Digestive enzyme activities in herbivorous and carnivorous prickleback fishes (Teleostei: Stichaeidae): ontogenetic, dietary, and phylogenetic effects

We measured the activities of eight digestive enzymes in four species of herbivorous and carnivorous prickleback fishes and determined the effects of ontogeny, diet, and phylogeny on these enzyme activities. Of the four species, Cebidichthys violaceus and Xiphister mucosus shift to a more herbivorous diet as they grow (> or =45 mm SL [standard length]), whereas Xiphister atropurpureus and Anoplarchus purpurescens remain carnivores throughout life. Digestive enzyme activities of small (30-40 mm SL) carnivorous juveniles were compared with those of larger (60-75 mm SL) wild-caught juveniles that had consumed a natural diet and larger (60-75 mm SL) juveniles raised on a high-protein animal diet. Cebidichthys violaceus and both species of Xiphister showed ontogenetic changes in digestive enzyme activities, whereas A. purpurescens did not. Despite dietary differences between X. atropurpureus and X. mucosus, these sister taxa displayed the most similar digestive enzyme activities from ontogenetic and dietary perspectives (high alpha-amylase and lipase and low trypsin and aminopeptidase activities), and both were more similar to C. violaceus, a member of the same largely herbivorous clade, than either was to A. purpurescens, a member of an adjacent, carnivorous clade. The results support the hypothesis that phylogeny influences digestive enzyme activities in these fishes. Anoplarchus purpurescens, a carnivore with a diverse diet, showed great plasticity in enzyme activity, especially trypsin and aminopeptidase, which were elevated in this species to the highest level among the four species after consuming the high-protein diet. These results support the hypothesis that fishes with relatively broad diets can modulate digestive enzyme activities in response to changes in dietary composition.     

Insectivore to frugivore: ontogenetic changes in gut morphology and digestive enzyme activity in the characid fish Brycon guatemalensis from Costa Rican rain forest streams

Brycon guatemalensis , a Neotropical characid fish, consumes an entirely terrestrial diet, shifting from eating insects as juveniles to fruits and leaves as adults. Juvenile and larger‐sized fish collected in the Rio Puerto Viejo at the La Selva Biological Station in Costa Rica were studied to test the hypotheses that, with ontogeny, (1) relative gut length increases, (2) pyloric caeca arrangement and number remain unchanged and (3) pepsin, trypsin and lipase activities decrease, while α‐amylase activity increases. These hypotheses were mainly supported in that larger fish had longer guts, unchanged pyloric caeca arrangement but fewer caeca, and, at both environmental and standard temperatures for the enzyme assays, lower pepsin and trypsin activities but higher α‐amylase activities than the juveniles. Only lipase, among the digestive enzymes, exhibited the unexpected outcome of either not differing significantly in activity (per g of tissue) between juveniles and larger fish or being significantly higher (per mg of protein) in larger fish. The overall results support the view that B. guatemalensis is specialized morphologically and biochemically to function first as a carnivore and then as a herbivore during its life history.     

Relationships among four genera of mojarras (Teleostei: Perciformes: Gerreidae) from the western Atlantic and their tentative placement among percomorph fishes

A phylogenetic study of the percoid family Gerreidae at both lower and higher taxonomic levels is presented based on DNA sequence data of four genes: mitochondrial 12S and 16S, and nuclear genes rhodopsin and recombination activating gene 1 (RAG1). The taxonomic sampling includes four genera of Gerreidae from the western Atlantic, 39 additional percomorph representatives and two outgroups. Phylogenetic results confirm the monophyly of the Gerreidae and suggest that the family is divided into two sub-groups ( Diapterus auratus plus Eugerres plumieri and Eucinostomus gula plus Gerres cinereus ), which correspond to two previously defined taxonomic assemblages characterized by the shape of the preoperculum. Gerreids are placed at an intermediate position in the percomorph tree between two basal clades (L and Q) and a terminal clade N (grouping tetraodontiforms, acanthuroids, lophiiforms, caproids and several percoids). In addition, topology tests indicate that two traditional assemblages, Labroidei (seven representatives sampled) and Percoidei (22 representatives sampled) are not natural groups. Labrids and scarids appear to be more closely related to gerreids and to the members of clade N than to any other basal percomorphs, including their labroid 'allies' sampled in this study, Embiotocidae, Pomacentridae and Cichlidae, which are all nested within clade Q that also includes atherinomorphs, mugiliforms and Chandidae. The percoid taxa included in this study are widely distributed among various percomorph lineages. The percomorph phylogeny obtained is highly congruent with results from recent molecular studies.     

