In vitro efficacy of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> sensu lato against unfed <Emphasis Type="Italic">Amblyomma parvum</Emphasis> (Acari: Ixodidae)

Amblyomma parvum Aragão (Acari: Ixodidae) is a tick species found with wide distribution in the Neotropical region. Even though it is a wildlife-related tick, it is also a frequent parasite of domestic animals, is aggressive to human beings and may harbor pathogenic microorganisms. Therefore, it is a target species for control on domestic animals, particularly those at the rural–wildlife interface. Herein, the efficacy of two isolates (E9 and IBCB 425) of an entomopathogenic fungus, Metarhizium anisopliae sensu lato, already evaluated for ticks that parasitize domestic animals, was tested against unfed A. parvum adults. Both isolates displayed high acaricidal efficacy after immersion in fungal conidial suspensions for 5 min. Isolate E9 killed all ticks by the 7th day post-treatment, and isolate IBCB 425 did so by the 11th day. Tick mortality of 80 and 90% was achieved as early as the 3rd and 4th days, respectively, with both treatments. Thus, if a commercial M. anisopliae s.l. acaricide against domestic animal ticks is developed, it would also be effective against A. parvum.     

Threat level influences the use of venom in a scorpion species, <Emphasis Type="Italic">Tityus stigmurus</Emphasis> (Scorpiones,Buthidae)

Venom demands high metabolic costs of the organisms that produce it because it is comprised of a complex mixture of various toxins. Due to this high cost, venomous animals modulate the amount or type of venom used depending on factors such as size of prey or intensity of predation threat. This paper shows that Tityus stigmurus, a prevalent scorpion species in the urban environment in the Northeast of Brazil, modulates its venom in response to different levels of stimuli. Sixty animals were collected in Vitória de Santo Antão, Pernambuco. The animals were subjected to either high or low levels of threats. During the tests, the animals were gently touched five times on the mesosoma with forceps at an interval of 5 s (high threat) or 5 min (low threat). The response varied significantly between intensity levels, with the animals exposed to low threat stinging in 70% of the observations and releasing a clear venom. In contrast, individuals subjected to a high level of threat stung in 83% of the observations and released a milky venom. Our results suggest that T. stigmurus reacts differently depending on the stimulus level. When the threat was considered high, the animal reacted more aggressively. Our results support the assumption that milky venom is only used when the animal is highly stressed because this venom represents higher metabolic costs than the production of clear venom.     

Common cellular events occur during wound healing and organ regeneration in the sea cucumber <Emphasis Type="Italic">Holothuria glaberrima</Emphasis>

Background  

All animals possess some type of tissue repair mechanism. In some species, the capacity to repair tissues is limited to the healing of wounds. Other species, such as echinoderms, posses a striking repair capability that can include the replacement of entire organs. It has been reported that some mechanisms, namely extracellular matrix remodeling, appear to occur in most repair processes. However, it remains unclear to what extent the process of organ regeneration, particularly in animals where loss and regeneration of complex structures is a programmed natural event, is similar to wound healing. We have now used the sea cucumber Holothuria glaberrima to address this question.     

Radio-controlled up-net enclosure to capture free-ranging Alpine chamois <Emphasis Type="Italic">Rupicapra rupicapra</Emphasis>

We used an enclosure trap with a lifting net to capture Alpine chamois (Rupicapra rupicapra). The trap was activated by remote radio-controlled electromagnets powered by photovoltaic modules. The up-net trap had considerable advantages over mechanical methods described in the literature for the capture of chamois. During 36 capture sessions, we captured 50 chamois, or 1.39 animals per session. Capture success was 96.2% of the average of 1.4 animals that entered the trap during a capture session. Mortality was 2%, and another 2% of the captured chamois was injured. There were no known postrelease capture-induced pathologies, and the capture effort was 1.7 man-days per chamois. The trap allowed to select specific target animals and to capture sex–age classes (particularly kids and their mothers) that are normally difficult or impossible to capture. It could be set off from a distance; it only required about 4 h to assemble and could be operated by as few as two people.     

