaps mites remain poorly studied and much remains to be learned about them. The methods reviewed here are intended to assist future research efforts to better understand these parasites. The review begins with an introduction that clarifies why Tropilaelaps mites are worthy of immediate research. It is followed by an outline of the mites' taxonomy and descriptions of various methods (including their pros and cons) by which mites can be coll...

SummaryPollination by honey bees plays a key role in the functioning of ecosystems and optimisation of agricultural yields. Severe honey bee colony losses worldwide have raised concerns about the sustainability of these pollination services. In many cases, bee mortality appears to be the product of many interacting factors, but there is a growing consensus that the ectoparasitic mite Varroa destructor plays the role of the major predisposing liability. We argue that the fight against this mite should be a priority for future honey bee health research. We highlight the lack of efficient control methods currently available against the parasite and discuss the need for new approaches. Gaps in our knowledge of the biology and epidemiology of the mite are identified and a research road map towards sustainable control is drawn. Innovative and challenging approaches are suggested in order to stimulate research efforts and ensure that honey bees will be able to sustainably fulfil their role in the ecosystem.

Adult drone honey bees from 4 Australian breeding lines were reared under similar conditions and examined for semen and sperm production when 14, 21 and 35 days old, during spring, summer and autumn. Almost half (40.5%) of all drones examined did not release any semen when manually everted. For those that released semen, the average volume released per drone was 1.09 μL (range 0.72 (±0.04)-1.12 (±0.04) μL) and the average number of sperms in the semen per drone was 3.63 × 10 6 (range 1.88 (±0.14)- 4.11 (±0.17) × 10 6 ). The release of semen was dependent on breeding line and age (P < 0.05), but not on the rearing season. The volume of semen released per drone was dependent on season, age, and breeding line (P < 0.05), while the concentration of sperm in the semen was dependent on season and breeding line (P < 0.05). Hence our data indicate that genetics underpins the maturation of drone honey bees as well as the volume of semen they release and the concentration of sperm in that semen. Apis mellifera / drones / semen production / sperm

Two new taxa have been added to the existing molecular phylogenies of the genus Apis. The new phylogeny supports the accepted phylogenetic relationships of {dwarf honey bees [giant honey bees (cavity-nesting honey bees)]}. Based on Bayesian and maximum parsimony trees, our analysis supports recognition of Apis indica, the Plains Honey Bee of south India, as a separate species from A. cerana. Our analysis also supports recognition of the Giant Philippines Honey Bee, A. breviligula, as a separate species from A. dorsata.

The invasion of the Western honey bee, Apis mellifera, by Varroa destructor is attributed to two mitochondrial haplotypes (K and J) that shifted last century from their primary host the Eastern honey bee, A. cerana, in north-east Asia. Here, mitochondrial DNA sequences (cox1, cox3, atp6 and cytb: 2700 base pairs) were obtained from mites infesting both Eastern and Western honeybees (respectively 21 and 11 colonies) from Asia including regions where the shifts first occurred. A total of eighteen haplotypes were uncovered in Asia (11 on A. cerana and 7 on A. mellifera). Two new variants of the K haplotype and two of the J haplotype were found on Western honeybees in what appeared to be well-established infestations. New haplotypes may represent a potential threat to A. mellifera worldwide. The extreme lack of polymorphism in the K and J haplotypes outside of Asia, can now be plausibly explained as being due to genetic 'bottlenecks' that occurred in Asia before and after mites shifted from their original Eastern honeybee host. Apis mellifera / Apis cerana / Varroa / mitochondrial DNA/diversity

