Linkage Analysis of the Multilocus Glucosephosphate Isomerase Isozyme System in Sunfish (Centrarchidae, Teleostii)

The purpose of the present investigation is to determine whether the two duplicated glucosephosphate isomerase (EC 5.3.1.9) loci Gpi-A and Gpi-B reside on the same chromosome in teleostean fishes. Interspecific sunfish hybrids were employed for the cross because of the different species-specific electrophoretic mobilities of the allelic isozymes at each GPI locus and because of their genomic compatibility. F1 sunfish hybrids, formed from a male warmouth (Lepomis gulosus) X female green sunfish (L. cyanellus) cross, were mated to form the F2 generation. The number of each of the nine different isozyme phenotypes, revealed by starch gel electrophoresis, was determined using 256 F2 individuals. The high frequency of recombinant phenotypes in the F2 generation indicated that the two GPI loci are not linked. An excess of F2 individuals heterozygous at both loci was observed and is interpreted as being caused by heterosis. The absence of linkage for the homologous loci encoding GPI subunits and for other multilocus isozyme systems is consistent with the postulate that the genomes of present-day vertebrates arose through one or more polyploidization events early in vertebrate evolution.     

Cross‐amplification of 10 new isolated polymorphic microsatellite loci for red mullet (Mullus barbatus) in striped red mullet (Mullus surmuletus)

Ten polymorphic dinucleotide microsatellite loci were isolated and characterized for the red mullet (Mullus barbatus). Allele variability was tested on both the red mullet and its congener the striped red mullet (Mullus surmuletus). Characterization of 30 individuals of both species from the western Mediterranean showed moderate to high allelic diversity ranging from two to 26 alleles per locus (mean 10.9). Three loci showed departures from Hardy–Weinberg proportions. No evidence of significant association between genotypes at pairs of loci was observed. These polymorphic loci could be suitable for population genetic assessments of both species.     

Multiple vitellogenin-derived yolk proteins in gray mullet (Mugil cephalus): disparate proteolytic patterns associated with ovarian follicle maturation

Disparate proteolytic patterns of yolk proteins, derived from three types of vitellogenin (VgA, VgB, and VgC), were observed in gray mullet. Immuno-biochemical analyses of extracts obtained from vitellogenic ovaries (VO) and ovulated eggs (OE) confirmed that a large proportion of VgA-derived lipovitellin (LvA) was degraded into free amino acids (FAAs) during ovarian follicle maturation. The maturation-associated alteration of VgB-derived Lv (LvB) involved only limited proteolysis; the heavy and light LvB chains were dissociated into at least three and one polypeptide fragments, respectively. The native mass of VgC-derived Lv (LvC) exhibited little difference between VO and OE, although it was apparent that the LvC was 'nicked' during maturation, resulting in the appearance of several bands in OE. Similar analyses confirmed that VgA-derived beta'-component (beta'-cA) and VgB-derived beta'-c (beta'-cB) decreased during maturation in both quantity and native mass, while phosvitin derived from either VgA (PvA) or VgB (PvB) appeared to be degraded into FAAs. The pattern of maturation-associated proteolysis of mullet yolk proteins is similar to that reported for other marine teleosts spawning pelagic eggs. However, the depository ratio of the three distinct types of Lv in the mullet VO appeared to be different from that estimated for another marine pelagophil, the barfin flounder. These results support a recent paradigm regarding the significance of Vg multiplicity upon successive physiological events in this group of fishes including the hydration of maturing oocytes, the acquisition of proper egg buoyancy, and the generation of requisite nutrient stocks for each stage of embryogenesis and larval development.     

Bacteriological aspects of fin erosion in mullet exposed to crude oil

An investigation was undertaken to determine the effect of two types of crude oil on mullet ( Mugil cephalus ). The data presented in this report establishes a relationship between the exposure of the mullet to crude oils and the outbreak of fin rot. The data also indicates that the diversity of the microflora on the exterior of the mullet was reduced from 5-6 to 1-2 colonial types, one of which was consistently a species of the genus Vibrio.     