Comparison of Mercury Contamination in Live and Dead Dolphins from a Newly Described Species,Tursiops australis

Globally it is estimated that up to 37% of all marine mammals are at a risk of extinction, due in particular to human impacts, including coastal pollution. Dolphins are known to be at risk from anthropogenic contaminants due to their longevity and high trophic position. While it is known that beach-cast animals are often high in contaminants, it has not been possible to determine whether levels may also be high in live animals from the same populations. In this paper we quantitatively assess mercury contamination in the two main populations of a newly described dolphin species from south eastern Australia, Tursiops australis. This species appear to be limited to coastal waters in close proximity to a major urban centre, and as such is likely to be vulnerable to anthropogenic pollution. For the first time, we were able to compare blubber mercury concentrations from biopsy samples of live individuals and necropsies of beach-cast animals and show that beach-cast animals were highly contaminated with mercury, at almost three times the levels found in live animals. Levels in live animals were also high, and are attributable to chronic low dose exposure to mercury from the dolphin''s diet. Measurable levels of mercury were found in a number of important prey fish species. This illustrates the potential for low dose toxins in the environment to pass through marine food webs and potentially contribute to marine mammal deaths. This study demonstrates the potential use of blubber from biopsy samples to make inferences about the health of dolphins exposed to mercury.     

PCR-Denaturing Gradient Gel Electrophoresis and Two Feces Antigen Tests for Detection of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> in Mice

PCR-denaturing Gradient Gel Electrophoresis (PCR-DGGE), a method suitable for the detection of microbial species in complex ecosystems, was evaluated for the detection and identification of Helicobacter spp. in feces and stomach tissue of mice. Two commercially available stool antigen tests for clinical diagnostics in humans were also evaluated in the C57B1/6 mouse model of H. pylori infection. PCR-DGGE detected only Helicobacter ganmani in feces from H. pylori-infected as well as control animals, whereas in stomach specimens it demonstrated the presence of H. pylori in challenged and H. ganmani in control animals. Hence, the method detected DNA only of the predominant Helicobacter spp., which was also shown in cell dilution experiments. The Amplified IDEIA Hp StAR feces antigen test detected H. pylori in feces from all infected animals and generated no false-positive results, whereas the Premier Platinum HpSA-test also detected H. pylori in all infected animals but generated false-positive or equivocal results in 50% of the control animals. Premier Platinum HpSA, as opposed to Hp StAR, cross-reacted with non-pylori Helicobacter spp. in vitro.Received: 21 August 2002 / Accepted: 6 December 2002     

BIOCHEMICAL CHARACTERIZATION OF β-N-OXALYL-l-α,β-DIAMINOPROPIONIC ACID, THE LATHYRUS SATIVUS NEUROTOXIN AS AN EXCITANT AMINO ACID

Abstract— β- N -Oxalyl- l -α,β-diaminopropionic acid (ODAP), the toxin isolated from the seeds of Luthyrus sativus produces head retraction, tremors and convulsions when injected into a variety of experimental animals. In 12-day-old rats, it has been found that the convulsive behaviour is accompanied by profound biochemical changes in the brain. The brain homogenates prepared from ODAP injected animals show a higher rate of respiration. There is a decrease in the brain glucose, glycogen, ATP, phosphocreatine and acetylcholine levels of the convulsing animals. The inorganic phosphate, lactic acid and acetylcholineesterase levels increase. These results establish that ODAP is a typical convulsant.     

Chytrid infection reduces thoracic beat and heart rate of <Emphasis Type="Italic">Daphnia</Emphasis><Emphasis Type="Italic">pulicaria</Emphasis>

Zooplankters are hosts to numerous endo- and ectoparasites, some of which have dramatic impacts on their hosts. Epizootics on zooplankton are probably more widespread in lake systems than it is currently known, and few studies have explored the direct and indirect importance of parasitism in aquatic food webs. In addition, our understanding of the sublethal effects of parasitic infections on host organisms and populations is limited. We used a novel electro-chemical based technique to measure in the outflow of the feeding current changes in the beat rate of the thoracic appendages in female Daphnia pulicaria. We observed simultaneously the heart rates and compared chytrid infected animals with uninfected gravid and non-gravid ones. We found in uninfected animals a thoracic beat rate of 3.81 ± 018 Hz and a heart rate of 4.67 ± 0.42 Hz. Gravid daphnids had a 14% lower thoracic beat rate (3.27 ± 0.30 Hz) than non-gravid females while the heart rate did not significantly differ (4.48 ± 0.28 Hz). In contrast, infected animals showed a 22% lower thoracic beat rate (2.96 ± 0.47 Hz) and a 36% lower heart rate (2.98 ± 0.5 Hz) when compared with uninfected non-gravid females. We discuss the ways Daphnia are affected by Polycaryum leave infections on the individual and population level.     

