Genetic and morphological studies of Nothobranchius (Cyprinodontiformes) from Malawi with description of Nothobranchius wattersi sp. nov.

Molecular and morphological data were used to explore evolutionary differentiation among populations of Nothobranchius in the Lake Malawi–upper Shire River and the Lakes Chilwa–Chiuta drainage systems in Malawi. The aim of the study was to test the hypothesis that Nothobranchius of the Malawi–Shire system constitute a separate evolutionary group from Nothobranchius kirki. Mitochondrial and nuclear sequence data show a strongly supported phylogenetic split into two monophyletic groups separating the Lake Malawi basin fish from N. kirki. Unlike N. kirki, Lake Malawi–Shire fish do not deviate from neutrality and express an excess of rare haplotypes and mutations in terminal branches, characteristic of recently expanded populations. Further, the two groups significantly differ in morphology. Two body characters (dorsal‐fin base length and pre‐pelvic–pre‐anal distance) are significantly different between the two species in both sexes. Several other characters are significantly different in either male or female comparisons with respect to both standard and head lengths, and robust morphological differentiation is detected by multivariate analysis. The two groups are readily distinguished on the basis of male colouration, especially in scale centres and the caudal fin. On the basis of this differentiation at the molecular and morphological levels, in addition to colouration, the Lake Malawi–Shire fish are hereby formally recognized as constituting a new species, Nothobranchius wattersi. This distinction is in agreement with the geomorphologic and recent climatic history in the region.     

Causes of mortality in the endemic Tilapia of Lake Chilwa (Malawi)

Summary During 1966 and 1967 extensive mortalities of Tilapia shirana chilwae took place in Lake Chilwa, Malawi. By collating field observations and physico-chemical data from the lake with experimental evidence from the laboratory, it has been possible to predict what the most likely causes of mortality of Tilapia might have been.High winds caused bottom erosion in October 1966, and the released mud deoxygenated the water. Total deoxygenation of the water results if the level of suspended matter is raised to 12,860 ppm. Tilapia are able to survive levels of oxygen as low as 0.6 mg/l, but become distressed below this figure.The median lethal alkalinity for T. s. chilwae in artificial lake water is 61.6 meq/l. This concentration was reached in the lake in December 1966, and probably caused further mortalities. T. s. chilwae is remarkably tolerant of high temperature, and this factor alone is unlikely to have caused natural mortalities. Elevated temperatures can cause decreased resistance to low oxygen and high ionic concentrations, however, and the dense blooms of Anaebena may have caused sufficient deoxygenation of the water at night to cause further mortalities. T. s. chilwae is more resistant to high alkalinity and high temperature than T. melanopleura. This undoubtedly reflects the long history of stress in the endemic Tilapia. No doubt the heightened resistance of T. s. chilwae has enabled it to successfully populate L. Chilwa, despite its characteristic instability.

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Zusammenfassung Betrachtliche Mortalitäten des Fisches Tilapia shirana chilwae erfolgte im Chilwa See (Malawi) in 1966 und 1967. Bei der Zusammenfügung von den Beobachtungen am See und den physischchemischen Daten des Sees mit experimentellen Daten aus dem Laboratorium, war es moglich die wahrscheinlichen Ursachen des Mortalitäts festzustellen.Starke Winde verursachten die Erosion des Bodens im Oktober 1966 und der aufgelöste Schlamm entfernte die Sauerstoffe des Wassers. Totale Entfernung des Sauerstoffes folgte als tas Niveau des aufgelösten Materials einen Wert von 12,800 t.p.m. erreichte. Obgleich Tilapia einen so niedrigen Sauerstoffniveau von 0.6 mg/l überleben können fühlen she sich unheimlich unter diesem Wert.Die mediane letale Alkalinitat für T. s. chilwae im künstlichen Seewasser ist 61.6 meq/l. Diese Konzentration wurde im Dezember 1966 im See erreicht und verursachte wahrscheinlich eine weitere Mortalität. T. s. chilwae ist auffällig tolerant hohen Temperaturen gegen-über und es ist unwahrscheinlich dasz these Tatsache allein natürliche Mortalitäten verursacht hat. Dennoch können erhdhte Temperaturen eine abnehmende Resistenz gegen niedrigen Sauerstoff und hohe Ionen-Konzentration verursachen und die dicke Bliite von Anaebena könnte eine genügende Desoxygenation des Wassers in der Nacht veranlasst haben um weitere Mortalitaten zu verursachen. T. s. chilwae zeigt stärkere Resistenz gegenüber hoher Alkalinität und hoher Temperaturen als T. melanopleura. Dieses reflektiert, ohne Zweifel die lange Geschichte von Stress in die endemische Tilapia. Die erhöhte Resistenz des T. s. chilwae, dasz es für these Fische möglich war den Chilwa See zu bevölkern, trotz ihrer charakteristischen Instabilität.