First Microsatellite Loci of Red Mullet (<Emphasis Type="Italic">Mullus barbatus</Emphasis>) and Their Application to Genetic Structure Analysis of Adriatic Shared Stock

In order to study the genetic structure of the Adriatic shared stock of red mullet (Mullus barbatus), we developed a set of dinucleotide microsatellite markers. A dinucleotide-enriched genomic library was obtained, and 6 polymorphic dinucleotide loci were successfully optimized. The markers showed high expected heterozygosity (from 0.68 to 0.92) and allele number (from 12 to 33); thus they appear to be suitable for detecting genetic differences in the population of red mullet. Four Adriatic samples were subsequently analyzed for microsatellite variation, and the results showed subtle but statistically significant genetic differentiation, indicating that the Adriatic red mullet may group into local, genetically isolated populations. No correlation between geographic distance and genetic differentiation was observed. In addition, the evidence of recent bottlenecks in the Adriatic samples indicates that the observed population subdivision might reflect random local allelic variations, generated by reproductive success, survival rates, or fishing pressure.     

Perceived wintering latitude determines timing of song output in a migratory bird

Migratory bird populations frequently consist of individuals that overwinter variable distances from the breeding site. Seasonal changes in photoperiod, which varies with latitude, underlie seasonal changes in singing frequency in birds. Therefore, migratory populations that consist of individuals that overwinter at different latitudes with large overwintering ranges could experience within‐population variation in seasonal production of song. To test the influence of overwintering latitude on intrapopulation variance in song production in the spring, we subjected two groups of Eastern Song Sparrows (Melospiza melodia melodia) from the same partially migratory breeding population to different photoperiodic schedules associated with a 1,300‐km difference in overwintering location. One group remained on the natural photoperiodic schedule of the breeding site (resident group) while the other group experienced a nonbreeding photoperiod that mimicked a southern migration in the fall followed by a northern migration back to the breeding site in the spring (migratory group). We compared song output between the two groups in three different stages (nonbreeding, prebreeding, and breeding). Little singing occurred during nonbreeding stage sample dates (20 November, 6 December) for the resident group, and no singing occurred for the migrant group. During the prebreeding stage (27 January, 7 February), significantly more singing occurred in the resident group than in the migrant group. During the breeding stage (21 March, 4 April), after a simulated migration for the migrants, song output was similar in both groups. These results suggest that within‐population variation in wintering latitude may contribute to variation in seasonal changes in singing behavior, which may covary with readiness to breed. Studies utilizing confirmed migrants and residents, rather than merely simulated migrants and residents, are also needed to better understand these processes.     

Demographic and spatial responses of resident bird populations to the arrival of migratory birds within an urban environment

Long-distance migration allows many bird species to overcome the severe climatic changes that occur in seasonal environments. Migration is highly demanding, and given its cyclical nature, we currently know that it has substantial effects on the population parameters of migratory birds during both breeding and wintering seasons. However, the potential effects of the presence of migratory birds in their wintering grounds on populations of resident birds have remain largely unexplored. Here, we propose the hypothesis that migratory birds negatively affect the habitat occupancy and population abundance of resident birds because of the arrival of numerous individuals during the most limiting months of the year. Here, we studied different species of migratory and resident birds that coexist during winter in an urban ecological reserve located within Mexico City. We used single-species multiseason occupancy models, two-species occupancy models, and distance sampling techniques to evaluate changes in occupancy and population density of resident bird species during three consecutive winters. We found an aggregation pattern between two resident species (Psaltriparus minimus and Thryomanes bewickii) with three migratory warblers (Cardellina pusilla, Setophaga coronata and Setophaga townsendi). Thus, our results provide evidence of the formation of mixed-species flocks in our study area. We also conclude that resident birds experience different demographic and behavioral processes during winter that not necessarily result from interspecific interactions with migratory birds.     