The complete mitochondrial DNA (mtDNA) of the donkey and mtDNA comparisons among four closely related mammalian species-pairs

The nucleotide sequence of the complete mitochondrial genome of the donkey, Equus asinus, was determined. The length of the molecule is 16,670 bp. The length, however, is not absolute due to pronounced heteroplasmy caused by variable numbers of two types of repetitive motifs in the control region. The sequence of the repeats is (a) 5′-CACACCCA and (b) 5′-TGCGCGCA, respectively. The order of (a) and (b) can be expressed as {n[2(a)+(b)]+m(a)}. In 32 different clones analyzed the number of n and m ranged from 0 to 9 and 1 to 7. The two rRNA genes, the 13 peptide-coding genes, and the 22 tRNA genes of the donkey and the horse, Equus caballus, were compared in detail. Total nucleotide difference outside the control region was 6.9%. Nucleotide difference between peptide-coding genes ranged from 6.4% to 9.4% with a mean of 8.0%. In the inferred protein sequences of the 13 peptide-coding genes the amino acid difference was 0.2–8.8%, and the mean for the 13 concatenated amino acid sequences was 1.9%. In the 22 tRNA genes, the mean difference was 3.5%, and that in the two rRNA genes was 4.1%. The mtDNA differences between the donkey and the horse suggest that the evolutionary separation of the two species occurred ≈9 million years ago. Analyses of differences among the mtDNAs of three other species-pairs, harbor seal/grey seal, fin whale/blue whale, and Homo/common chimpanzee, showed that the relative evolutionary rate of individual peptide-coding genes varies among different species-pairs and modes of comparison. The findings show that the superimposition of sequence data of one lineage for resolving and dating evolutionary divergences of other lineages should be performed with caution unless based on comprehensive data. Received: 15 October 1995 / Accepted: 15 April 1996     

New ecological information onScytalina cerdale (Pisces: Scytalinidae) from a central California rocky intertidal zone

Synopsis New information regarding the ecology ofScytalina cerdale was obtained over a four year period as a consequence of a long-term marine ecological study at the Diablo Canyon Power Plant (DCPP), San Luis Obispo County, California. Twenty intertidal fish surveys were conducted at approximately quarterly intervals, between March 1979 and June 1983, at three separate rocky shore locations (stations). During each survey, a total of 108 square meters (36 m² per station) was searched for fish during periods of low tide. A total of 280S. cerdale were collected, identified, measured, and released back into the same 4 m² area, from the same intertidal station (Diablo Cove), throughout the study period. This limited intertidal occurrence most likely reflectsS. cerdale microhabitat requirements; a combination of intertidal elevation (mean = +0.3 MLLW), substratum specificity (loose gravel, 5–10mm size range, overlying a base of sand and shell fragments), and degree of wave exposure (semi-protected). Throughout the study,S. cerdale was seasonally more abundant during summer months (June through August) and less abundant during winter months (November through February). The only exception to this abundance trend followed the 1982 winter storms, which coincided with an El Niño event, whenS. cerdale abundance uncharacteristically dropped during the subsequent 1983 spring and summer surveys. Mean total lengths did not vary greatly, reflecting the absence of early juvenile fishes, and relatively high mean fish densities (3.5 fish per m² for 20 surveys) were recorded. Qualitative comparisons betweenS. cerdale abundance and seasonal changes in water temperature indicated an inverse relationship. Gravid females, demersal egg masses, and early juvenile individuals were never observed during the four years of the study. This suggests that unlike most other intertidal fishes, particularly other blennioids,S. cerdale may not utilize the intertidal zone for reproductive and recruitment purposes; Diablo Canyon is the most southerly reported distribution for this intertidal species, and therefore, may not entirely represent the species' biology as a whole.     

