Steinernema brazilense n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from Mato Grosso, Brazil

A new entomopathogenic nematode, Steinernema brazilense n. sp., was isolated from a single soil sample collected from a natural forest in Mato Grosso do Sul state, Brazil. S. brazilense n. sp. is characterized morphologically by features of infective juveniles (IJ), males and females. For the IJ, body length averaging 1157 (1023-1284) μm, distance from anterior end to excretory pore 95 (87-102) μm, from anterior end to end of esophagus 148 (139-153) μm, tail length 85 (80-104) μm, D% and E% values 63 (58-70) and 106 (95-118.0), respectively. Lateral field pattern variable; the formula for the arrangement of ridges from head to tail is: 2, 4, 6, 8, 6, 2. For the male, the diagnostic characters include spicule averaging 83 (75-89) μm; D% about 65; the ratio SW% about 192. The length of spicule head is greater than width. Lateral field with one narrow ridge. First generation females are characterized by the presence of a ventral postanal swelling. S. brazilense n. sp. is morphologically close to Steinernema diaprepesi. It can be differentiated from S.diaprepesi by its longer IJ body length (1157 vs 1002 μm), longer distance from anterior end to excretory pore (110 vs 75 μm), a longer tail length (103 vs 83 μm); males of the new species with longer spicule (83 vs 79 μm). The new species can be distinguished further from other members of Steinernema glaseri group by characteristics of rDNA of ITS and D2D3 regions.     

Studies on Procamallanus (Spirocamallanus) Pereirai annereaux, 1946 (Nematoda: Camallanidae), with new host records and new morphological data on the larval stages.

Larval stages and adults of Procamallanus (Spirocamallanus) pereirai Annereaux, 1946 are described from naturally infected Paralonchurus brasiliensis (Steindachner) (Sciaenidae) from the coast of the State of Rio de Janeiro, Brazil. The translucent first-stage larvae have a denticulate process at the anterior end, no buccal capsule or esophagus undifferentiated into anterior muscular and posterior glandular parts and an elongate tail; third-stage larvae have a tail with three terminal projections, a buccal capsule divided into an anterior portion with 12-20 ridges running to the left and a posterior smooth portion, and an esophagus with muscular and glandular regions. Fourth-stage larvae exhibit a buccal capsule lacking a distinct basal ring with ridges running to the right and a tail with two terminal processes, as in adults. New host records are reported and their role in its life-cycle are discussed.     

Bemerkungen zur Biologie sowie Haltung und Vermehrung des Sokotra-Riesengeckos, Haemodracon riebeckii (Peters, 1882)

The Socotra Giant Gecko, Haemodracon riebeckii, is the largest species of lizard on Socotra Island. The nocturnal, arboreal and rupiculous living geckos are omnivorous. Two pairs were kept in terrariums and were fed with various insects (crickets, locusts, cockroaches), sweet fruits and other feeding stuff (such as meat, fish). Temporarily H. riebeckii was kept together with other lizards (Eublepharis macularius, Trachylepis socotrana), without any signs of aggressive behaviour. Juveniles and adults of both sexes are able to produce a sound. These acoustic signals seem to be related to predators, because never any intraspecific function could be observed. Within seven years of captive breeding two females produced 253 eggs. Usually two white and sticky soft-shelled eggs were laid within one clutch, more rarely a single egg was laid. The two eggs of a clutch were always laid on the same day. H. riebeckii belongs to the geckos that bury their eggs and practice some brood care, but no special parental care. The female is able to proof with her hind legs the deep and shape of a hollow in the substrate to bury the eggs, which were buried in a sticky and soft-shelled condition. They are oval in shape (egg length 16.4-19.8 mm, egg width 12.4-17.8 mm, quotient EL:EW 1.22±0.05) and have in the beginning a weight ranging from 1.7100 to 2.5201 g. As typical for geckos with hard-shelled eggs the egg weight decreases during the incubation period. The loss can be between 5.59 to 30.29%. The development of eggs up to hatching of young depends upon temperature and the germinal stage in the laid egg. The time difference between the hatching of the young within one clutch of two eggs was usually 1 to 5 days. In some cases there were, however, longer differences (up to 61 days), which are probably caused by different developmental stages of the embryos during the time of egg laying. The shortest incubation period recorded during our investigations was 83 days for eggs incubated at constant temperature of 28 to 29.5 °C and the longest 359 days at 26 to 26.5 °C. Constant high incubation temperatures caused a premature hatching of young. In normal hatched young were the yolk sac retracted and the navel closed. In premature hatched young were the yolk not resorbed and the mortality within the first three month comparatively high. The snout-vent length (SVL) of newly hatched young is from 27 to 39 mm and the tail length (TL) from 25 to 38 mm (SVL:TL index 0.90-1.27), the weight is from 0.7688 to 1.5366 g. Young specimens are distinguished from adults by the brown/white striped lower jaw and the white-banded tail. Young which hatched in the terrarium were eaten by the adults. A loss of young can be avoided if they are raised individually.     