Genetic divergence at the NAD +-dependent malic enzyme locus in Atlantic salmon from Europe and North America

The distribution of genetic variation for NAD+ malic enzyme ( ME* ) polymorphism in the Atlantic salmon was assessed in both anadromous and resident populations. The analysis revealed a major allelic divergence between North America and Europe. The *80 variant occurred in 39 of 40 North American samples, ranging in frequency from 0 to 0.515. In contrast, it was detected in only three fish from two of the 35 European locations analysed. Eleven fish from two rivers in north-west France had the * 110 variant. While heterogeneity among North American populations was significant, no regional differentiation was apparent and anadromous and resident salmon were not found to differ.     

Temporal dynamics of migration‐linked genetic variation are driven by streamflows and riverscape permeability

Landscape permeability is often explored spatially, but may also vary temporally. Landscape permeability, including partial barriers, influences migratory animals that move across the landscape. Partial barriers are common in rivers where barrier passage varies with streamflow. We explore the influence of partial barriers on the spatial and temporal distribution of migration‐linked genotypes of Oncorhynchus mykiss, a salmonid fish with co‐occurring resident and migratory forms, in tributaries to the South Fork Eel River, California, USA, Elder and Fox Creeks. We genotyped >4,000 individuals using RAD‐capture and classified individuals as resident, heterozygous or migratory genotypes using life history‐associated loci. Across four years of study (2014–2017), the permeability of partial barriers varied across dry and wet years. In Elder Creek, the largest waterfall was passable for adults migrating up‐river 4–39 days each year. In this stream, the overall spatial pattern, with fewer migratory genotypes above the waterfall, remained true across dry and wet years (67%–76% of migratory alleles were downstream of the waterfall). We also observed a strong relationship between distance upstream and proportion of migratory alleles. In Fox Creek, the primary barrier is at the mouth, and we found that the migratory allele frequency varied with the annual timing of high flow events. In years when rain events occurred during the peak breeding season, migratory allele frequency was high (60%–68%), but otherwise it was low (30% in two years). We highlight that partial barriers and landscape permeability can be temporally dynamic, and this effect can be observed through changing genotype frequencies in migratory animals.     

Partial migration: growth varies between resident and migratory fish

Partial migration occurs in many taxa and ecosystems and may confer survival benefits. Here, we use otolith chemistry data to determine whether fish from a large estuarine system were resident or migratory, and then examine whether contingents display differences in modelled growth based on changes in width of otolith growth increments. Sixty-three per cent of fish were resident based on Ba : Ca of otoliths, with the remainder categorized as migratory, with both contingents distributed across most age/size classes and both sexes, suggesting population-level bet hedging. Migrant fish were in slightly better condition than resident fish based on Fulton''s K condition index. Migration type (resident versus migratory) was 56 times more likely to explain variation in growth than a model just incorporating year- and age-related growth trends. While average growth only varied slightly between resident and migratory fish, year-to-year variation was significant. Such dynamism in growth rates likely drives persistence of both life-history types. The complex relationships in growth between contingents suggest that management of species exhibiting partial migration is challenging, especially in a world subject to a changing climate.     

On fitness and partial migration in a large herbivore – migratory moose have higher reproductive performance than residents

Partially migratory populations comprise both resident and migratory individuals. These tactics may coexist if their demographic contribution to future generations (i.e. fitness) are equal or vary temporally with environmental conditions, or if individuals switch between being migrant and resident. Alternatively, the choice of movement tactic can be based on individual attributes such as age, competitive ability or personality. In the latter cases, the two tactics are not expected to have similar average fitness. In this study, we examined the effect of movement tactic on reproductive performance and survival of 82 GPS‐marked female moose and their offspring in a partial migratory population in central Norway. The results indicated higher growth in the migrating part of the population because migrating females produced more twins than resident females. We found no differences in pregnancy rates or survival of adults or their offspring, indicating a net fitness benefit of being migrant. We found the average shoulder height of residents to be slightly lower than of migrants, but doubt that this affected their migration ability. A more likely explanation is that migratory females are both more fecund and grow bigger because of better conditions in their summer ranges. This may be a temporal phenomenon if the fitness differences between migratory and resident moose vary according to environmental fluctuations.     