Assimilation of carbon from dietary and nondietary macroalgae by a temperate-zone intertidal fish, Cebidichthys violaceus (Girard) (Teleostei: Stichaeidae)

Force-feeding experiments were performed to measure the assimilation of carbon by the herbivorous stichaeid fish Cebidichthys violaceus (Girard) from macroalgae either regularly consumed or avoided by the fish. Dietary species were the green alga Ulva lobata (Kütz.) S. & G. and the red algae Porphyraperforata J. Ag. and Iridaea flaccida (S. & G.) Silva while the nondietary species were the brown algae Macrocystis integrifolia Bory and Fucus distichus (de la Pyl.) Pow. ¹⁴C was found in five body compartments (stomach, intestine, liver, carcass and skin) of fish fed labeled portions of Ulva lobata, Porphyra perforata and Macrocystis integrifolia. Approximately equal amounts of ¹⁴C were assimilated from diatom-free thalli of Ulva lobata that were either untreated or treated with antibiotics to remove bacterial populations. Labeled carbon was also assimilated from bacteria- and diatom-free thalli of Iridaea flaccida, Macrocystis integrifolia and Fucus distichus. Assimilation efficiencies of unlabeled carbon (determined from CHN analyses of food and feces) varied widely among the dietary and nondietary algae. The markedly lower levels of carbon assimilation from the brown algae may be due to reduced digestibility of their carbohydrates and, for F. distichus, the presence of secondary compounds. The results of this study clearly show that a temperate marine fish can assimilate carbon from taxonomically diverse algae largely free of epibionts.     

Isozyme criteria in the testing of phyletic relationships between species of Gobius and related eastern Atlantic-Mediterranean genera (Teleostei: Gobüdae)

Horizontal starch gel electrophoresis was employed to investigate allozyme variation at 19 isozyme loci, in nine closely-related Atlantic-Mediterranean gobiid fish species, to assess genetic relationships, and to compare this with a phyletic hypothesis based on morphological apo-morphies. The species examined were Gobius niger, G. auratus, G. cruentatus, G. paganellus, Mauligobius maderensis, Zosterisessor ophiocephalus, Thorogobius ephippiatus, Padogobius martensii and P. nigricans . Various phenetic and cladistic analyses were performed on isozyme and morphological data. The phenetic and cladistic results from morphological data, and the cladistic results from isozyme data, were largely comparable, with P. martensü and P. nigricans forming a sister group to all the other taxa, species of Cobius forming a crown group within the latter, and stem lines formed by Mauligobius, Zosterisessor and Thorogobius , respectively. The isozymic estimates of genetic distance differed from these chiefly in the relative positions of Zosterisessor and G. auratus , which appear less similar to other species of Gohius. G. auratus shows many alleles which are unique within the genus Gobius ; possible explanations for this are put forward. The position of nigricans as congeneric with Padogobius martensü is confirmed by both phenetic and cladistic analyses of isozyme data.     

Morphological comparison and DNA barcoding of four closely related species in the genera Misgurnus and Paramisgurnus (Cypriniformes: Cobitidae)

To evaluate the feasibility of morphological and genetic identification of the closely related species in the genera Misgurnus and Paramisgurnus, the morphological characters of four species in these genera and DNA barcoding of five loaches (P. dabryanus, M. anguillicaudatus, M. bipartitus, M. mohoity, and Barbatula toni) were investigated. Twelve morphological characters were measured in 542 individuals to perform the comparative analysis. Among these characters, only the caudal peduncle length (L_CP) revealed significant difference (P < 0.05) among these four species. The clustering based on morphological characters formed two clusters (P. dabryanus and M. anguillicaudatus; M. bipartitus and M. mohoity). A total of 186 COI fragments for the five loaches investigated were sequenced and analyzed. The results showed that interspecific K2P distance was much higher than intraspecific distance within the five species. Bayesian inference of phylogeny showed that individuals of these species were divided into five specific clades. Meanwhile, the COI fragments exhibited 22 character attributes for the differentiation of the five loach species based on character-based method. Our results suggested that DNA barcoding based on COI can be used as an efficient identifier of these five loach species; the combination of distance-based method, Bayesian inference and character-based approach provides higher resolution of identification at species level.     

