Chris Wood

MOLECULAR GENETIC EVIDENCE FOR PARALLEL LIFE-HISTORY EVOLUTION WITHIN A PACIFIC SALMON (SOCKEYE SALMON AND KOKANEE, ONCORHYNCHUS NERKA)

The Pacific salmon Oncorhynchus nerka typically occurs as a sea‐run form (sockeye salmon) or may reside permanently in lakes (kokanee) thoughout its native North Pacific. We tested whether such geographically extensive ecotypic variation resulted from parallel evolutionary divergence thoughout the North Pacific or whether the two forms are monophyletic groups by examining allelic variation between sockeye salmon and kokanee at two minisatellite DNA repeat loci and in mitochondrial DNA (mtDNA) Bgl II restriction sites. Our examination of over 750 fish from 24 populations, ranging from Kamchatka to the Columbia River, identified two major genetic groups of North Pacific O. nerka: a “northwestern” group consisting of fish from Kamchatka, western Alaska, and northwestern British Columbia, and a “southern” group consisting of sockeye salmon and kokanee populations from the Fraser and Columbia River systems. Maximum‐likelihood analysis accompanied by bootstrapping provided strong support for these two genetic groups of O. nerka; the populations did not cluster by migratory form, but genetic affinities were organized more strongly by geographic proximity. The two major genetic groups resolved in our study probably stem from historical isolation and dispersal of O. nerka from two major Wisconsinan glacial refugia in the North Pacific. There were significant minisatellite DNA allele frequency differences between sockeye salmon and kokanee populations from different parts of the same watershed, between populations spawning in different tributaries of the same lake, and also between sympatric populations spawning in the same stream at the same time. MtDNA Bgl II restriction site variation was significant between sockeye salmon and kokanee spawning in different parts of the same major watershed but not between forms spawning in closer degrees of reproductive sympatry. Patterns of genetic affinity and allele sharing suggested that kokanee have arisen from sea‐run sockeye salmon several times independently in the North Pacific. We conclude that sockeye salmon and kokanee are para‐ and polyphyletic, respectively, and that the present geographic distribution of the ecotypes results from parallel evolutionary origins of kokanee from sockeye (divergences between them) thoughout the North Pacific.

Genetic differentiation in gill raker number and length in sympatric anadromous and nonanadromous morphs of sockeye salmon, Oncorhynchus nerka

The genetic and environmental basis for polymorphism in gill raker number and length in sympatric anadromous and nonanadromous morphs of sockeye salmon, Oncorhynchus nerka, was investigated. Analysis of 30 full sib families involving pure types and reciprocal hybrids revealed that the variation was partitioned significantly among families within cross types and among cross types in both traits. As in the wild, kokanee displayed more gill rakers than sockeye; reciprocal hybrids displayed intermediate counts. Gill raker length also varied markedly among cross types, with pure sockeye displaying 19% longer gill rakers than comparable sized kokanee. This difference was in the opposite direction predicted, given the common positive association between gill raker number and length in sympatric morphs of the same species in fishes. Gill raker number and length were generally not correlated within cross types, suggesting independent divergence of the traits. The results are discussed in relation to genetic and trophic divergence of the morphs and to factors selecting for differentiation in the two gill raker traits.

Implications of Rice Agriculture for Wild Birds in China

Abstract. Data on wild birds in rice fields in China are scarce. The potential significance of Chinese rice fields, which represent about 6% of the worlds wetland area, is considerable but whether this potential is met is largely unknown. In this review, traditional and modern Chinese rice agriculture are compared, including detailing historical changes and their implications for wild birds. Traditional practices, with one crop each year and long periods of fallow flooding, provide greater benefit to biodiversity and species such as the Crested Ibis (Nipponia nippon). The method and alternatives, such as rice-fish, duck-rice and swidden agriculture, are contrasted with modern techniques which, through associated water regimes and chemical use, have been implicated in the decline of biodiversity and of species such as the Black-faced Spoonbill (Platalea minor). Agrochemical use is particularly pertinent because China is likely to have been the worlds largest pesticide consumer since the mid-1990s, with use greatest in rice (Oryza sativa). However, few studies have measured the direct effects of agro-chemicals on wild birds in China. The most detailed information on birds in Chinas rice fields comes from charismatic species such as the Crested Ibis and Red-crowned Crane (Grus japonensis). Preliminary data from possibly the first systematic bird survey of a Chinese rural county are presented. More detailed and widespread studies of the implications of rice agriculture to wild birds in China are required.