Structural and functional differentiation of two clinally distributed glucosephosphate isomerase allelic isozymes from the teleost Fundulus heteroclitus

The teleost Fundulus heteroclitus (L.) possesses two loci, Gpi-A and Gpi-B, for the glycolytic enzyme, glucose-phosphate isomerase (GPI; D- glucose-6-phosphate ketol-isomerase; E.C. 5.3.1.9). The Gpi-B locus is polymorphic in Fundulus, with two common alleles, Gpi-Bb and Gpi-Bc, distributed in a clinal manner in populations along the east coast of North America. Since this clinal distribution is strongly correlated with a temperature gradient, we asked whether the GPI-B2 allozymes were functionally adapted to the thermal environment in which a given phenotype predominated. The two major GPI-B2 allozymes were purified to homogeneity and were characterized as to molecular weight, isoelectric pH, thermal denaturation, and kinetic parameters. Both GPI-Bb2 and GPI- Bc2 allozymes have molecular masses of 110 kD, and they have isoelectric pHs of 6.4 and 6.6, respectively. The GPI-Bb2 allozyme was more stable to thermal denaturation than was the GPI-Bc2 enzyme. Kinetic properties of the allelic isozymes were investigated both as a function of pH and as a function of temperature. At 25 degrees C, over the pH range considered, there were no significant differences between allozymes, either in Km for fructose-6-phosphate or in Ki for 6- phosphogluconate, but apparent Vmax values differed between pH 7.5 and pH 8.5. All steady-state kinetic parameters showed strong temperature dependence, but the allozymes differed only in the Ki for 6- phosphogluconate at temperatures greater than 30 degrees C. On the basis of the observed structural and functional differences alluded to above, the hypothesis that the major allelic isozymes of the Gpi-B locus were functionally equivalent was rejected. However, it is not yet known whether these structural and functional differences have any significance at higher levels of biological organization.      

Assessing the effects of habitat characteristics and co-occurrence with closely related species on occupancy of resident ‘wood-warblers’ in a Neotropical working landscape

The simultaneous effects of habitat traits and interspecific interactions determine the occurrence and habitat use of wildlife populations. However, little research has been devoted to examining spatial co-occurrence among closely related species while considering the effect of habitat variation and imperfect detectability of species in the field. In this study, we focused on migratory and resident 'wood-warblers' that coexist during the winter in a Neotropical working landscape in southern Mexico to understand if habitat occupancy of resident wood-warblers is influenced by habitat characteristics and by the presence of other species of resident and migratory wood-warblers. For this purpose, we implemented two-species occupancy models, which account for the imperfect detectability of these birds in the field. Our results revealed that habitat occupancy of resident wood-warblers was positively influenced by the presence of other closely related species (both migratory and resident). These positive relationships may be explained by the fact that different species of wood-warblers frequently participate in mixed-species flocks. However, these patterns of species co-occurrence were more evident among resident species than between migratory and resident species, which may be explained by micro-habitat segregation and differences in behaviours between resident and migratory wood-warblers. We also found that some habitat characteristics may mediate the observed patterns of species co-occurrence. Specifically, sites with larger trees were associated with the co-occurrence of some species of resident wood-warblers. In addition, we discuss the possibility that species co-occurrence might be the result of shared preferences for environmental factors that we did not consider. Our study highlights the importance of the interplay between species co-occurrence and habitat traits in determining the presence and habitat use of resident birds in Neotropical working landscapes.     