Varroa jacobsoni was first described as a natural ectoparasitic mite of the Eastern honeybee (Apis cerana) throughout Asia. It later switched host to the Western honeybee (A. mellifera) and has now become a serious pest of that bee worldwide. The studies reported here on genotypic, phenotypic and reproductive variation among V. jacobsoni infesting A. cerana throughout Asia demonstrate that V. jacobsoni is a complex of at least two different species. In a new classification V. jacobsoni is here redefined as encompassing nine haplotypes (mites with distinct mtDNA CO-I gene sequences) that infest A. cerana in the Malaysia–Indonesia region. Included is a Java haplotype, specimens of which were used to first describe V. jacobsoni at the beginning of this century. A new name, V. destructor n. sp., is given to six haplotypes that infest A. cerana on mainland Asia. Adult females of V. destructor are significantly larger and less spherical in shape than females of V. jacobsoni and they are also reproductively isolated from females of V. jacobsoni. The taxonomic positions of a further three unique haplotypes that infest A. cerana in the Philippines is uncertain and requires further study. Other studies reported here also show that only two of the 18 different haplotypes concealed within the complex of mites infesting A. cerana have become pests of A. mellifera worldwide. Both belong to V. destructor, and they are not V. jacobsoni. The most common is a Korea haplotype, so-called because it was also found parasitizing A. cerana in South Korea. It was identified on A. mellifera in Europe, the Middle East, Africa, Asia, and the Americas. Less common is a Japan/Thailand haplotype, so-called because it was also found parasitizing A. cerana in Japan and Thailand. It was identified on A. mellifera in Japan, Thailand and the Americas. Our results imply that the findings of past research on V. jacobsoni are applicable mostly to V. destructor. Our results will also influence quarantine protocols for bee mites, and may present new strategies for mite control.

SUMMARYThis paper demonstrates the presence of two genetically distinct populations of Varroa jacobsoni that show contrasting abilities to reproduce on Apis mellifera and can be identified easily using simple molecular techniques. When Australian-reared A. mellifera sister queens were transported to Germany and Papua New Guinea and established in varroa-infested colonies, the female V. jacobsoni in Germany readily reproduced on their brood whereas in Papua New Guinea they did not. These observations indicate that the cause of the reported inability of female V. jacobsoni to reproduce on A. mellifera in Papua New Guinea does not rest with the resident A mellifera population. DNA sequences of a 454 base-pair region of the cytochrome oxidase subunit 1 (CO1) gene obtained from reproducing V. jacobsoni collected from A. mellifera colonies in Germany differed by 6.8% from similar sequences obtained from non-reproducing V. jacobsoni from A. mellifera colonies in Papua New Guinea. The presence of a Xho I restrict...

SUMMARYA sensitive culture technique for detecting viable spores and mycelial elements of the chalkbrood fungus, Ascosphaera apis, is described. The technique involves embedding honey or distilled water containing A. apis spores or mycelial elements in 15 ml of sterile liquid nutrient agar medium (10 g yeast, 10 g glucose, 13.5 g KH2PO4, 10 g soluble starch and 20 g agar) cooled to 60°C. This medium is then poured on a 7-ml layer of similar but solid agar medium in a standard 8.5-cm petri dish and allowed to solidify by cooling. The medium is incubated in an anaerobic environment at 37°C for 24 h, and then incubated in an aerobic environment for up to 9 days at 37°C. It is examined daily for A. apis growth. The technique facilitated the detection of viable A. apis in honey and was used to show that many pre-packaged retail honeys contain viable A. apis. The technique was also used to show that honey may be rendered free of viable A. apis by holding it in water baths at 65°C for 8 hours or at 70°C for 2 hours.

SummaryVery rapidly after Varroa destructor invaded apiaries of Apis mellifera, the devastating effect of this mite prompted an active research effort to understand and control this parasite. Over a few decades, varroa has spread to most countries exploiting A. mellifera. As a consequence, a large number of teams have worked with this organism, developing a diversity of research methods. Often different approaches have been followed to achieve the same goal. The diversity of methods made the results difficult to compare, thus hindering our understanding of this parasite. In this paper, we provide easy to use protocols for the collection, identification, diagnosis, rearing, breeding, marking and measurement of infestation rates and fertility of V. destructor. We also describe experimental protocols to study orientation and feeding of the mite, to infest colonies or cells and measure the mite's susceptibility to acaricides. Where relevant, we describe which mite should be used for bioassays since their beha...