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University of Malawi     

THE FOOD OF SOME DUCK SPECIES OCCURRING AT LAKE CHILWA,MALAWI

Schulten, G. G. M. 1974. The food of some duck species occurring at Lake Chilwa, Malawi. Ostrich 45: 224–226.

Gullet and stomach contents of 23 Southern Pochards, 17 Fulvous Treeducks, 14 White-faced Treeducks, 15 Hottentot Teals, 4 Redbilled Teals, 1 Whitebacked Dack and 1 Pigmy Goose were analysed. These birds were shot in the Lake Chilwa area (Malawi, C. Africa) between 1969–1972. Most of the food components were identified and their occurrence is presented in a table. The results are discussed.     

Abundance and distribution of fish in the lower Mnembo River, Malawi-Mozambique

Endorheic Lake Chilwa is one of the most productive lakes for fisheries in Africa. High human population densities and agricultural practices in the Chilwa catchment have been linked to observed declines of the annual fish catch in the lake. The Mnembo River is a major inflow into Lake Chilwa which has received little scientific study to date. In 2003/2004, fish diversity, abundance and distribution were monitored monthly at three sites in the Mnembo River. Twelve fish species were collected, a subset of the fourteen fish species present in Lake Chilwa. Although Barbus spp. were the most abundant species sampled in the Mnembo, total biomass was higher for Labeo cylindricus (6709 and 10,434 g, respectively). Total biomass of Pareutropius longifilis (1741 g) and Brycinus imberi (1174 g) were also high in the river. Catches of Barbus and Labeo were highly correlated (r = 0.763). Compared with other inflows into Lake Chilwa (Likangala and Domasi), the Mnembo River appears to be less adversely influenced by human populations, with high abundance of Barbus and strong presence of the riverine species Labeo, Pareutropius and Brycinus.     

Zooplankton in a quasi-stable phase in an endorheic lake (Lake Chilwa,Malawi)

Lake Chilwa, a fairly large, turbid, brackish and very shallow endorheic lake in Malawi, Central Africa, dried up completely in 1968 and filled up again in the following wet season. Compared with the zooplankton in the drying, filling and post-filling phases (1966–1971), differences in their composition, density, distribution and reproduction were found in 1975–1976, a year when the lake was 1–2 metres deeper. This situation is discussed in relation to environmental factors, fish predation and the supply of detritus from the surrounding Typha swamp, to illustrate the relative stability of the zooplankton populations in periods between lake recessions.University of Malawi     

The role of the littoral zone in the functioning of a shallow tropical lake ecosystem

Lake Chilwa (Malawi) is a shallow tropical lake surrounded by an extensive zone of littoral swamp mainly composed of Typha domingensis. The data produced show that the physical and chemical conditions are different and more varied in the littoral region when compared with the open lake. Primary production in the littoral is confined almost entirely to Typha domingensis, which also acts as a nutrient pump by absorbing nutrients from the soils and eventually releasing them into the lake waters. Long term and short term exchanges in organic and inorganic materials between the littoral swamps and the open lake are described. These, as well as available evidence on the distribution, reproductive habits and diet of certain faunal groups, indicate that the littoral region of Lake Chilwa has an important regulating influence on the functioning of the whole lake.     