Identification of tomato plant as a novel host model for <Emphasis Type="Italic">Burkholderia pseudomallei</Emphasis>

Background  

Burkholderia pseudomallei is the causative agent for melioidosis, a disease with significant mortality and morbidity in endemic regions. Its versatility as a pathogen is reflected in its relatively huge 7.24 Mb genome and the presence of many virulence factors including three Type Three Secretion Systems known as T3SS1, T3SS2 and T3SS3. Besides being a human pathogen, it is able to infect and cause disease in many different animals and alternative hosts such as C. elegans.     

Trade-offs between microhabitat selection and physiological plasticity in the Antarctic springtail, <Emphasis Type="Italic">Cryptopygus antarcticus</Emphasis> (Willem)

In the maritime Antarctic, terrestrial arthropods have recourse to two strategies to mitigate low summer temperatures: (1) physiological plasticity and (2) avoidance via microhabitat insulation. This study investigated the interaction between these strategies in the springtail, Cryptopygus antarcticus, established in situ within contrasting microcosms (buffered vs. exposed) and within two sets of habitat simulations (wet vs. dry) over diurnal scales through the Antarctic summer. Significant differences were found in the cold hardiness of springtails sampled simultaneously from each microcosm. Exposed animals showed greater plasticity in the “true” austral summer, but as field temperatures declined preceding the onset of winter, buffered animals showed greater resilience. Overall, water was found to inhibit the buffering effect of moss and there was a significantly greater discrimination between buffered and exposed microcosms in the dry treatment. Analysis of microhabitat temperatures indicate that it is thermal variability not lower temperature that is responsible for the greater plasticity of exposed animals.     

Evidence for the Placental Transfer of <Superscript>3</Superscript>H-Acetyl-Melatonin

DEVELOPMENTAL studies on the enzymes which form melatonin, the putative pineal hormone, indicate that it probably is not formed in rats before the tenth day of life^1–3. Melatonin could nevertheless arise in animals younger than 10 days of age by transfer from the mother and we now describe how we have investigated this possibility by determining whether ³H-acetyl-melatonin injected into a pregnant rat could be detected in its foetuses.     

<Emphasis Type="Italic">Entamoeba histolytica</Emphasis> and <Emphasis Type="Italic">E. dispar</Emphasis> infections in captive macaques (<Emphasis Type="Italic">Macaca fascicularis</Emphasis>) in the Philippines

Entamoeba histolytica is a protozoan parasite that infects man and animals. This parasite has a global distribution and the disease it causes is usually characterized by diarrhea. In order to detect the parasite, it is necessary to differentiate it from Entamoeba dispar. E. dispar appears morphologically similar to E. histolytica but does not cause disease and tissue invasion. This study reports on the prevalence of E. histolytica and E. dispar among captive macaques in a primate facility in the Philippines. PCR was used to correctly identify both Entamoeba species. Indirect fluorescent antibody test (IFAT) was also performed to determine the seroprevalence of amebiasis in the captive macaques. Based on PCR targeting of the peroxiredoxin gene, of the 96 stool samples collected, 23 (24%) contained E. histolytica while 32 (33%) contained E. dispar. IFAT revealed 26 (27%) serum samples positive for antibodies against E. histolytica. Sequence analysis of the 18S rRNA gene showed that the 23 E. histolytica isolates were identical to human E. histolytica isolates deposited in the GenBank and not Entamoeba nuttalli as found in macaques in other recent reports. The Philippines is a major exporter of monkeys for biomedical research purposes, so screening animals before transporting them to other locations lessens the risk of spreading zoonoses to a wider area. This is the first report of the molecular detection of E. histolytica and E. dispar among macaques in the Philippines. This study complements the limited information available on the animal hosts of E. histolytica in the Philippines.     

Implementing long‐term baselines into primate tool‐use studies

Studies on primate tool‐use often involve the use of baseline conditions, as they allow for the examination of any differences in the subjects' behavior before and after the introduction of a tool‐use task. While these baseline conditions can be powerful for identifying the relative contributions of individual and social learning for the acquisition of tool‐use behaviors in naïve (usually captive) subjects, many have criticized them for being too short, and not allowing enough time for the behavior to develop spontaneously. Furthermore, some wild tool‐use behaviors such as chimpanzee nut‐cracking require animals to manipulate and familiarize themselves with the materials of the behavior within a “sensitive learning period” before it develops later on in life. One solution to this problem is to implement long‐term baselines, in which, with collaboration with zoological institutions, the materials of the behavior are left in the enclosure for an extended period. The keepers would then be asked not to demonstrate or train the animals in the target behavior, but to report back to the researchers if they observe the behavior emerge during this extended period. Alongside keeper reports, video cameras could be installed in the enclosure to minimize the chance of false negatives and to allow for coding and inter‐rater reliability to be carried out on the videos. These long‐term baselines therefore provide extended enrichment opportunities for the animals, alongside allowing the zoological institution to publicize their involvement with the study and guests to observe animals interacting with different testing apparatuses and tools. Finally, long‐term baselines can provide invaluable insight on the individual and social learning abilities of primates as well as the potential development stages and sensitive learning periods required for specific behaviors.     