Viral pyrogen affects thermoregulatory behavior of wintering Planorbarius corneus (L.) snails (Mollusca: Gastropoda)

Exogenous pyrogens, isolated from viruses, bacteria and fungi, are widely used in research on fever in endotherms. The present study investigates the use of viral pyrogen poly (I:C) for inducing the symptoms of so-called behavioral fever in ectothermic animals Planorbarius corneus (Mollusca: Gastropoda). The observations of thermoregulatory behavior of the snails placed in the temperature gradient showed that the snails which were injected with 750 ng g⁻¹ polyinosinic-polycytidylic acid-poly (I:C), after remaining latent for more than 24 h, moved to warmer areas (with the temperature around 30 °C) and stayed there until the end of the experiment. Transferred to the culture, they lived for a period similar to that of the control snails. Our observations indicate that P. corneus snails can exhibit a symptoms of fever response to poly (I:C) and could be used as model animals in the research on behavioral fever in invertebrates.     

Diapause induction, maintenance and termination in the rice stem borer Chilo suppressalis (Walker)

The rice stem borer, Chilo suppressalis, enters facultative diapause as fully grown larvae in response to short-day conditions during the autumn. Our results showed that the critical night length for diapause induction in C. suppressalis was between 10 h 22 min and 10 h 45 min at 22, 25 and 28 °C, 11 h 18 min at 31 °C, and between 10 h 5 min and 10 h 20 min under field conditions (average temperature ranged from 27.2 to 30.7 °C). The diapause incidence declined in ultra-long nights (18-22 h scotophases) and DD, and increased in ultra-short nights (2-6 h scotophases) and LL. Moreover, we found that the third instar was the stage most sensitive to the photoperiod, and night length played an essential role in the initiation of diapause. Night-interruption experiments with a 1-h light pulse at LD 12:12 (light 12:dark 12) exhibited two troughs of diapause inhibition, with one occurring in early scotophase and the other in late scotophase. Field observations for six years showed that most larvae entered winter diapause in August in response to declining day lengths, despite the high temperatures prevailing during August. By periodically transferring the field-collected overwintering larvae to different photoperiods and temperatures, the results showed that photoperiod had a significant influence on diapause development during the early phase of diapause, while high temperature significantly accelerated the termination of larval diapause.     

Effect of temperature,photoperiod, and feeding on the rate of tail regeneration in a semiaquatic plethodontid salamander

Salamander tail autotomy improves survival, but loss of the tail can subsequently be costly. For example, burst swimming speed is significantly slower after autotomy in desmognathan salamanders, which may increase predation risk in aquatic habitats. However, any long-term cost of tail loss is contingent on the rate of tail regeneration. To examine variation among seasons and environments in the cost of tail autotomy, we tested the effect of temperature, photoperiod, and feeding on tail-length re-growth in the semiaquatic plethodontid salamander Desmognathus conanti. Eight experimental groups (n=15 each, equivalent in body size) were tested. After acclimation for four weeks at one of two temperatures (either 10 °C or 20 °C) and one of two photoperiods (either L:D 9.5:14.5 h or 14.5:9.5 h), 60% of the tail length was autotomized for each individual. After autotomy, each experimental group was maintained under unique conditions of temperature (either 10 °C or 20 °C), photoperiod (either L:D 9.5:14.5 h or 14.5:9.5 h), and feeding (either fasting or weekly feeding). The length of the regenerated tail portion for each individual was measured each week until the group with the fastest re-growth had regenerated 50% of the lost tail length. Low temperature had a large, negative effect, fasting had a small, negative effect, but photoperiod had no significant effect on tail re-growth. The large thermal effect resulted from a combination of delayed initiation of tail-length re-growth and reduced regeneration rate thereafter at low temperature. We conclude that the cost of salamander tail autotomy differs among seasons and environments based on variation in temperature and food availability.     