A comparison of sex ratio,birth periods and calf survival among Serengeti wildebeest sub‐populations,Tanzania

Although adaptation and environmental conditions can easily predict demographic variation in most savannah ungulates, no study on demographic consequences arising from natural and anthropogenic factors among Serengeti wildebeest (Connochaetes taurinus) sub‐populations in Tanzania has been carried out. Here, I report estimates of annual sex ratio, calf and yearling survival rate and birth seasonality between resident and migratory sub‐populations to explore demographic patterns arising from the different age and sex structure. The results indicate significantly higher female‐biased sex ratios in the resident and almost even sex ratios among individual migrants. The calf recruitment estimated as mother: offspring ratios indicate a more synchronous birth in the migrant than the resident sub‐population. Also, birth seasonality in the migratory sub‐population coincided with seasonal variability of rainfall and the timing of the birth peak was more variable in the migrants than the resident sub‐population. The migratory sub‐population had a higher annual proportional mean calf survival estimate (0.84) than that of the residents (0.44) probably due to higher mortality resulting from predation in the western corridor. However, the proportion of yearling survival estimates was much lower (0.31) in the migrants and relatively higher (0.39) in the residents. Different demographic outcomes resulting from environment, predation, movements and ecological factors including resource competition have conservation implications for the two sub‐populations.     

Detecting shifts of transmission areas in avian blood parasites: a phylogenetic approach

We investigated the degree of geographical shifts of transmission areas of vector-borne avian blood parasites (Plasmodium, Haemoproteus and Leucocytozoon) over ecological and evolutionary timescales. Of 259 different parasite lineages obtained from 5886 screened birds sampled in Europe and Africa, only two lineages were confirmed to have current transmission in resident bird species in both geographical areas. We used a phylogenetic approach to show that parasites belonging to the genera Haemoproteus and Leucocytozoon rarely change transmission area and that these parasites are restricted to one resident bird fauna over a long evolutionary time span and are not freely spread between the continents with the help of migratory birds. Lineages of the genus Plasmodium seem more freely spread between the continents. We suggest that such a reduced transmission barrier of Plasmodium parasites is caused by their higher tendency to infect migratory bird species, which might facilitate shifting of transmission area. Although vector-borne parasites of these genera apparently can shift between a tropical and a temperate transmission area and these areas are linked with an immense amount of annual bird migration, our data suggest that novel introductions of these parasites into resident bird faunas are rather rare evolutionary events.     

Host immune defence and migration in birds

Migratory birds are exposed to at least two different parasite faunas during their annual cycle, while resident birds only experience a single parasite fauna. Migratory birds should therefore have evolved mechanisms to control or reduce the negative impact of infections from a more diverse parasite fauna. In a comparison of pairs of closely related species of birds that differ with respect to whether they are migratory or residents, the size of two immune defence organs (the bursa of Fabricius and the spleen) was consistently larger in the migratory species. Since the bursa is only found in juvenile, sexually immature birds, we conclude that immune defence adaptations to the impact of a more diverse parasite fauna in migrants already exist before the start of the first migration. Interspecific differences in investment in immune defence between migratory and resident birds have implications for our understanding of complex host–parasite interactions, the acquisition of new hosts by parasites, and the susceptibility of migratory birds to environmental perturbations.     

tfoX (sxy)-dependent transformation of Aggregatibacter (Actinobacillus) actinomycetemcomitans

tfoX (sxy) is a regulatory gene needed to turn on competence genes. Aggregatibacter (Actinobacillus) actinomycetemcomitans has a tfoX gene that is important for transformation. We cloned this gene on an IncQ plasmid downstream of the inducible tac promoter. When this plasmid was resident in cells of A. actinomycetemcomitans and tfoX was induced, the cells became competent for transformation. Several strains of A. actinomycetemcomitans, including different serotypes, as well as rough (adherent) and isogenic smooth (nonadherent) forms were tested. Only our two serotype f strains failed to be transformed. With the other strains, we could easily get transformants with extrachromosomal plasmid DNA when closed circular, replicative plasmid carrying an uptake signal sequence (USS) was used. When a replicative plasmid carrying a USS and cloned DNA from the chromosome of A. actinomycetemcomitans was linearized by digestion with a restriction endonuclease or when genomic DNA was used directly, the outcome was allelic exchange. To facilitate allelic exchange, we constructed a suicide plasmid (pMB78) that does not replicate in A. actinomycetemcomitans and carries a region with two inverted copies of a USS. This vector gave allelic exchange in the presence of cloned and induced tfoX easily and without digestion. Using transposon insertions in cloned katA DNA, we found that as little as 78 bp of homology at one of the ends was sufficient for that end to participate in allelic exchange. The cloning and induction of tfoX makes it possible to transform nearly any strain of A. actinomycetemcomitans, and allelic exchange has proven to be important for site-directed mutagenesis.     

