Production and chronobiology of emergence of the cercariae of Euparyphium albuferensis (Trematoda: Echinostomatidae)

The production and the chronobiology of emergence of the cercariae of Euparyphium albuferensis from Gyraulus chinensis experimentally infected with a single miracidium were established during 28 consecutive days from the first day of cercarial shedding. Moreover, the effect of a sudden change in light-dark cycling was investigated. Although the daily cercarial shedding rates show great variability, a progressive increase in cercarial production was observed in the first weeks of the cercarial shedding periods, probably in relation to the demography of intramolluscan larval stages. Under 12:12 light-dark cycling conditions, E. albuferensis cercariae emerged in the light, and the rhythm was circadian. The sudden change in the light-dark cycle resulted in corresponding alterations in emergence patterns, which shows that cercarial emergence was correlated to light-dark alternation. The ecological consequences of daily emergence by mobile cercariae whose target hosts are organisms that regularly occur in the same habitat with the molluscan emitting host, such as those of E. albuferensis, are discussed with reference to the hypotheses proposed to date, and an alternative hypothesis is proposed.     

The life cycle and transmission dynamics of the larval stages of Hypoderaeum conoideum

The morphology of the different larval stages and life cycle of Hypoderaeum conoideum (Trematoda: Echinostomatidae) are described. The freshwater snail species Lymnaea peregra (Gastropoda: Lymnaeidae) serves as the natural first intermediate host and this and L. corvus serve as experimental first intermediate hosts. These and other freshwater snails, such as Physella acuta and Gyraulus chinensis, in turn serve as second intermediate hosts. Adult worms were obtained from chicks and ducks, but not from rats, mice and golden hamsters. The morphology of the larval stages is compared with previous work on H. conoideum. Several aspects of the biology of the life history stages are described with emphasis on the transmission dynamics of the free-living stages. Differential suitability of the snail species that may act as first and/or second intermediate hosts is studied and discussed.     

Life-cycle of Euparyphium albuferensis n. sp. (Trematoda: Echinostomatidae) from rats in Spain

The morphology of the different life-history stages and life-cycle of Euparyphium albuferensis are described and drawn. The freshwater snail Gyraulus chinensis (Gastropoda: Planorbidae) serves as the natural and experimental first intermediate host. This and other freshwater snails, such as Lymnaea truncatula, L. peregra, L. palustris and Physa acuta, serve as second intermediate hosts. Adult worms, possessing 45 collar spines, were obtained naturally from Rattus norvegicus and R. rattus, and experimentally from albino rats, mice and golden hamsters. Chickens, ducks and pigeons were not suitable experimental definitive hosts. E. albuferensis differs from the most closely related species, E. murinum Tubangui, 1931, in its larger body, suckers and oesophageal measurements, in the distribution of vitelline follicles and in the morphology of the collar spines.     

Emergence of cercariae of Echinostoma caproni and Schistosoma mansoni from Biomphalaria glabrata under different laboratory conditions

Release of Echinostoma caproni cercariae and Schistosoma mansoni from experimentally infected Biomphalaria glabrata snails maintained under different laboratory conditions was studied. Infected snails were isolated individually for 1 h in Stender dishes containing 5 ml of artificial spring water and the number of cercariae released during this time was recorded. Of numerous conditions tested, the addition of lettuce, the use of water conditioned by B. glabrata snails and a temperature of 35 degrees C significantly increased the release of E. caproni cercariae. A significant increase in cercarial release of S. mansoni was seen only in cultures fed lettuce. A temperature of 12 degrees C caused a significant decrease in cercarial release of both E. caproni and S. mansoni. Increased snail activity associated with feeding behaviour was probably responsible for the enhanced cercarial sheds observed in this study.     

Survival of Theileria parva-infected adult Rhipicephalus appendiculatus under laboratory and quasi-natural conditions

Adult Rhipicephalus appendiculatus Muguga, having high or low intensities of Theileria parva Muguga infection in their salivary glands, were exposed to 20 °C and 85% relative humidity in the laboratory or quasi-natural conditions. Survival of the ticks and T. parva infections in their salivary glands was then monitored over a two year period. Ticks, having an average infection level of 2 infected acini per female, survived for up to 70 or 106 weeks after moulting under the laboratory or quasi-natural conditions respectively. Those having an infection level of 26 infected acini per female, survived for a similar duration except that those under quasi-natural conditions survived for a slightly shorter duration (102 weeks). Similarly, T. parva parasites survived for much longer periods under quasi-natural conditions than under the laboratory conditions. They survived for up to 38 or 78 weeks post salivary gland infection under the laboratory or quasi-natural conditions respectively in both categories of infection levels. There was apparently a density dependent relationship in T. parva survival, with a dramatic fall in infection occurring in ticks with high levels of infection between weeks 10 and 18 or weeks 38 and 46 post salivary gland infection in those exposed to laboratory or quasi-natural conditions before levelling off. This revised version was published online in July 2006 with corrections to the Cover Date.     