Effects of osmolality on sperm morphology, motility and flagellar wave parameters in Northern pike (Esox lucius L.)

Northern pike (Esox lucius L.) spermatozoa are uniflagellated cells differentiated into a head without acrosome, a midpiece and a flagellar tail region flanked by a fin structure. Total, flagellar, head and midpiece lengths of spermatozoa were measured and show mean values of 34.5, 32.0, 1.32, 1.17 μm, respectively, with anterior and posterior widths of the midpiece measuring 0.8 and 0.6 μm, respectively. The osmolality of seminal plasma ranged from 228 to 350 mOsmol kg⁻¹ (average: 283.88 ± 33.05). After triggering of sperm motility in very low osmolality medium (distilled water), blebs appeared along the flagellum. At later periods in the motility phase, the tip of the flagellum became curled into a loop shape which resulted in a shortening of the flagellum and a restriction of wave development to the proximal part (close to head). Spermatozoa velocity and percentage of motile spermatozoa decreased rapidly as a function of time postactivation and depended on the osmolality of activation media (P < 0.05). In general, the greatest percentage of motile spermatozoa and highest spermatozoa velocity were observed between 125 and 235 mOsmol kg⁻¹. Osmolality above 375 mOsmol kg⁻¹ inhibited the motility of spermatozoa. After triggering of sperm motility in activation media, beating waves propagated along the full length of flagella, while waves appeared dampened during later periods in the motility phase, and were absent at the end of the motility phase. By increasing osmolality, the velocity of spermatozoa reached the highest value while wave length, amplitude, number of waves and curvatures also were at their highest values. This study showed that sperm morphology can be used for fish classification. Sperm morphology, in particular, the flagellar part showed several changes during activation in distilled water. Sperm motility of pike is inhibited due to high osmolality in the seminal plasma. Osmolality of activation medium affects the percentage of motile sperm and spermatozoa velocity due to changes in flagellar wave parameters.     

Steinernema ichnusae sp. n. (Nematoda: Steinernematidae) a new entomopathogenic nematode from Sardinia Island (Italy)

A new Steinernema species was isolated from three different sandy soil samples along the Platamona Beach, in the north-west coast of Sardinia Island (Italy). This new species is characterized by the following morphological characters: infective third-stage juvenile with a body length of 866 ± 61 (767-969) μm, distance from head to excretory pore of 63 ± 2.7 (59-68) μm, tail length of 81 ± 3.2 (76-89) μm, ratio E (%) 77 ± 3.4 (68-83); male tail with a mucron only in the second generation, spicule length of 66 ± 1.4 (64-67) μm and gubernaculum length of 44 ± 1.4 (43-46) μm in the first generation male; female of first generation with a slight vulval protrusion and ratio D (%) of 53 ± 4.0 (47-63). The new species differs distinctly from the related species (S. feltiae, S. kraussei, S. litorale, S. oregonense and S. cholashanense) in some morphometric values such as percentage of hyaline portion, ratios of gubernaculum/spicule length, spicule head length/width. The DNA analyses of the internal transcribed spacers and D2D3 regions show that the studied nematode isolates are a new species. Cross hybridisation tests with S. feltiae, S. kraussei, S. litorale, S. weiseri and S. oregonense showed that these species were reproductively isolated.     

A case of fatal hyperinfective strongyloidiasis with discovery of autoinfective filariform larvae in sputum

The autoinfective filariform larva of Strongyloides stercoralis causes hyperinfection in immunosuppressed hosts. Here we report on the case of a male patient who was admitted to the emergency room at Gwangju Veterans Hospital with a complaint of dyspnea, and who was receiving corticosteroid therapy for asthma. Many slender larvae of S. stercoralis with a notched tail were detected in Papanicolaou stained sputum. They measured 269 +/- 21.2 microm in length and 11 +/- 0.6 microm in width. The esophagus extended nearly half of the body length. The larvae were identified putatively as autoinfective third-stage filariform larvae, and their presence was fatal. The autoinfective filariform larva of S. stercoralis has not been previously reported in Korea.     