The relative importance of geography and ecology in species diversification: evidence from a tropical marine intertidal snail (Nerita)

Aim In this study, I examined the relative contributions of geography and ecology to species diversification within the genus Nerita, a prominent clade of marine snails that is widely distributed across the tropics and intertidal habitats. Specifically, I tested whether geographical patterns of speciation correspond primarily to allopatric or sympatric models, and whether habitat transitions have played a major role in species diversification. Location Indo‐West Pacific, eastern Pacific, Atlantic, tropical marine intertidal. Methods I used a previously reconstructed molecular phylogeny of Nerita as a framework to assess the relative importance of geographical and ecological factors in species diversification. To evaluate whether recently diverged clades exhibit patterns consistent with allopatric or sympatric speciation, I mapped the geo‐graphical distribution of each species onto the species‐level phylogeny, and examined the relationship between range overlap and time since divergence using age–range correlation analyses. To determine the relative contribution of habitat transitions to divergence, I traced shifts in intertidal substrate affinity and vertical zonation across the phylogeny using parsimony, and implemented randomization tests to evaluate the resulting patterns of ecological change. Results Within the majority of Nerita clades examined, age–range correlation analysis yielded a low intercept and a positive slope, similar to that expected under allopatric speciation. Approximately 75% of sister species pairs have maintained allopatric distributions; whereas more distantly related sister taxa often exhibited complete or nearly complete geographical overlap. In contrast, only 19% of sister species occupy distinct habitats. For both substrate and zonation, habitat transitions failed to concentrate towards either the tips or the root of the phylogeny. Instead, habitat shifts have occurred throughout the history of Nerita, with a general transition from the lower and mid‐littoral towards the upper and supra‐littoral zones, and multiple independent shifts from hard (rock) to softer substrates (mangrove, mud and sand). Main conclusions Both geography and ecology appear to have influenced diversification in Nerita, but to different extents. Geography seems to play a principal role, with allopatric speciation driving the majority of Nerita divergences. Habitat transitions appear insignificant in shaping the early and recent history of speciation, and promoting successive diversification in Nerita; however, shifts may have been important for respective divergences (i.e. those that correspond to the transitions) and enhancing diversity throughout the clade.     

Foraging habits in a generalist predator: Sex and age influence habitat selection and resource use among bottlenose dolphins (Tursiops truncatus)

This study examines resource use (diet, habitat use, and trophic level) within and among demographic groups (males, females, and juveniles) of bottlenose dolphins (Tursiops truncatus). We analyzed the δ¹³C and δ¹⁵N values of 15 prey species constituting 84% of the species found in stomach contents. We used these data to establish a trophic enrichment factor (TEF) to inform dietary analysis using a Bayesian isotope mixing model. We document a TEF of 0‰ and 2.0‰ for δ¹³C and δ¹⁵N, respectively. The dietary results showed that all demographic groups relied heavily on low trophic level seagrass‐associated prey. Bayesian standard ellipse areas (SEA_b) were calculated to assess diversity in resource use. The SEA_b of females was nearly four times larger than that of males indicating varied resource use, likely a consequence of small home ranges and habitat specialization. Juveniles possessed an intermediate SEA_b, generally feeding at a lower trophic level compared to females, potentially an effect of natal philopatry and immature foraging skills. The small SEA_b of males reflects a high degree of specialization on seagrass associated prey. Patterns in resource use by the demographic groups are likely linked to differences in the relative importance of social and ecological factors.     