Persistence of human rhinovirus infectivity under diverse environmental conditions.

The persistence of human rhinovirus type 2 and type 14 infectivity was studied under various laboratory conditions designed to mimic those commonly found in the environment. The effects of temperature, ionic strength, protein content, and evaporation were compared. Both viruses were stable (less than 0.3-log decrease in titer) at 6 and 23 degrees c for 24 h in the liquid state regardless of salt or protein additives; a titer decrease of less than 1.0 log was noted at 37 degrees C. However, evaporation at 37 degrees C reduced virus infectivity by 3.2 to 4.5 logs in buffered water, an effect which could be significantly lessened by the addition of bovine serum albumin in saline (2.0- to 2.9-log decrease in titer). These studies support and extend observations by others that the human rhinoviruses retain sufficient infectivity after drying on hard surfaces to permit their transmission to susceptible persons upon contact.     

Survival of shelled acanthors of Moniliformis moniliformis (Acanthocephala) under laboratory conditions

Shelled acanthors (= eggs) of two isolates of Moniliformis moniliformis (Acanthocephala) were tested for their capacity to respond to a hatching stimulus in vitro and for their retention of infectivity to a natural intermediate host (Periplaneta americana). The shelled acanthors were stored for more than 120 weeks (Australian isolate) and for 104 weeks (Texan isolate) together with rat faeces in an incubator maintained at 22.2 +/- 0.1 degrees C. In both cases, infectivity to P. americana was shown to have been retained. In vitro tests of the hatching response were carried out on many occasions during this period of faecal storage. Shelled acanthors continued to respond and no differences were detected either between isolates or within an isolate through time.     

Survival and development of immature Aedes aegypti in artificial desiccating breeding sites under laboratory conditions

The ability of aquatic stages of Aedes aegypti (Diptera: Culicidae), key vector of dengue, chikungunya and Zika viruses, to survive in minimal residues of water has remained unstudied. We found that larvae eclosed from up to 50% of eggs set in permanent water films on water agar (at 1–2% agar), and some (≤9.5%) of these first‐instar larvae developed to second instar (L2). Mean larval survival was about 2 weeks; the longest periods of survival were 31.6 and 38.7 days in a water film and in free water controls, respectively. In another test, L3 moved in the films, with the greatest displacements of the larvae being up to 45 mm per day in the first 3 days; subsequent displacements in the water film were distinctly reduced during the next 20 days by which time all L3 had died. L3 did not detect or move in any directed manner in the film towards nearby small quantities of free water. Most pupae (≥93.7%) placed into water films or in water survived and developed to adults within 3 days. The unexpected survival of aquatic immature stages in desiccating breeding sites might be a reason for both the tenacious persistence of mosquito populations in seasons with infrequent rainfall and failures of classical control strategies against this important vector.     

Substratum preference of Philophthalmus sp. cercariae for cyst formation under natural and experimental conditions

Selection on parasites should favor adaptations that maximize the probability of transmission to the definitive host, such as the preference for and use of intermediate hosts or encystment substrata that are likely to be consumed by the definitive host. Eye flukes in the genus Philophthalmus are passed to their definitive avian host through the ingestion of metacercariae encysted on hard substrata. The life cycle of these parasites is generally well understood; however, there is almost no information on substratum use or preference of the cercariae of these parasites. In this study, we combine a survey of naturally occurring substrata with experimental, laboratory-based choice tests to determine the preferred substratum of Philophthalmus sp. and whether this preference is affected by the presence and density of pre-existing cysts. A concordance between natural and experimental data show a preference for the shells of multiple species of snail over other hard substrata that are common at the field site, including seaweed, other molluscs, and crustaceans. In addition, we found that cercariae preferred substrata with pre-existing cysts and that this preference seemed to increase with increasing cyst density. Such a preference should lead to an aggregated distribution of cysts among snail shells that may benefit the parasite by increasing the number of potential mates that become established in the definitive host. The identification of a preferred substratum also may help to identify potential definitive hosts that were previously unknown.     

Effect of selected chemical agents on longevity and infectivity of Schistosoma mansoni cercariae.