Fasciola gigantica: Cercarial shedding pattern from Lymnaea natalensis after long-term exposure to cadmium at different temperatures

Little is known about the response of Fasciola spp., inside its snail host to pollutant toxicity. The effect of long-term exposure of Lymnaea natalensis to 0.1 μg/l, 10 μg/l, or 100 μg/l cadmium (Cd) on the infection rates with F. gigantica, percentage of cercariae-shedding snails and cercariae shedding pattern was investigated. The snails were exposed to Cd for 7 days either pre-infection with single Fasciola miracidium or during the late pre-patent period of the infection. The possible interaction between metal exposure and acclimatization temperature was also studied in three ranges; 16-18 °C, 23-25 °C and 30-32 °C. Results clearly showed that host exposure to 100 μg/l Cd significantly affected the pattern of Fasciola development inside its snail host, and that the acclimatization temperature was a key factor affecting the role played by Cd. Pre-infection exposure to Cd caused a significant reduction in the infection rates where the effect was temperature-dependent. Post-infection exposure to Cd significantly increased the percentage of cercariae-shedding snails; the effect was temperature-independent. Interestingly, post-infection exposure significantly altered the differential cercarial output pattern, while no clear impact on the total was found. The alteration in the differential cercarial output was represented as a significantly higher proportion of floating metacercaria as compared to fixed ones and also higher proportion of dead cercariae which may directly reduce the transmission of Fasciola to the final host. The mechanisms of cadmium impact are briefly discussed.     

Lipid class dynamics during larval ontogeny of sea scallops, Placopecten magellanicus, in relation to metamorphic success and response to antibiotics

This study examines ontogenetic changes in lipid class composition in relation to survival and growth of five batches of sea scallops, Placopecten magellanicus, from egg to dissoconch stages. It also investigates the effects of antibiotic addition during metamorphosis to determine whether transient treatment during this critical period is effective in increasing metamorphic success. The top-performing larvae (growth rate: 4.7-5.0 μm day^− 1, dissoconch survival: 32-57%) were characterized by a pronounced increase in absolute (ng larva^− 1) and relative concentrations (wt.% of total lipid) of triacylglycerol (TAG) during the pre-metamorphic period, followed by utilization during metamorphosis (0.5-1.4 μg day^− 1 larva^− 1). In contrast, the low-performing scallops (growth rate: 3.6-4.5 μm day^− 1, dissoconch survival < 1%) exhibited a constant, low level of TAG. These results strongly suggest that the accumulation of TAG during the pre-metamorphic period is a good predictor of scallop performance as measured by survival to dissoconch stage. Antibiotic treatment enhanced absolute and relative TAG concentrations in pre-metamorphic scallops and resulted in higher post-metamorphic survival and TAG concentration than in non-treated controls. However, the proportion of dissoconch to live larvae at the end of the experiment was significantly lower with antibiotic treatment (8% vs. 20% in controls), thus resulting in comparable yields of dissoconch larvae ∼ 8% irrespective of treatment. A possible negative effect of antibiotic treatment on bacterial induction of settlement and metamorphosis of sea scallop larvae is suggested.     

Effect of Acanthocephala infection on the reproductive potential of crustacean intermediate hosts

The effect of a naturally acquired infection by three acanthocephalan parasites Dentitruncus truttae, Echinorhynchus truttae, and Polymorphus minutus on the reproductive potential of their intermediate host, Echinogammarus tibaldii (Amphipoda) from Lake Piediluco (Centre of Italy) was assessed. During May 2007, 1135 amphipods were collected from two different samplings and examined for larval helminths. Forty-five amphipods were infected and of those, 16 were infected with D. truttae (intensity = 1-3 larvae), 15 with E. truttae (intensity = 1-2 larvae), and 14 with P. minutus (intensity = 1 larva). The sex ratio was nearly 1:1 in all examined amphipods. One female infected with D. truttae contained six eggs in the brood pouch and another female infected with E. truttae contained five eggs. However, none of the eight female amphipods harbouring P. minutus larva contained eggs in their brood pouch. Uninfected females of the same size and body length as that of the infected females contained between 20 and 32 eggs. No acanthocephalan species were found to co-occur.     