Rufous‐capped Warblers Basileuterus rufifrons show seasonal,temporal and annual variation in song use

In the majority of songbird species, males have repertoires of multiple song types used for mate attraction and territory defence. The wood‐warblers (family Parulidae) are a diverse family of songbirds in which males of many migratory species use different song types or patterns of song delivery (known as ‘singing modes’) depending on context. The vocal behaviour of most tropical resident warblers remains undescribed, although these species differ ecologically and behaviourally from migratory species, and may therefore differ in their vocal behaviour. We test whether male Rufous‐capped Warblers Basileuterus rufifrons use distinct singing modes by examining song structure and context‐dependent variation in their songs. We recorded multiple song bouts from 50 male Warblers in a Costa Rican population over 3 years to describe seasonal, diel and annual variation in song structure and vocal behaviour. We found that Rufous‐capped Warbler songs are complex, with many syllable types shared both within and between males’ repertoires. Males varied their song output depending on context: they sang long songs at a high rate at dawn and during the breeding season, and shortened songs in the presence of a vocalizing female mate. Unlike many migratory species, Rufous‐capped Warblers do not appear to have different singing modes; they did not change the song variants used or the pattern of song delivery according to time of day, season or female vocal activity. Our research provides the first detailed vocal analysis of any Basileuterus warbler species, and enhances our understanding of the evolution of repertoire specialization in tropical resident songbirds.     

Isolation and characterization of visceral excitatory neuropeptides from striped mullet (Mugil cephalus L.) brain

From a brain extract of the catadromous fish, striped mullet (Mugil cephalus), two visceral excitatory neuropeptides (Mvp-1 and Mvp-2) were isolated by means of reversed phase chromatography together with bioassay on fish hindgut. The primary structure of Mvp-1 was elucidated to be SGPAGVLamide by ESI-Q-TOF mass spectrometry. The threshold concentration of Mvp-1 that changes spontaneous contraction of mullet hindgut was between 10(-9) and 10(-8) M. In addition, Mvp-1 was found to exert excitatory activities on some other smooth muscle segments (oviduct and esophagus) of mullet but it did not show any effect on body wall muscle strips. Therefore, the present study suggests that Mvp-1 and Mvp-2 peptides act as factors that control visceral contractions of mullet gastrointestinal tract.     

Phenological sensitivity to climate change is higher in resident than in migrant bird populations among European cavity breeders

Many organisms adjust their reproductive phenology in response to climate change, but phenological sensitivity to temperature may vary between species. For example, resident and migratory birds have vastly different annual cycles, which can cause differential temperature sensitivity at the breeding grounds, and may affect competitive dynamics. Currently, however, adjustment to climate change in resident and migratory birds have been studied separately or at relatively small geographical scales with varying time series durations and methodologies. Here, we studied differential effects of temperature on resident and migratory birds using the mean egg laying initiation dates from 10 European nest box schemes between 1991 and 2015 that had data on at least one resident tit species and at least one migratory flycatcher species. We found that both tits and flycatchers advanced laying in response to spring warming, but resident tit populations advanced more strongly in relation to temperature increases than migratory flycatchers. These different temperature responses have already led to a divergence in laying dates between tits and flycatchers of on average 0.94 days per decade over the current study period. Interestingly, this divergence was stronger at lower latitudes where the interval between tit and flycatcher phenology is smaller and winter conditions can be considered more favorable for resident birds. This could indicate that phenological adjustment to climate change by flycatchers is increasingly hampered by competition with resident species. Indeed, we found that tit laying date had an additional effect on flycatcher laying date after controlling for temperature, and this effect was strongest in areas with the shortest interval between both species groups. Combined, our results suggest that the differential effect of climate change on species groups with overlapping breeding ecology affects the phenological interval between them, potentially affecting interspecific interactions.