Detection of Paenibacillus larvae subspecies larvae spores in naturally infected bee larvae and artificially contaminated honey by PCR

American foulbrood (AFB), a severe bacterial disease of honeybee brood, has recently been found in Uruguayan apiaries. Detection of the causative agent, Paenibacillus larvae subspecies larvae, is a very important concern in order to prevent disease dissemination and decrease of honey production. Since spores are the infective forms of this pathogen, in the present work we report the use of polymerase chain reaction (PCR) to detect P. l. subsp. larvae spores from in vitro cultures, larvae with clinical symptoms and experimentally contaminated honey. The set of primers was designed based on the published P. l. subsp. larvae 16S rRNA gene. Using this approach we could amplify the pathogen DNA and obtain a great sensitivity and a notable specificity. Detection limit for spore suspension was a 10^–2 dilution of template DNA obtained from 32 spores, as determined by plate count. For artificially contaminated honey, we could detect the PCR product at a 10^–3 dilution of template DNA obtained from 170 spores. In addition, when PCR conditions were set to improve specificity, we were able to amplify P. l. subsp. larvae DNA selectively and no cross-reactions were observed with a variety of related bacterial species, including P. l. subsp. pulvifaciens. Since spore detection is very important to confirm the presence of the disease, this method provides a reliable diagnosis of AFB from infected larvae and contaminated honey in a few hours.     

Suppression of phototaxis in silkworm larvae

A study of the olfactory and visual organs of the larvae of the silkworm (Bombyx mori), using electrophysiological and surgical techniques, indicates that olfactory stimuli from mulberry leaves, conducted through the antennae or the maxillary palps, cause continuous suppression of the phototactic response, and that the central nervous system plays an important role in this ‘control’ of phototaxis. Such phototactic suppression lasts for 30 h in fifth instar larvae, even after mulberry leaves have been removed.     

Jet-propulsion in anisopteran dragonfly larvae

Summary Jet-propulsion in dragonfly larvae is achieved by the rapid ejection of water from a specialised rectal chamber via the anus, at a frequency of up to 2.2 cycles/s. Movement, forward thrust and muscular activity have been recorded in restrained and free-swimming larvae. Forward thrusts of up to 1.5 g wt result from the expiratory phases of cycles lasting 0.1 to 0.4 s. Swimming velocities are in the order of 10 cm/s. The following muscles are shown to be active during expiratory phases of jetting: anterior, posterior and respiratory dorso-ventrals; primary and secondary longitudinal tergals; lateral primary longitudinal sternopleural; dorso-ventral oblique; ventral adductors of the anal appendages. The sub-intestinal muscle is active during the inspiratory phases of jetting. Activity recorded is compared with that found during normal ventilation. The larval jet-propulsive mechanism is compared with that of certain cephalopods and found to be very effective for the larva's relatively small size.R. S. P. wishes to thank the Science Research Council for a studentship during the tenure of which the above work was carried out.     

Photoresponse in allis shad larvae

Allis shad Alosa alosa photoresponse was investigated in 3 day-old to 1 month-old larvae. Regardless of age, larvae strongly preferred the lit compartment in aquaria experiments. Such response was consistent with other experimental studies which have suggested a pelagic behaviour of allis shad larvae from the time of hatch.     

Feeding mechanisms in anuran larvae

Larvae of the neotropical frogs Phyllomedusa are distinctive in that they feed normally in mid-water on phytoplankton, maintaining neutral buoyancy by means of an independently beating tail filament. The feeding mechanism of larval Phyllomedusa trinitatis was studied morphologically and experimentally. The primary feeding mechanism involves a buccal rasp which may in some circumstances render food into small particles, a pumping mechanism which forces water through the buccal cavity and the gill filters, an entanglement system which traps particles in mucous strings produced in special organs, and the formation of mucous cords which transport particles to the oesophagus. In mid-water feeding and surface feeding the buccal rasp serves only its other function in preventing backflow of the respiratory stream. The primary feeding mechanism is discussed and compared with schemes proposed for Rana temporaria and R. agilis. Little agreement exists between these schemes and that which is here proposed. It is concluded that the primary feeding mechanism is the same in the three forms but that there are behavioural differences in feeding generally. Some comment is made on the primary feeding mechanism in the Microhylidae.     

Paenibacillus larvae larvae spores in honey samples from Uruguay: a nationwide survey

American foulbrood is a severe bacterial disease affecting larvae of the honeybee Apis mellifera and it is caused by Paenibacillus larvae larvae. The disease is present worldwide and cases have been reported in almost all the beekeeping regions of the five continents. During 2001 and 2002 we carried out a nationwide study to assess the presence and amount of P. l. larvae spores in honey samples from Uruguay, combining classic bacteriological, and molecular approaches. The distribution of P. l. larvae spores in honey of the whole country showed a clear pattern and may provide useful data for a control and prevention strategy of American foulbrood.     

Sexing schistosome larvae

Biological and immunological factors may influence changes in sex ratios at different points of the schistosome life cycle, resulting in the fact that female schistosomes are significantly outnumbered by males in chronic infections of snails and mammalian hosts. Analysis of this phenomenon has long been hampered by shortcomings in the methods used to determine sex ratios. Here, Robin Gasser describes recently developed molecular methods for sexing schistosome larvae. These have opened the way towards understanding why sex ratios become male biased and allow a proper assessment of its consequences in the epidemiology, diagnosis and pathology of infection.