Effect of light intensity on Opisthorchis viverrini cercarial shedding levels from Bithynia snails--a preliminary study

Opisthorchis viverrini requires Bithynia snails as the first intermediate host and cyprinid fish as the second intermediate host. Very low natural infection rates have been reported in Bithynia snails, but very high rates have been found in cyprinid fish in the same endemic region. This study investigated the effect of light intensity, the most important stimulus, on the quantity of O. viverrini cercariae shed from naturally infected Bithynia (Digoniostoma) siamensis goniomphalos snails. Snails were evaluated for cercariae output every hour after exposure to various light intensities for a total period of 7 h. The same infected snail was tested under different intensities of light: in the dark, and at 1000, 3000 and 5000 lx. The data showed that under exposure to 1000 and 3000 lx of light, the average percentage and number of cercariae released were higher than that exposed to 5000 lx during the first 2 h of the experiment. In contrast, under higher illumination (5000 lx) a longer time (6 h) was required to stimulate the peak emergence of cercariae. Darkness was not able to induce O. viverrini cercariae emergence. Among the three intensities of light, exposure at 1000 lx induced the highest average number of released cercariae per snail and the highest percentage of cercarial emergence within the first 2 h (125, 54.86%), followed by exposure at 3000 lx (69, 25.58%) and 5000 lx (12, 7.78%). The results suggest that the light intensity of 1000 lx for 2 h would be optimal for O. viverrini cercarial shedding from naturally infected B. (D.) siamensis goniomphalos snails.     

Molecular and ultrastructural characterization of Andreanna caspii n. gen., n. sp. (Microsporida: Amblyosporidae), a parasite of Ochlerotatus caspius (Diptera: Culicidae)

A new genus and species of microsporidia, Andreanna caspii n. gen., n. sp. is described from the mosquito, Ochlerotatus caspius (Pallas) based on ultrastructural morphology, developmental characteristics, and comparative sequence analyses of the small subunit (SSU) ribosomal DNA (rDNA). Parasite development is confined to fat body tissue and infected larvae appear swollen with dull white masses within the thorax and abdomen. Meronts have diplokaryotic nuclei and are delineated by a simple plasmalemma contiguous with the host cell cytoplasm. Merogony occurs by synchronous binary division followed by cytokinesis. Diplokaryotic sporonts undergo meiosis and synchronous nuclear division forming sporogonial plasmodia with two, four and eight nuclei enclosed within a persistent sporophorous vesicle. Cytokinesis of sporogonial plasmodia results in the formation of eight uninucleate spores. The episporontal space of early sporonts is filled with a homogeneous accumulation of electron dense granular inclusions and ovoid vesicles of various dimensions, transforming into an interwoven matrix during the initial phase of sporogenesis. Spores are oval, uninucleate and measure 4.8 ± 0.3 × 3.1 ± 0.4 μm (fixed). The spore wall is 260 μm thick with an irregular exospore consisting of two layers (150-170 μm) and a thinner endospore (90-100 μm). The anchoring disk is well developed and is contiguous with a lamellar polaroplast that occupies the anterior third of the spore and possess more narrow lamellae on the posterior end. The polar filament is gradually tapered and arranged in a single row consisting of six coils ranging from 180 to 150 μm in diameter. The posterior vacuole (posterosome) is moderately sized and filled with a matrix of moderate electron density. Phylogenetic analysis of SSU rDNA from A. caspii and 30 other species of microsporidia including 11 genera parasitic in mosquitoes using maximum parsimony, neighbor joining and maximum likelihood methods showed A. caspii to be a sister group to the clade containing all of the Amblyospora species, including Culicospora, Edhazardia and Intrapredatorus, as well as Culicosporella and Hyalinocysta thus providing strong support for establishment of Andreanna as a separate genus.     

Angiostrongylus cantonensis: experimental study on the susceptibility of apple snails, Pomacea canaliculata compared to Pila polita

Six groups (15 snails/group) of Pomacea canaliculata and Pila polita were infected orally with 0 (control), 200, 400, 800, 1600 and 3200 first-stage Angiostrongylus cantonensis larvae (L1). The respective mean+/-SD third stage larvae (L3) worm recovery 1-month post-infection (p.i.) for P. canaliculata was 0, 1.4+/-5.42 (0.7%), 0.13+/-0.35 (0.03%), 0.07+/-0.26 (0.009%), 0.07+/-0.26 (0.004%), 0, and for P. polita 0, 64.33+/-21.38 (32.25%), 115.36+/-36.82 (28.93%), 265.33+/-90.01 (33.27%), 471.33+/-92.98 (29.60%) and 849.00+/-243.23 (26.61%). The susceptibility of A. cantonensis in P. polita was dose-dependent (p<0.001). In the three groups (nine snails/group) of P. polita given 500 L1, we studied the distribution of L3 in the internal organs (i.e., foot, head+esophagus, kidney, albumin gland, mantle, intestine, digestive gland) and found the highest density after 1, 2 and 3 months p.i. in the mantle at 29.37%, 31.09% and 37.45%. The infection rate in P. canaliculata was too low to study distribution rates.     