Ten polymorphic microsatellite loci for the endangered Buena Vista Lake shrew (Sorex ornatus relictus)

The ornate shrew (Sorex ornatus) is restricted to the vanishing wetlands of California, USA and Baja California, Mexico. Several subspecies of ornate shrews are considered ‘mammal species of special concern’ in California by the Department of Fish and Game, and one (Sorex ornatus relictus) has recently been listed as endangered. Populations of shrews around Buena Vista Lake have been diminished or extirpated due to habitat deterioration and human development. In order to study the patterns of genetic variation in isolated populations of Buena Vista Lake shrews, we developed 10 polymorphic microsatellite loci. There were 6–27 alleles per locus, and the loci had heterozygosity values that ranged from 20 to 80%. In addition, we screened 20 different populations of S. ornatus, eight species within two subfamilies of shrews (Soricinae and Crocidurinae), as well as in a mole (Talpidae, Neurotrichus gibbsii), to determine if these loci could be informative in other species as well.     

Elevated mtDNA diversity in introduced populations of Cynotilapia afra (Günther 1894) in Lake Malawi National Park is evidence for multiple source populations and hybridization

Genetic variation in many invasive species shows little or no signs of a founder event, suggesting that high genetic diversity may facilitate establishment success. The rocky‐shore, plankton‐feeding cichlid fish Cynotilapia afra is endemic to Lake Malawi, but naturally absent from many suitable sites. In the 1960s, this species was introduced to the southern areas of the lake, presumably as a result of the aquarium fish trade. It has now become established on a number of rocky areas within the Lake Malawi National Park. Here, we analysed DNA sequence variation in the mitochondrial control region of six native and four introduced populations of C. afra, and three populations of the closely‐related and hybridizing Pseudotropheus zebra. In contrast to previous studies of Lake Malawi rock dwelling cichlids, network analyses suggested that native populations of C. afra showed high levels of lineage sorting in mtDNA. Introduced populations showed higher sequence and haplotype diversity than their native counterparts. Our analyses suggested that the elevated gene diversity was largely attributed to the fact that the introduced C. afra populations were derived from several genetically distinct and geographically separate populations, and to a lesser extent because of introgressive hybridization with native P. zebra. The establishment and spread of C. afra may be partly because of its ability to occupy a vacant ecological niche, but it may also have been facilitated by its enhanced genetic diversity.     

Evolutionary relationships of the Lake Malawi Oreochromis species: evidence from allozymes

Three tilapiine species belonging to the endemic Lake Malawi species flock known as' chambo' Oreochromis ( Nyasalapia ) karongae , O. (N.) lidole and O. (N.) squamipinnis and the species Oreochromis (Oreochromis) shiranus were collected from the wild. These four species were analysed at 43 enzyme loci using starch gel electrophoresis. No significant deviations from Hardy-Weinberg equilibrium were observed after correcting for multiple simultaneous testing. The expected heterozygosity ( H _e) was lowest in O. (O.) shiranus ( H _e=0.082). The three chambo species had higher levels: H _e=0.103 to H _e=0.116. The three chambo species as a whole were polymorphic (99%) at 16 different loci sharing 10 of these in common. No fixed differences between the three chambo species were observed but highly significant allele frequency differences existed between all the chambo species. The F _ST calculated for the three chambo species was 0051, closer to intra-specific than to inter-specific levels. O. (O.) shiranus could be clearly separated from all chambo species at five fixed loci. Comparison of the allozyme data from these species with five species from the same sub genera support the hypothesis that the chambo form a monophyletic group and that O. (N.) macrochir or a related species represents the sister taxon.     

RAPD variation in wild populations of four species of the genus Hordeum (Poaceae)