Odorella benthonica gen. & sp. nov. (Pleurocapsales,Cyanobacteria): an odor and prolific toxin producer isolated from a California aqueduct

Pleurocapsales are one of the least understood groups of cyanobacteria in terms of molecular systematics and biochemistry. Considering the high number of cryptic taxa within the Synechococcales and Oscillatoriales, it is likely that such taxa also occur in the Pleurocapsales. The new genus described in our research is the first known pleurocapsalean cryptic taxon. It produces off‐flavor and a large number of bioactive metabolites (n = 38) some of which can be toxic including four known microcystins. Using a polyphasic approach, we propose the establishment of the genus Odorella with the new species O. benthonica from material originally isolated from the California Aqueduct near Los Angeles.     

Getting a grip on the intertidal: flow microhabitat and substratum type determine the dislodgement of the crab Pachygrapsus crassipes (Randall) on rocky shores and in estuaries

Hydrodynamic forces can affect survival as well as limit the movement of motile benthic animals. An animal's danger of dislodgement depends on the hydrodynamic forces it experiences in its microhabitat relative to the force required to dislodge it (tenacity) from the substratum. We measured water flow and substratum characteristics in two different habitats of the shore crab Pachygrapsus crassipes: a wave-swept rocky shore and an intertidal mudflat. The maximum water velocities and accelerations in the microhabitats of the crabs at the wave-swept site were three times and two times greater, respectively, than at the mudflat site. In the laboratory, we measured the tenacity of crabs of various sizes on different substrata, and also measured their drag, lift and added-mass coefficients. Using these data, we calculated the flow conditions under which crabs would be overturned or sheared off the substratum in their two habitats. The net horizontal force (drag plus acceleration reaction) required to dislodge a crab on a rugose rock substratum was an order of magnitude greater than on smooth rock and two orders of magnitude greater than on mud. Our calculations indicate that, under non-storm conditions, crabs will not be dislodged from the substratum in either the mudflat or the wave-swept habitat when grasping the substratum with maximum tenacity. Moving crabs have lower tenacity and our calculations predict that hydrodynamic forces will restrict the mobility of large crabs more than that of small ones on smooth, but not on rugose rock.     

Molecular evidence for a species complex in the patagonian lizard Liolaemus bibronii and phylogeography of the closely related Liolaemus gracilis (Squamata: Liolaemini)

The lizard genus Liolaemus is endemic to temperate South America and includes 190 species. Liolaemus bibronii has a large geographic distribution and inhabits a great diversity of habitats, including the Monte, Steppe, and high Andean grassland environments. Liolaemus gracilis has a similar body size and shape to L. bibronii; the two are parapatrically distributed, and L. gracilis is also widely distributed. Here we use the mtDNA cytb sequence data of these two species to investigate lizard phylogeographic patterns in southern South America. L. bibronii is paraphyletic with respect to L. gracilis, Liolaemus ramirezae, Liolaemus robertmertensi and Liolaemus saxatilis; it is composed of many genetically different allopatric haploclades, some of which are reciprocally monophyletic. We also found evidence for introgression between L. bibronii and L. gracilis in the same area that introgression was hypothesized in the Liolaemus darwinii complex. We discuss the distribution of the major haploclades with inferences of their population histories, the concordance of these clades' distributions and histories with other lizard complexes studied with the same markers and methods, and taxonomic implications of these results.     

Laying date and clutch size of Great Tits(Parus major) in the Mediterranean region: a comparison of four habitat types

Summary Laying data and clutch size of Great Tits were studied in four different habitats in eastern Spain: two holm oak(Quercus ilex) forests, at 500 and 900–950 m a.s.l., a zeen oak(Quercus faginea) forest, at 900–1100 m a.s.l., a pine(Pinus sylvestris) forest, at 1000–1050 m a.s.l., and orange(Citrus aurantium) plantations, at 30 m a.s.l. All sites were placed at about the same latitude (39–41°N), and all were studied during the same years (1992–95). Our results show that (1) laying date did not differ between the natural habitats at the same altitude (range of the means of yearly means 4–8 May); (2) within the same habitat type (holm oak forest) laying date was earlier at low altitude (30 Aprilvs. 8 May); (3) laying date was earlier in the orange plantations (21 April) than in natural habitats; (4) among natural habitats at the same altitude, clutch size decreased from zeen oak (mean of yearly means 7.3 eggs) to holm oak (7.0 eggs) to pine forests (6.4 eggs), though only the difference between zeen oak and pine forests was significant; (5) within the same habitat type (holm oak forest), the clutch size tended to be larger at high altitude (7.0vs. 5.9 eggs); and (6) clutch size in orange plantations (7.7 eggs) did not differ significantly from that of the zeen oak forest, but was larger than in the holm oak and pine forests. We discuss the effect of the habitat type on laying date and clutch size of Great Tits.