Ecological interactions between juveniles of reproductively isolated anadromous and non-anadromous morphs of sockeye salmon, Oncorhynchus nerka, sharing the same nursery lake

Sockeye salmon and kokanee, the anadromous and non-anadromous morphs of Oncorhynchus nerka, spawn in close physical proximity in tributaries to Takla Lake, British Columbia but are reproductively isolated and genetically distinct. Using genetic markers, we were able to investigate, for the first time, ecological interactions between the morphs as juveniles sharing the same nursery lake. Trawl and hydroacoustic surveys conducted in August of 1988 and 1991 revealed that juvenile O. nerka were distributed fairly evenly throughout Takla Lake with average densities ranging from 351–558 fish ha-1 in the north arm to 585–769 fish ha-1 in the west arm. Sockeye salmon were predominant (71–75%) in the west arm whereas kokanee were predominant (82%) in the north arm, a difference attributed to the distribution of spawners in the brood years studied. Within arms, the morphs were intermixed with no detectable difference in relative abundance by depth or among trawl catches. Both morphs were highly selective in their diet, especially in the north arm where fish densities and grazing pressure were lower. As age 0 juveniles, sockeye salmon were significantly larger than kokanee (53 vs. 39 mm on average) but their food habits were virtually identical. Thus we found no evidence of behaviour that would reduce niche overlap between these incipient species.

Prevalence of a Septicemia Disease in the Crested Ibis (Nipponia nippon) in China

Abstract This study investigated six cases of septicemia in young crested ibises (Nipponia nippon). These birds all died with similar clinical signs, including sudden death, anorexia, diarrhea, and lameness. Immediately after death, the birds were necropsied; a blood sample was taken from heart and tissues were sampled from liver, lung, spleen, peritoneal mucus, and feces for bacteriologic examination. Anatomic observation showed that the main findings common to the sick birds were arthrocele, associated with congestion in the femur, tibiotarsus, and ventral side; swelling in the liver; hemorrhagic pericarditis; miliary tubercles in lung; and fibrous tubercles in the synovial capsule of the knee joint with suppurative abscesses. Through bacterial examination, the colonial type of Escherichia coli strain was represented prominently in cultures of the feces, heart blood, liver, lung, spleen, suppurative mucus of the synovial capsule, and peritoneal exudate. These symptoms suggested that the death of a number of endangered crested ibis within a short period was evidence of septicemia. The bacterial inoculation tests were also conducted using domestic pigeon, native chicken, and mice for the presence of and infection with E. coli. The study provided indications of the possible role of E. coli strains as bird pathogens and a potential risk in endangered species. Further work is needed to characterize E. coli strains and the toxin production in this bird. This disease occurrence also adds a note of caution to the continued efforts and interest in the reintroduction of the ibis back into its former wild ranges to ensure that formerly captive individuals do not transmit disease to the wild populations of its own or other sympatric species.

Expression of rat uterine serine proteinases homologous to mouse implantation serine proteinase 2.

Implantation serine protease (ISP) was first identified in the uteri of pregnant mice. It is thought that ISP may have an important role in the initiation of implantation. However, the expression status and detailed functions of ISP remain unclear. In this study, the expression of ISP was investigated in the rat uterus. The analysis of two rat genes registered in GenBank, accession nos. XM_220240 and XM_577076, exhibited high identities to the mouse ISP2 genes, respectively at an mRNA level. We labeled the former as rISP2a and the latter as rISP2b. Using RT-PCR, we found that both genes were expressed in the uterus. Specifically, rISP2a mRNA was detected in the uterus throughout pregnancy, whereas rISP2b mRNA was only expressed in the uterus from day 5 of pregnancy until the end of gestation. Expression of both genes was observed specifically within the endometrial gland epithelium. Furthermore, rISP2a was also observed to be expressed in the fetus and placenta, whereas rISP2b expression was observed in the fetus but not in the placenta. An expressional signal of the rISP2a gene was observed in the spongiotrophoblasts, giant cells and decidual endometrium in the placenta. In the embryo, the ventral specific region was positive in rISP2a and rISP2b gene expression. These findings indicate the possibility that the presently examined genes with high identity to mouse ISP2 may play some role not only during the implantation phase, but also in the development of the placenta and embryo.

New perspectives on habitat selection by the Black-faced Spoonbill Platalea minor based upon satellite telemetry

In 2011 the Black-faced Spoonbill Platalea minor census recorded a 22% drop in numbers from 2010, particularly at the known large coastal wintering sites. During this period, we discovered two new inland wintering sites for the species using satellite telemetry data, one located in China, where the individual followed the Yangtze river as far as Wuhan (500 km inland), and the other across the Vietnam-Cambodia border (70–200 km inland). Long periods of concentrated use of various freshwater habitats were in evidence for these two tagged individuals (China: 79 days, Vietnam/Cambodia: 91 Days) and in the latter case visual confirmation indicated a larger group of at least five individuals. The importance and potential of inland freshwater environments is highlighted towards the further conservation of the recovering population.