La presence et la reproduction de l'acarien parasite V jacobsoni ont ete etudiees dans des colonies d'Apis cerana Fabr et d'A mellifera en Papouasie Nouvelle-Guinee (PNG) a differentes reprises entre 1991 et 1993, a Irian Jaya en septembre 1991 et a Java en octobre 1991. Dans les colonies d'A cerana, comme dans les colonies d'A mellifera, il y avait significativement plus de couvain de mâles que de couvain d'ouvrieres infeste par des acariens adultes, quelle que soit la localite. Dans les colonies des 2 especes, des acariens adultes vivants ont ete observes sur tous les stades de developpement du couvain de mâles et d'ouvrieres

A study to determine the effect of pathogens on pollen collection by colonies of the honey bee, Apis mellifera L., is reported. Colonies fed preparations of 50% sucrose containing sacbrood virus particles or Nosema apis Zander spores collected significantly less pollen than colonies fed only the sucrose solution. These findings have important implications for the preparation and use of honey bee colonies for pollinating agricultural crops.

SummaryThe occurrence and distribution of pests and pathogens of the honeybee, Apis mellifera L., in Fiji, surveyed during -10—November 1986, are reported. American foulbrood disease was detected in 33 (47%) hived colonies at one locality but in only 1 (0·8%) of all other colonies examined in other localities. Infestations of wax moth (Galleria mellonella or Achroia grisella) were observed in 139 (70%) hived colonies. The cane toad Bufo marinus was also observed as a pest of bee colonies.Chronic bee paralysis virus (CBPV) and sacbrood virus (SBV) were present in 30 (31%) and 76 (79%) hived colonies, respectively. Bee virus X, bee virus Y, black queen cell virus (BQCV), Kashmir bee virus (KBV), and slow bee paralysis virus were also detected in these colonies but they were less common than CBPV and SBV. Inapparent infections of SBV, BQCV and KBV were detected in seemingly healthy bees from hived colonies. SBV and CBPV were also detected in feral colonies. The Fijian KBV isolates were serologically distingu...

Mites in the genus Tropilaelaps are parasites of social honeybees. Two species, Tropilaelaps clareae and T. koenigerum, have been recorded and their primary hosts are presumed to be the giant honeybees of Asia, Apis dorsata and A. laboriosa. The most common species, T. clareae, is also an economically important pest of the introduced Western honeybee (A. mellifera) throughout Asia and is considered an emerging threat to world apiculture. In the studies reported here, genetic (mtDNA CO-I and nuclear ITS1-5.8S-ITS2 gene sequence) and morphological variation and host associations were examined among Tropilaelaps isolates collected from A. dorsata, A. laboriosa and A. mellifera throughout Asia and neighbouring regions. The results clearly indicate that the genus contains at least four species. Tropilaelaps clareae, previously assumed to be ubiquitous in Asia, was found to be two species, and it is here redefined as encompassing haplotypes (mites with distinct mtDNA gene sequences) that parasitise native A. dorsata breviligula and introduced A. mellifera in the Philippines and also native A. d. binghami on Sulawesi Island in Indonesia. Tropilaelaps mercedesae n. sp., which until now has been mistaken for T. clareae, encompasses haplotypes that, together with haplotypes of T. koenigerum, parasitise native A. d. dorsata in mainland Asia and Indonesia (except Sulawesi Island). It also parasitises introduced A. mellifera in these and surrounding regions and, with another new species, T. thaii n. sp., also parasitises A. laboriosa in mountainous Himalayan regions. Methods are described for identifying each species. These studies help to clarify the emerging threat of Tropilaelaps to world apiculture and will necessitate a revision of quarantine protocols for countries that import and export honeybees.