The effects of various chemical agents on longevity of the cercariae of Schistosoma mansoni Sambon, 1907, were investigated. The median lifetime of cercariae maintained in dechlorinated tap water (DTW) was 10.5 hr. Increasing concentrations of sodium chloride added to distilled water increased the median lifetime to an optimum of 10 hr at 0.01 M; higher salt levels decreased longevity. At this optimum sodium chloride level, concentrations of glucose between 0.003 M and 0.03 M enhanced median survival to 13–14 hr. Using Tris-HCl buffers the effects of pH and ionic strength were examined. Cercarial longevity increased from 3.5 to 25 hr as pH increased from 6.4 to 9.0, and 0.01 M Tris was superior to 0.001 M Tris at a given pH. The greatest median lifetime (26 hr) was obtained in 0.01 M Tris, pH 9.0. Infectivity of cercariae in Tris at this optimal pH was compared to that in DTW. Maintenance in DTW resulted in greater worm burdens in mice than did treatment with Tris. This suggested that factors which affect cercarial longevity may not influence infectivity in the same manner. The effects of rotenone, Antimycin A, dinitrophenol, and potassium cyanide on cercarial viability suggested the existence of a functioning terminal electron transport chain similar to that of mammalian systems.     

Persistence of human rhinovirus infectivity under diverse environmental conditions

The persistence of human rhinovirus type 2 and type 14 infectivity was studied under various laboratory conditions designed to mimic those commonly found in the environment. The effects of temperature, ionic strength, protein content, and evaporation were compared. Both viruses were stable (less than 0.3-log decrease in titer) at 6 and 23 degrees c for 24 h in the liquid state regardless of salt or protein additives; a titer decrease of less than 1.0 log was noted at 37 degrees C. However, evaporation at 37 degrees C reduced virus infectivity by 3.2 to 4.5 logs in buffered water, an effect which could be significantly lessened by the addition of bovine serum albumin in saline (2.0- to 2.9-log decrease in titer). These studies support and extend observations by others that the human rhinoviruses retain sufficient infectivity after drying on hard surfaces to permit their transmission to susceptible persons upon contact.     

Survival of coxsackievirus B3 under diverse environmental conditions.

The survival of coxsackievirus B3 was studied under various conditions of incubation. The comparative study demonstrated that coxsackievirus B3 was stable for 24h (less than 0.4-log decrease in titer) when suspended at neutral pH (6 or 23 degrees C) in the presence of 0.25% bovine serum albumin in saline regardless of whether the preparations were subjected to evaporation. Bovine serum albumin provided increased stability to the virus for each of the conditions tested. At 37 degrees C, evaporation greatly reduced the virus infectivity between 6 and 20 h of incubation. Nevertheless, coxsackievirus B3 was found to be stable for at least 24 h under conditions similar to those of a household environment, and its presence represents a potential biohazard to nonimmune persons. These data provide a rationale for using coxsackievirus B3 as a model for investigating the role of environmental surfaces in the transmission of enteroviral diseases.     

Survival of coxsackievirus B3 under diverse environmental conditions.

The survival of coxsackievirus B3 was studied under various conditions of incubation. The comparative study demonstrated that coxsackievirus B3 was stable for 24h (less than 0.4-log decrease in titer) when suspended at neutral pH (6 or 23 degrees C) in the presence of 0.25% bovine serum albumin in saline regardless of whether the preparations were subjected to evaporation. Bovine serum albumin provided increased stability to the virus for each of the conditions tested. At 37 degrees C, evaporation greatly reduced the virus infectivity between 6 and 20 h of incubation. Nevertheless, coxsackievirus B3 was found to be stable for at least 24 h under conditions similar to those of a household environment, and its presence represents a potential biohazard to nonimmune persons. These data provide a rationale for using coxsackievirus B3 as a model for investigating the role of environmental surfaces in the transmission of enteroviral diseases.     

Effects of exogenous glucose on survival and infectivity of Schistosoma mansoni cercariae

The effects of exogenous glucose in artificial spring water (ASW) were studied on the survival and infectivity of Schistosoma mansoni cercariae. The mean percent survival of cercariae maintained in 1% glucose in ASW for 36 and 48 hr was significantly greater than that of cercariae maintained identically in ASW. Cercariae maintained in ASW with or without glucose for 24 hr, fixed in neutral buffered formalin, and stained in Oil Red O, showed an accumulation of neutral lipid in the tail. Cercariae maintained as described above and stained in periodic acid-Schiff exhibited depleted glycogen, mainly from the tail. Cercariae maintained in ASW with glucose for 24 hr did not resynthesize glycogen. Cercariae maintained in ASW with glucose for 24 hr were as capable of infecting male FVBN202 mice as were freshly emerged cercariae, and increased the percent of worm recovery. Exogeneous glucose added to ASW prolonged the survival of S. mansoni cercariae and increased infectivity in terms of worm recovery.     