An extracellular Leptospira interrogans leucine‐rich repeat protein binds human E‐ and VE‐cadherins

Pathogenic Leptospira bacteria are the causative agents of leptospirosis, a zoonotic disease affecting animals and humans worldwide. These pathogenic species have the ability to rapidly cross host tissue barriers by a yet unknown mechanism. A comparative analysis of pathogens and saprophytes revealed a higher abundance of genes encoding proteins with leucine‐rich repeat (LRR) domains in the genomes of pathogens. In other bacterial pathogens, proteins with LRR domains have been shown to be involved in mediating host cell attachment and invasion. One protein from the pathogenic species Leptospira interrogans, LIC10831, has been previously analysed via X‐ray crystallography, with findings suggesting it may be an important bacterial adhesin. Herein we show that LIC10831 elicits an antibody response in infected animals, is actively secreted by the bacterium, and binds human E‐ and VE‐cadherins. These results provide biochemical and cellular evidences of LRR protein‐mediated host–pathogen interactions and identify a new multireceptor binding protein from this infectious Leptospira species.     

Comparative study of the heterophil phagocytic function in young and old ring doves (<Emphasis Type="Italic">Streptopelia risoria</Emphasis>) and its relationship with melatonin levels

A functional connection between the pineal gland (via the hormone melatonin) and the immune system has been suggested. In our previous results in the ring dove, we observed diurnal oscillations in the levels of this neurohormone in young animals and a decline in its plasma levels with advancing age (which is accompanied by the absence of diurnal rhythm). We also noted enhanced phagocytic activity of heterophils from old animals after in vitro incubation with both physiological and pharmacological doses of melatonin. Here, we evaluate the functional capacity of ring dove (Streptopelia risoria) heterophils in young (2 years of age) and old (8 years and more) animals at different times of day (0:00, 10:00 and 16:00, the times when the maximum, minimum, and mean values, respectively, of melatonin levels are observed in young animals). The phagocytic capacities for the ingestion of latex beads and Candida albicans were evaluated, as well as the oxidative metabolism which accompanies phagocytosis. At all three times of day studied, the heterophil phagocytic function with both latex and C. albicans was significantly greater in the young than in the old animals, and in the young animal cells it was significantly higher at 0:00. In addition, in the presence of latex beads, there was a significant decline at 10:00 and 0:00 of superoxide anion levels in the young animals relative to the old. In the young animals, there was a decline at 0:00 in comparison with both 10:00 and 16:00, and in the old animals there was a decline at both 0:00 and 16:00 compared with 10:00. These results could be due, at least in part, to the absence of a diurnal rhythm of melatonin in old animals, and to an enhancing effect of that hormone on young animals heterophil phagocytic function, which would also neutralize the oxidative stress deriving from this immune function.Abbreviations PBS phosphate-buffered saline - MIF macrophage migration-inhibition factor - PI phagocytosis index - PP phagocytosis percentage - PE phagocytosis efficiencyCommunicated by G. Heldmaier     

Seasonal Changes in Microsomal Fraction Enriched with Na,K-ATPase from Kidneys of the Ground Squirrel <Emphasis Type="Italic">Spermophilus undulatus</Emphasis>