 The genetic variation of 102 natural populations of wild barley growing in Spain was assessed using RAPDs (random amplified polymorphic DNA). The plant material included the annual species H. marinum subsp. marinum (22 populations) and subsp. gussoneanum (14), H. murinum subsp. murinum (7) and subsp. leporinum (35), and the perennial species H. bulbosum (17) and H. secalinum (7). Ten of the tested 64 arbitrary 10-mer primers amplified polymorphic DNA in all taxonomic units. Analyses was performed within and between populations, species and subspecies. The primers gave a total of 250 RAPD products. The level of polymorphism varied between taxonomic units depending on the primers employed and the plant reproductive system. In general, the most variable were the allogamous species H. secalinum and H. bulbosum and the autogamous H. marinum subsp. marinum. Among the amplified bands, 69 (27%) were shared by at least two different taxonomic units. The remaining bands were specific. The results demonstrate differences in the degree of similarity between taxonomic units. Jaccard’s similarity coefficients for interval measure within and between populations were used to produce a cluster diagram using the unweighted pair-group method (UPGMA). The different populations of the species and subspecies of Hordeum fell into three groups. The first group contained the populations belonging to both subspecies of H. marinum, plus those of H. secalinum. The populations of H. marinum subsp. gussoneanum were very closely associated. Those of H. marinum subsp. marinum were grouped in a broad cluster. The second group, occupying the innermost position of the tree, was very closely associated with the populations of both subspecies of H. murinum. The third branch segregated H. bulbosum. A series of RAPD markers were investigated by cleaving the amplified products of the same size with restriction endonucleases that recognize targets of 4- or 6-bp. The production of equivalent fragments following cleavage by the same enzyme would seem to demonstrate their homology in samples from different individuals, populations or taxonomic units. Received: 18 May 1997 / Accepted: 22 August 1997     

Genetic characterization of common carp (Cyprinus carpio) populations from Greece using mitochondrial DNA sequences

Wild common carp from two lakes and two rivers in Greece were genetically characterized with sequencing analysis of two mitochondrial DNA segments: cytochrome b (1119 bp) and D-loop (646 bp). A total of 9 variable singleton sites and 7 unique haplotypes were detected. A common haplotype was found in three out of the four populations examined, which seems to be the ancestral one and represents the European origin of common carp from Greece. This haplotype could be also justified by the introductions reported with individuals belonging to the Central European race, into many natural habitats in Greece. Limited genetic variation — in Evros and Aliakmonas populations — could be due to bottleneck effects and small effective population sizes, whereas the different haplotypes found in Lake Volvi could represent different common carp stocks. Values of sequence divergence among Greek haplotypes ranged from 0.0006 to 0.0023. The Neighbour-Joining (NJ) phylogenetic tree constructed based on the combined sequences, reveals that the populations of common carp from Greece belong to the European group of populations — which is highly divergent from the South East-Asia cluster — and to the subspecies Cyprinus carpio carpio.     

PATTERNS OF FLORAL NECTAR-SUGAR COMPOSITION OF IPOMOPSIS LONGIFLORA (POLEMONIACEAE) NEAR THE CONTACT ZONE OF ITS SUBSPECIES LONGIFLORA AND AUSTRALIS

The nectar-sugar compositions of 338 individuals from 27 populations in two subspecies of Ipomopsis longiflora from Arizona, New Mexico, Texas, and Chihuahua were determined by high-performance liquid chromatography. Mean sucrose compositions of populations ranged from 73.2% to 91.9%, and the standard deviations ranged from 1.9% to 9.1%. Pairs and aggregates of populations were statistically compared. The nectar-sugar compositions of the two subspecies were significantly different as aggregates. Subspecies australis averaged 9.5% fructose, 7.9% glucose, and 82.6% sucrose while subspecies longiflora averaged 7.8% fructose, 5.7% glucose, and 86.5% sucrose. Each subspecies was found to have two groups of populations. Groups of higher sucrose populations were found along the contact zone of the subspecies; away from the contact zone both subspecies had groups of lower sucrose populations. The data show that significant variability exists in this biochemical character, and the pattern of variation is clearly related to the geographic distribution of the populations. The genetic and selective mechanisms involved are unknown.     

Vegetation changes in a shallow African Lake: Response of the vegetation to a recent dry period

This paper shows how the bed of Lake Chilwa in Malawi was colonised by plants during a recent dry period. Aeschynomene pfundii and Diplachne fusca were the principal colonising species on the alkaline mud flats. The subsequent changes in the vegetation after the lake refilled are described. Aeschynomene pfundii survived the flooded conditions for four years and eventually died out as seed germination could not occur underwater. The salinity of the lake during the low water and drying period prevented the lake bed from being extensively colonised by Typha domingensis.     