Cercarial chaetotaxy of Euparyphium albuferensis Esteban et al., 1997 (Trematoda: Echinostomatidae), with a review of some genera of the Echinostomatinae

Euparyphium albuferensis Esteban et al., 1997 (Trematoda: Echinostomatidae) is a parasite of the mollusc Gyraulus chinensis (D.) (Gastropoda: Planorbidae) and the mammals Rattus spp. in a nature park near Valencia, Spain. In the present work, the cercarial chaetotaxy is fully determined for specimens emitted by naturally infected G. chinensis. Of particular importance are the following sensillary arrangements: 5 CIV DL, 3 CIV CIV D at the dorso-cephalic level and 0 UV and 22–26 UD in the tail. Evidence is presented to the effect that these sensillary clusters are characteristic of the genus Euparyphium and allow an easy diagnosis, unlike in the case of the adult worms where there are sometimes few or no differences between Echinoparyphium and Echinostoma. A table synthesises the main data on the cercarial sensilla of Euparyphium spp. and the closely related genera Echinoparyphium, Echinostoma, Hypoderaeum, Moliniella, Neoacanthoparyphium, Paryphostomum and Isthmiophora. This shows clear variations in their respective numbers, from 3–5 in CIV DL, 2–4 in CIV D, 0–15 in UV and 19 to 57 in UD, according to the genus. These clusters appear adequate to identify, at the cercarial stage, the principal genera of the Echinostomatinae.     

Stability of Rauscher leukemia virus under certain laboratory conditions.

The stability of Rauscher leukemia virus (RLV) was investigated under certain laboratory conditions. The half life of the virus at 37 degrees was 7 hr, and considerably longer at lower temperatures. RNA dependent DNA polymerase activity was more stable than infectivity at all temperatures. Air dried virus had a half life of approximately 1 hr, but was rapidly inactivated by uv light or 70% alcohol.     

Daily emergence of Schistosoma mansoni and S. haematobium cercariae from naturally infected snails under field conditions

The daily emergence of Schistosoma mansoni and S. haematobium cercariae was investigated under field conditions. Intermediate host snails of both schistosome species were collected during the rainy season, cold dry season and warm dry season and kept separately in test tubes in habitat water. Shed cercariae were collected from each of the test tubes at two hourly intervals, transferred to Petri dishes and counted. Mice were exposed to these cercariae to establish the identity of the schistosome parasites. Peak shedding for both species was observed at 1100 h during the rainy and warm dry seasons and at 0900 h during the cold dry season. Shedding before 0900 h was found only for S. haematobium in the rainy season while shedding after 1700 h occurred only during this season at both species. Shedding observed during 1900 h observation period was in the low category for both species. No shedding was observed during the 2100 h observation period for any of the species and the investigation was discontinued after this period. Only S. haematobium ova were found in the exposed mice.     

Survival of directionally solidified B-lymphoblasts under various crystal growth conditions.

Reduction of temperature during freezing brings about two complex and interrelated phenomena: (1) crystal nucleation and subsequent growth processes and (2) change in biophysical properties of a biological system. The purpose of this investigation is to relate the morphology of the solid phase with the survival of a cell. To this end, B-lymphoblasts were exposed to directional solidification in phosphate-buffered saline + 0.05 M dimethyl sulfoxide. Directional solidification is a freezing technique which allows the morphology of the interface to be varied without varying the chemical history that a cell would experience during a constant cooling rate protocol. Results indicated that, for the range of experimental conditions tested, a maximum survival of approximately 78% could be achieved using a temperature gradient of 25(10)3 K/m and an interface velocity of 23(10)-6 m/s (cooling rate: 35 K/min). Survival dropped off sharply for freezing at faster cooling rates with little or no variation in survival for different crystal growth conditions. Survival at slower cooling rates decreased with decreasing cooling rate. It was observed, however, that the presence of secondary branches in the ice phase correlated with lower survival for a given cooling rate. These results indicated that not only is the redistribution of solute during freezing a potential source of damage during freezing but ice/cell interactions are also. Thus, the cooling rate alone may not be adequate to describe the freezing process.