International audience; A total of 24 Varroa mite samples were collected from A. mellifera and A. cerana colonies in 17 Provinces in China and subjected to DNA or restriction enzyme analyses. All Varroa mites from A. mellifera were identified as the Korean genotype of Varroa destructor Anderson and Trueman, confirming earlier finding that V. jacobsoni is not present in China, and a single genotype of Varroa is infecting the A. mellifera bees. Mites collected from A. cerana colonies at 2 locations in Guangdong Province were the China genotype of V. destructor. Mites collected from A. cerana colonies in 3 apiaries at Dayao County, Yunnan Province, and from 5 colonies at Mongzi County, Yunnan Province, were a new genotype, which was named the China 2 genotype of V. destructor. Mites collected from A. cerana colonies in 4 apiaries at Xishuangbanna, Yunnan Province, were the Vietnam genotype of V. destructor. These results should help future mite control and bee quarantine efforts in China.

International audience; In the debris of five Apis dorsata colonies at a single site in Sabah, Borneo we found the mites Tropilaelaps clareae, Tropilaelaps koenigerum, Varroa rindereri, Varroa jacobsoni and Euvarroa wongsirii. Most were T. clareae, but T. koenigerum were also quite common. The V. rindereri specimens belonged to the same haplotype as described previously from A. koschevnikovi from Borneo. However, the V. jacobsoni belonged to a new haplotype, which we named the 'Borneo 2 haplotype of V. jacobsoni'. Of the mites detected in the debris, 84% of the T. clareae and 57% of the T. koenigerum were damaged. Inspection of 1673 brood cells of two A. dorsata colonies at the same site resulted in adult T. clareae and T. koenigerum together with their offspring (nymphs). The percentage of infested drone and worker cells did not differ, nor did the number of mites per cell: 6.0 $\pm$ 1.6 in worker brood and 6.1 $\pm$ 1.9 in drone brood (n = 10). We found no Varroa mites in the inspected brood cells, suggesting that the mites do not reproduce in A. dorsata and indicating that interspecific mite transfer occurs between sympatric Apis species in Borneo.

International audience; Note scientifique sur la différence génétique des acariens Varroa présents sur Apis mellifera et Apis cerana.

En complement a des etudes anterieures, la presence et la reproduction de l'acarien V jacobsoni dans des cellules de couvain opercule (prenymphes et stades ulterieurs) ont ete etudiees sur des colonies d'A cerana et d'Apis mellifera en Papouasie Nouvelle-Guinee (PNG), a Irian Jaya, i) pour des colonies d'A cerana indigenes et ii) pour des colonies d'apis mellifera importees de Java debut 1995. Dans toutes les localites il y avait proportionnellement plus de cellules de mâles que de cellules d'ouvrieres infestees par des femelles adultes de varroa, que ce soit dans les colonies d'A cerana ou d'Apis mellifera (tableaux I et II). Dans toutes les colonies d'A cerana, independamment de la localite, les femelles de varroas ne se sont raproduites dans les cellules de mâles (tableau I). Pour les colonies d'A melifera, en revanche, le resultat a varie selon la localite et l'annee. En PNG et a Irian Jaya, les femelles de varroas ne se reproduises ni dans les cellules de mâles, ni dans celles d'ouvrieres. A Java il en a ete de meme en 1992, ainsi que dans une etude anterieure faite en 1991. Mais en 1993, les femelles de varroas se sont reproduites dans les cellules d'ouvrieres et les annees suivantes, a la fois dans les cellules d'ouvrieres et celles des mâles (tableau II). A Biak et Yapen egalement, en 1995, elles se sont reproduites dans les deux types de cellules. On propose deux hypotheses comme cause possible de ce changement. Selon la premiere, les conqtraintes qui empechaient les femelles de V jacobsoni de se reproduire dans les colonies d'A mellifera a Java avant 1992 ont disparu entre janvier 1992 et fevrier 1993, leur permettant ainsi de commencer a se reproduire. Selon l'autre hypothese, une nouvelle souche de V jacobsoni, capable de se reproduire dans les cellules de couvain opercule, a ete introduite a Java entre janvier 1992 et fevrier 1993.