Population genetic structure, diversity and stocking effect of the oriental weatherloach (Misgurnus anguillicaudatus) in an isolated island

Genetic endemism of island organisms and the threat to such organisms provided by artificially introduced genes are aspects of major interest in evolutionary and conservation studies of fishes. In this paper the genetic population structure of the oriental weatherloach, Misgurnus anguillicaudatus, in Sado Island of Japan was elucidated by phylogeographic analysis based on partial mitochondrial control region sequences. The specimens were sampled at 62 sites in Sado Island and 14 sites on the mainland close to the island. We found various haplotypes of different origins, most of which had already been reported from the mainland and other places of Japan. This suggests that the loach has been historically introduced to the island from various regions of Japan. Of the 62 sites on the island, cultured/nonnative individuals were confirmed to have been stocked at eight specific sites for feeding of re-introduced Japanese crested ibis (Nipponia nippon). By a Mantel test, geographical and genetic distances were not significantly correlated among the local populations in Sado Island. However a significant correlation was found when the eight stocked local populations were excluded from the analysis. This implied that the genetic distribution pattern of the loach on the island has been disturbed by the stocking. In addition, the nucleotide diversity values of stocked local populations were significantly higher than those of other local populations, also a likely outcome of the stocking. In conclusion, the loach on the island likely had their origins in multiple historical introductions and colonizations, where more recent stocking for the ibis has caused further genetic disturbance to their local populations.

Effects of ectopic expression of human telomerase reverse transcriptase on immortalization of feather keratinocyte stem cells

Normal somatic cells possess a finite life span owing to replicative senescence. Telomerase functions as a potential regulator of senescence in various cells. Expression level of human telomerase reverse transcriptase (hTERT) is correlated with telomerase activity and cellular immortalization. In this study, we investigated the effects of ectopic expression of hTERT on proliferation potential of chicken feather keratinocyte stem cells (FKSCs). We established FKSCs transduced with hTERT catalytic subunit fused with EGFP marker gene (hTERT-EGFP-FKSCs). hTERT-EGFP-FKSCs had the great potential of proliferation in vitro and expressed kerainocyte stem cell markers integrin beta1 and CD49c. Keratin 15 and keratin 19, as native FKSCs, were also detected in hTERT-EGFP-FKSCs. By the analysis of fluorescent RT-PCR, western blotting and TRAP assay, hTERT-EGFP-FKSCs were positive for telomerase activity, in comparison with native FKSCs showing no telomerase activity. We demonstrated that ectopic expression of hTERT could result in immortalization of FKSCs. Tumorigenecity of hTERT-EGFP-FKSCs were examined by soft agar assay and transplantation into NOD-SCID mice. Results showed that hTERT-EGFP-FKSCs sustained the cellular characteristics of native FKSCs and had no transforming activity. In vivo differentiation multipotentials of hTERT-EGFP-FKSCs were confirmed by transplantation into developing chicken embryos and in situ hybridization analysis. These data provide a novel framework for understanding human telomerase activity in different species and suggest a new insight for manipulating hTERT for therapeutic purposes in treating tissue injury and aging.

Distribution pattern of loaches (teleostei: Cobitoidea) in the river east Tiaoxi, China

Abstract. The distribution patterns of loaches (Teleostei: Cobitoidea) were revealed in the River East Tiaoxi in China. Eight loach species (Cobitis sinensis, C. dolichorhynchus, C. laterimaculata, Misgurnus anguilicaudatus, Paramisgurnus dabryanus, Leptobotia tchangi, Vanmanenia pingchowensis and V. stenosoma) were found in field surveys at 90 sites. A macroscale analysis of the geographical distribution showed that related species were segregated in accordance with longitudinal alterations along the river course. M. anguillicaudatus was widely distributed from the middle reaches to upstream of the river whereas the related species, P. dabryanus, was distributed only downstream. Three spined loaches, namely C. dolichorhynchus, C. sinensis, and C. laterimaculata, were distributed in the middle reaches, the middle to upstream reaches, and the upstream section of the river, respectively. The distribution of V. pingchowensis was further upper stream than that of V. stenosoma in the upstream section. More detailed microscale analysis revealed that L. tchangi and the two Vanmanenia species, V. stenosoma and V. pingchowensis were mostly found in the ‘Riffle’ (lotic) habitats, whereas the two spined loaches, C. laterimaculata and C. sinensis preferred the ‘Pool’ (lentic) habitats. These results demonstrated that related loach species preferred similar habitat units in the River East Tiaoxi system.