Isolation of polymorphic microsatellite loci from the red‐billed gull (Larus novaehollandiae scopulinus) and amplification in related species

We describe the isolation and characterization of seven dinucleotide microsatellite loci developed from the red‐billed gull (Larus novaehollandiae scopulinus). Locus‐specific primers were used to genotype individuals from 13 populations of this subspecies as well as individuals from closely related subspecies from Australia and New Caledonia. The primers were tested successfully on other species of gulls and shorebirds. The number of alleles observed within the red‐billed gull ranged from three to 17, and observed heterozygosity varied from 0.359 to 0.787. These microsatellites are likely to be useful for studies of mating systems and population genetics in a wide range of gull species.     

Genetic divergence between Melipona quadrifasciata Lepeletier (Hymenoptera,Apidae) populations

Melipona quadrifasciata is a stingless bee widely found throughout the Brazilian territory, with two recognized subspecies, M. quadrifasciata anthidioides, that exhibits interrupted metasomal stripes, and M. quadrifasciata quadrifasciata, with continuous metasomal stripes. This study aimed to estimate the genetic variability of these subspecies. For this purpose, 127 colonies from 15 Brazilian localities were analyzed, using nine species-specific microsatellite primers. At these loci, the number of alleles ranged from three to 15 (mean: 7.2), and the observed heterozygosity (H_o) ranged from 0.03–0.21, while the expected heterozygosity (H_e) ranged from 0.23–0.47. The genetic distances among populations ranged from 0.03–0.45. The F_ST multilocus value (0.23) indicated that the populations sampled were structured, and the clustering analysis showed the formation of two subgroups and two more distant populations. The first group contained the subspecies M. quadrifasciata quadrifasciata, and the other, the subspecies M. quadrifasciata anthidioides and the two M. quadrifasciata populations with continuous metasomal stripes from northern Minas Gerais. These results confirmed that the yellow metasomal stripes alone are not a good means for correctly identifying the different subspecies of M. quadrifasciata.     

Phylogeography of a salmonid fish,masu salmon Oncorhynchus masou subspecies-complex,with disjunct distributions across the temperate northern Pacific

1. Climate oscillations during the Pleistocene had profound effects on the evolutionary history of freshwater fishes now distributed across northern temperate regions. The extent of continental glaciation on the western side of the North Pacific, including areas of East Asia, was more limited as compared with regions of North America, Europe and high-latitude areas of the North Pacific. Therefore, the effects of climate oscillations might have influenced species in dissimilar ways depending on the species' distribution.
2. We used mitochondrial DNA (mtDNA) and microsatellite DNA (msDNA) markers to clarify the evolutionary history of masu salmon Oncorhynchus masou subspecies-complex (family Salmonidae) distributed in historically non-glaciated regions in the western North Pacific.
3. No marked regional or subspecies-specific mtDNA haplotype associations were recognised, except for O. masou subsp., a lacustrine form endemic to Lake Biwa, an ancient lake in central Honshu. The landlocked subspecies O. masou formosanus, with a disjunct distribution on Taiwan Island, exhibited no diagnostic population features differing from the other subspecies, in either mtDNA or msDNA markers. Mismatch distribution and Bayesian skyline plot analyses indicated relatively recent range expansion and rapid population growth for masu salmon during the last glacial period (c. 0.1–0.15 Ma).
4. Contrary to the mtDNA genealogy, Bayesian clustering using msDNA showed two main genetic clusters, mainly northern populations of the subspecies O. m. masou and populations of the subspecies O. m. ishikawae in southern areas of the Japanese Archipelago. Notably, O. m. formosanus on Taiwan Island was included in the O. m. masou group, and O. masou subsp. was included in the O. m. ishikawae group.
5. Our results suggest that the masu salmon subspecies-complex in historically non-glaciated regions of the Temperate Northern Pacific is characterised by weak population structuring and shallow genetic differentiation among the subspecies, except for O. masou subsp. owing to its long isolation in Lake Biwa. Incomplete lineage sorting and historical inter-subspecies hybridisation, possibly due to secondary contact, seem to be plausible explanations for discrepancies in the mitochondrial DNA genealogy and nuclear DNA genetic structure.