Simultaneous detection of Giardia lamblia and Cryptosporidium parvum (oo)cysts in soil using immunomagnetic separation and direct fluorescent antibody staining

An improved approach for simultaneous detection of Cryptosporidium parvum and Giardia lamblia (oo)cysts in soil is described. Recoveries > 70% were obtained for concentrations > 55 and 21 (oo)cysts g ⁻¹ for C. parvum and G. lamblia, respectively. The limits of detection were determined to be < 5 (oo)cysts g ⁻¹ soil.     

Early development and pathogenesis of Lobatostoma manteri Rohde (Trematoda: Aspidogastrea)

The prosobranch Planaxis sulcatus is reported as a new natural host of Lobatostoma manteri at Heron Island, Great Barrier Reef. Planaxis sulcatus and Cerithium moniliferum were experimentally infected with large numbers of eggs. The larvae hatch in the stomach and migrate immediately along the ducts of the digestive gland into the digestive follicles. The larvae feed on the secretion and probably epithelial cells of the follicles. The acetabulum is used for adhesion to the epithelium and contributes to its erosion. In heavily infected snails, the digestive follicles disappear gradually and the larvae live in cavities lined by a flattened epithelium, parts of which show secretory activity. In snails dissected 47–49 and 65–66 days after infection, the cavities are fused, forming several large spaces which communicate with each other; only small parts of the epithelium are still secretory. Concentrations of amoebocytes occur in the walls of the digestive gland and in the wall between digestive gland and stomach of infected Planaxis. Some young worms were found in the stomach of Planaxis. No tissue reactions were seen around the stomach except in the wall between digestive gland and stomach. In Cerithium with 65–67 days old infection, the cavities contain much detritus and disintegrating cells, the epithelium is practically non-secretory and surrounded by loose connective tissue. In larvae with a body length of approximately 0·5–0·6 mm, the acetabulum begins to divide into alveoli and its anterior end grows forward; the anterior alveoli gradually increase in size and new alveoli are formed in the posterior undivided zone. In two specimens of approximately 1·3 mm body length, the whole adhesive disk was divided into half the number of alveoli usually found in adults. Allometric shifts during growth of the worms are analysed.     

Quantifying horizontal transmission of Nosema lymantriae, a microsporidian pathogen of the gypsy moth, Lymantria dispar (Lep., Lymantriidae) in field cage studies

Nosema lymantriae is a microsporidian pathogen of the gypsy moth, Lymantria dispar that has been documented to be at least partially responsible for the collapse of L. dispar outbreak populations in Europe. To quantify horizontal transmission of this pathogen under field conditions we performed caged-tree experiments that varied (1) the density of the pathogen through the introduction of laboratory-infected larvae, and (2) the total time that susceptible (test) larvae were exposed to these infected larvae. The time frame of the experiments extended from the early phase of colonization of the target tissues by the microsporidium to the onset of pathogen-induced mortality or pupation of test larvae. Upon termination of each experiment, the prevalence of infection in test larvae was evaluated. In the experiments performed over a range of pathogen densities, infection of test larvae increased with increasing density of inoculated larvae, from 14.2 ± 3.5% at density of 10 inoculated per 100 larvae to 36.7 ± 5.7% at 30 inoculated per 100 larvae. At higher densities, percent infection in test larvae appeared to level off (35.7 ± 5.5% at 50 inoculated per 100 larvae). When larval exposure to the pathogen was varied, transmission of N. lymantriae did not occur within the first 15 d post-inoculation (dpi) (11 d post-exposure of test larvae to inoculated larvae). We found the first infected test larvae in samples taken 20 dpi (16 d post-exposure). Transmission increased over time; in the cages sampled 25 dpi (21 d post-exposure), Nosema prevalence in test larvae ranged from 20.6% to 39.2%.