The Na,K-ATPase activity in microsomal fraction isolated from kidneys of winter hibernating ground squirrels was found to be 1.8–2.0-fold lower than that in active animals in summer. This is partially connected with a decrease in Na,K-ATPase protein content in these preparations (by 25%). Using antibodies to different isoforms of Na,K-ATPase α-subunit and analysis of enzyme inhibition by ouabain, it was found that the decrease in Na,K-ATPase activity during hibernation is not connected with change in isoenzyme composition. Seasonal changes of Na,K-ATPase a-subunit phosphory- lation level by endogenous protein kinases were not found. Proteins which could be potential regulators of Na,K-ATPase activity were not found among phosphorylated proteins of the microsomes. Analysis of the composition and properties of the lipid phase of microsomes showed that the total level of unsaturation of fatty acids and the lipid/protein ratio are not changed significantly during hibernation, whereas the cholesterol content in preparations from kidneys of hibernating ground squirrels is approximately twice higher than that in preparations from kidneys of active animals. However, using spin and fluorescent probes it was shown that this difference in cholesterol content does not affect the integral membrane micro-viscosity of microsomes. Using the cross-linking agent cupric phenanthroline, it was shown that Na,K-ATPase in mem- branes of microsomes from kidneys of hibernating ground squirrels is present in more aggregated state in comparison with membranes of microsomes from kidneys of active animals. We suggest that the decrease in Na,K-ATPase activity in kidneys of ground squirrels during hibernation is mainly connected with the aggregation of proteins in plasma membrane.     

The biological function of gulls' attraction towards predators

Many species of birds and mammals are attracted towards some of their natural predators, often without overt aggression. The biological function of this predator attraction was studied in a colony of herring gulls and lesser black-backed gulls, with the aid of predator models. More birds flocked above the predator model when it was presented together with a dead gull; also the birds did not land as close to a predator with a dead gull as they did to a predator alone. Having seen a predator with a dead gull had a clear effect on the distance at which birds later avoided that particular predator on the ground when it was presented to them alone. It is suggested that predator-attraction enables animals to collect information about a potential enemy; in this information the experience of conspecifics with the predator is taken into account; subsequent behaviour of the animals depends partly on this information. The increase in the avoidance distance with respect to a predator with a dead bird may be an adaptation to carnivores' habit of surplus killing.     

Reduced P2x<Subscript>2</Subscript> receptor-mediated regulation of endocochlear potential in the ageing mouse cochlea

Extracellular adenosine triphosphate (ATP) has profound effects on the cochlea, including an effect on the regulation of the endocochlear potential (EP). Noise-induced release of ATP into the endolymph activates a shunt conductance mediated by P2X₂ receptors in tissues lining the endolymphatic compartment, which reduces the EP and, consequentially, hearing sensitivity. This may be a mechanism of adaptation or protection from high sound levels. As inaction of such a process could contribute to hearing loss, this study examined whether the action of ATP on EP changes with age and noise exposure in the mouse. The EP and the endolymphatic compartment resistance (CoPR) were measured in mice (CBA/CaJ) aged between 3 and 15 months. The EP and CoPR declined slightly with age with an associated small, but significant, reduction in auditory brainstem response thresholds. ATP (100–1,000 μM) microinjected into the endolymphatic compartment caused a dose-dependent decline in EP correlated to a similar decrease in CoPR. This was blocked by pyridoxal-phosphate-6-azophenyl-2′,4′-disulfonate, consistent with a P2X₂ receptor-mediated shunt conductance. There was no substantial difference in the ATP response with age. Noise exposure (octave-band noise 80–100 decibels sound pressure level (dBSPL), 48 h) in young animals induced an upregulation of the P2X₂ receptor expression in the organ of Corti and spiral limbus, most noticeably with the 90-dB exposure. This did not occur in the aged animals except following exposure at 90 dBSPL. The EP response to ATP was muted in the noise-exposed aged animals except following the 90-dB exposure. These findings provide some evidence that the adaptive response of the cochlea to noise may be reduced in older animals, and it is speculated that this could increase their susceptibility to noise-induced injury.     

Habitat utilization by an invasive herbivorous fish (<Emphasis Type="Italic">Siganus rivulatus</Emphasis>) in its native and invaded range

Movement is essential for understanding the distribution and abundance of animals. While it has been suggested that invasion success can be facilitated by species’ ability to adapt to novel environments, direct comparisons of movement patterns between native and invaded ranges of animals in their natural habitat are rare. The rivulated rabbitfish Siganus rivulatus was introduced from the Red Sea into the Mediterranean, where it is now found in extremely high abundances, and has overgrazed the coastal marine ecosystem in many locations. Through a continuous acoustic tracking system, we found that the movement of S. rivulatus individuals at a Mediterranean site differed substantially from those at a Red Sea site, with individuals in the Mediterranean having larger overall home ranges and lower site fidelity. However, no variation between sites was found in daily home range sizes. Results show that at the Mediterranean site S. rivulatus individuals have a larger spatial footprint, which may contribute to their impact and ability to expand their distribution. This study demonstrates a potential shift in individual movement of a marine invasive species between its native and invaded range, and highlights the role of movement in understanding biological invasions.