     

Three genetically divergent lineages of the Oryx in eastern Africa: Evidence for an ancient introgressive hybridization

Phylogeographic and population genetic studies using sequence information are frequently used to infer species boundaries and history; and to assess hybridization and population level processes. In this study, partial mitochondrial DNA (mtDNA) control region (423 bp) and cytochrome b sequences (666 bp) of Oryx beisa sampled from five isolated localities in its entire current range in Africa were analyzed to investigate the extent of genetic variation and differentiation between populations. We observed high nucleotide diversity at the control region in the total sample (6.3%) but within populations, it varied considerably ranging from 1.6% to 8.1%. Population pairwise genetic differentiation was generally significantly high (ranging from F _ST = 0.15, P<0.01 to F _ST = 0.54, P<0.001). In the total sample, 29 and 12 haplotypes were observed in the control region and the cytochrome b data sets respectively. For both data sets, the haplotypes cluster into three distinct clades (sequence divergence ranged from 6.0%–12.9% to 0.8%–1.0% for the control region and cytochrome b sequences, respectively) that do not correspond to sampling locations. Two of these clades are found in the same localities (Samburu and Marsabit), which represent the O.beisa beisa subspecies, whereas the last clade represents the fringe-eared oryx (O. beisa callotis). We interpret these findings in terms of an ancient hybridization and introgression between two formerly isolated taxa of Oryx beisa.     

Some biological aspects and life history strategies of Nile tilapia Oreochromis niloticus (L.) in Lake Victoria, Kenya

The life history characteristics of introduced Nile tilapia (Oreochromis niloticus L.) in Lake Victoria, including, sex ratio, fecundity, reproduction, weight‐length relationship and body condition were studied and compared with those of other populations. Samples were collected by trawling and seining in the Kenyan sector of Lake Victoria between June 1998 and December 2000. Males predominated over females (sex ratio 1.42 : 1 : 00). O. niloticus spawned throughout the year but with a peak between December and June. Length at first maturity was (mean ± SD) 30.81 ± 0.09 for females and 34.5 ± 6 0.48 for males. There was little seasonal variation in relative condition, which ranged from 0.92 to 1.05 in males and 0.94 to 1.07 in females. Gonadosomatic index (GSI) was low during the postspawning period (July to October) and high during the protracted breeding period (December and June). Fecundity ranged from 905 to 7619 oocytes for fish of 28 to 51 cm total length (TL) respectively. The relationships between fecundity (F) and total length (L), weight (W) and ovary weight (OW) were: F = 8.159L^1.53, F = 96.269W^0.4504, F = 1806 + 39.4OW. The slope b of the weight‐length relationship was 3.08–3.32 for males and 3.07–3.22 for females. Growth was allometric in both cases and was significantly different from the expected value of 3. The life history strategy of O. niloticus is discussed in context of environmental changes occurring in the lake.     

Bi‐parentally inherited species‐specific markers identify hybridization between rainbow trout and cutthroat trout subspecies

Eight polymerase chain reaction primer sets amplifying bi‐parentally inherited species‐specific markers were developed that differentiate between rainbow trout (Oncorhynchus mykiss) and various cutthroat trout (O. clarki) subspecies. The primers were tested within known F₁ and first generation hybrid backcrosses and were shown to amplify codominantly within hybrids. Heterozygous individuals also amplified a slower migrating band that was a heteroduplex, caused by the annealing of polymerase chain reaction products from both species. These primer sets have numerous advantages for native cutthroat trout conservation including statistical genetic analyses of known crosses and simple hybrid identification.     

Development of microsatellite markers in heterostylous Hedyotis chrysotricha (Rubiaceae)

? Premise of the study: Microsatellite primers were developed for a heterostylous herb, Hedyotis chrysotricha to investigate the effect of habitat fragmentation on its genetic diversity and population structure. ? Methods and Results: Twelve primer sets were developed and their polymorphisms were tested on 47 individuals from two island populations of H. chrysotricha in Thousand Island Lake of China. The number of alleles per locus ranged from five to 10, with an average of seven alleles. Expected heterozygosity per locus ranged from 0.284 to 0.821 and observed heterozygosity ranged from 0.191 to 0.851. ? Conclusions: We showed that all of the 12 microsatellite markers developed for H. chrysotricha are polymorphic within populations, which should provide a powerful tool for assessing population structure and genetic diversity across fragmented and continuous populations, and for studying the genetic effects of habitat fragmentation on this species.