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Legedatum und Gelegegröße der Kohlmeise(Parus major) in mediterranen Gebieten: Ein Vergleich zwischen vier verschiedenen Biotopen

Zusammenfassung Legedatum und Gelegegröße der Kohlmeise wurden in vier unterschiedlichen Biotopen in Ostspanien untersucht: zwei Steineichenwälder(Quercus ilex) in 500 m und 900–950 mNN, ein Bergeichenwald(Quercus faginea) in 900–1100 mNN, ein Kiefernwald(Pinus sylvestris) in 1000–1050 mNN und eine Orangenpflanzung(Citrus aurantium) in 30 mNN. Alle fünf Gebiete lagen auf etwa demselben Breitengrad (39–41°N) und wurden 1992–1995 parallel untersucht.(1) Auf gleicher Meereshöhe unterscheidet sich der Legebeginn nicht zwischen den verschiedenen Waldbiotopen (im Mittel 4.–8. Mai). (2) Innerhalb desselben Biotoptyps (Steineichenwald) war der Legebeginn auf niedrigerer Meereshöhe früher als in höheren Lagen (30. April vs. 8. Mai). (3) Im Orangenhain wurde früher mit der Eiablage begonnen (21. April) als in den Waldbiotopen. (4) Auf gleicher Meereshöhe nahm die Gelegegröße vom Bergeichenwald (Mittelwert 7.3 Eier) über die Steineichenwälder (7,0 Eier) zum Kiefernwald hin ab (6.4 Eier), jedoch ist nur der Unterschied zwischen Bergeichenwald und Kiefernwald signifikant. (5) Innerhalb der Steineichenwälder besteht die Tendenz zu größerer Gelegegröße in den höheren Lagen (7.0 vs. 5.9 Eier). (6) Im Orangenhain war die Gelegegröße mit durchschnittlich 7.7 Eier ähnlich der im Bergeichenwald, aber größer als in den Steineichen- und Kiefernwälder.

     

苍耳甾醇物质对菜青虫取食、血淋巴和中肠酶活性及中肠组织的影响

甾醇是植物体内的重要次生物质,具有多种生物活性。为探明植物甾醇类物质对害虫的作用机理,采用叶碟饲喂法进行取食处理后研究了苍耳Xanthium sibiricum中分离纯化的甾醇类组分（甾醇A和甾醇B）对4龄菜青虫Pieris rapae的取食、酶活性以及中肠组织的影响。结果表明： 苍耳甾醇类组分甾醇A和甾醇B能明显抑制菜青虫的取食,拒食中浓度AFC50分别为0.0229 和0.0147 mg/mL;同时,显著降低菜青虫中肠蛋白酶、淀粉酶和羧酸酯酶活性,其中,甾醇B的作用效果较强,处理后24 h和36 h, 对蛋白酶活性抑制率分别为23.74%和58.59%,对中肠羧酸酯酶的活性抑制率分别为49.01%和83.03%;降低血淋巴蛋白质含量,诱导菜青虫血淋巴羧酸酯酶活性的提高;破坏昆虫中肠上皮组织,微杆模糊不清呈消融状,杯状细胞的杯腔基部微绒毛消失。这些结果说明苍耳甾醇类物质对菜青虫的取食抑制可能与对中肠消化酶活性的抑制以及对中肠上皮组织的破坏有关,植物甾醇组分的不同配比影响其对昆虫的作用效果。