ECOLOGY AND BEHAVIOR OF WILD BLACK-CRESTED GIBBONS (HYLOBATES-CONCOLOR) IN CHINA WITH A RECONSIDERATION OF EVIDENCE FOR POLYGYNY

The black-crested gibbon, Hylobates concolor, is one of the few species of gibbons that has not yet been the subject of a long term field study. Field observations in the Ai Lao and Wu Liang Mountains of Yunnan Province, China indicate that in this area the habitat and ecology of this species differ markedly from those of other gibbons that have been studied to date. These differences are correlated with some behavioral differences. In particular, these gibbons apparently have greater day ranges than other gibbons. It has also been suggested that this species lives in polygynous groups. To demonstrate this requires observation of groups with two or more females with young. Our own observations and those from other recent studies suggest that there are alternative explanations consistent with available data.

REPORT ON THE DISTRIBUTION, POPULATION, AND ECOLOGY OF THE YUNNAN SNUB-NOSED MONKEY (RHINOPITHECUS-BIETI)

The Yunnan snub-nosed monkey (Rhinopithecus bieti), an endangered species in China, has received more protection in theory than in practice. Therefore it is on the very verge of extinction. The population of the species was estimated less than 2,000 individuals spread in 19 distinct groups. It was confirmed that the monkey was confined to the Yunling Mountain System, the area between the Yangtze River (Changjiang, aka Jinshajiang) to the east and the Mekong River (Lancangjiang) to the west. We further concluded that a lowland belt to the east, about 100 km long and 20 - 30 km wide was not suitable habitat for the monkeys, and appeared to serve as the natural ecogeologic barrier for the species. Our results indicated that the southern limit of the distribution was at Longma (26-degrees 14'N), and that the northern limit of the distribution was at Xiaochangdu (29-degrees 20'N). The distribution area of the species was substantially smaller than previously estimated. There were substantial ecological differences between the southern and northern parts of the species range. The monkey was found only in fir-larch forest.

Diet and feeding behavior of Rhinopithecus bieti at Xiaochangdu, Tibet: Adaptations to a marginal environment

The diet and feeding ecology of a wild subpopulation of black-and-white snub-nosed monkeys (Rhinopithecus bieti) were studied at Xiaochangdu in Honglaxueshan Nature Reserve, Tibet. This region is climatologically harsher than any other inhabited by non-human primates. Black-and-white snub-nosed monkeys fed on 48 parts of 25 plant species, at least three species of lichens and seven species of invertebrates. The number of food items exploited varied markedly among seasons, with dietary diversity being greatest in spring and summer. In winter, black-and-white snub-nosed monkeys had to subsist on fallback foods such as dried grass and bark. Ubiquitous lichens formed a major dietary constituent throughout the year, contributing about 75% of feeding records. Even though lichens act as a staple, our findings signify that the monkeys at Xiaochangdu prefer feeding on foliage, which is higher in protein content than the former. We provide evidence that black-and-white snub-nosed monkeys are able to cope with an array of food items other than lichens and hence can be regarded as feeding generalists. We discuss the results with reference to previous studies on other subpopulations living in habitats that are floristically more diverse and offer more plant food items than the marginal habitat at Xiaochangdu.

Industrial ecology at factory level - A conceptual model

Growing environmental concerns caused by natural resource depletion and pollution need to be addressed. One approach to these problems is Sustainable Development, a key concept for our society to meet present as well as future needs worldwide. Manufacturing clearly has a major role to play in the move towards a more sustainable society. However it appears that basic principles of environmental sustainability are not systematically applied, with practice tending to focus on local improvements. The aim of the work presented in this paper is to adopt a more holistic view of the factory unit to enable opportunities for wider improvement. This research analyses environmental principles and industrial practice to develop a conceptual manufacturing ecosystem model as a foundation to improve environmental performance. The model developed focuses on material, energy and waste flows to better understand the interactions between manufacturing operations, supporting facilities and surrounding buildings. The research was conducted in three steps: (1) existing concepts and models for industrial sustainability were reviewed and environmental practices in manufacturing were collected and analysed; (2) gaps in knowledge and practice were identified; (3) the outcome is a manufacturing ecosystem model based on industrial ecology (IE). This conceptual model has novelty in detailing IE application at factory level and integrating all resource flows. The work is a base on which to build quantitative modelling tools to seek integrated solutions for lower resource input, higher resource productivity, fewer wastes and emissions, and lower operating cost within the boundary of a factory unit. © 2012 Elsevier Ltd. All rights reserved.

Industrial ecology at factory level: A prototype methodology

The concept of sustainable manufacturing is a form of pollution prevention that integrates environmental considerations in the production of goods while focusing on efficient resource use. Taking the industrial ecology perspective, this efficiency comes from improved resource flow management. The assessment of material, energy and waste resource flows, therefore, offers a route to viewing and analysing a manufacturing system as an ecosystem using industrial ecology biological analogy and can, in turn, support the identification of improvement opportunities in the material, energy and waste flows. This application of industrial ecology at factory level is absent from the literature. This article provides a prototype methodology to apply the concepts of industrial ecology using material, energy and waste process flows to address this gap in the literature. Various modelling techniques were reviewed and candidates selected to test the prototype methodology in an industrial case. The application of the prototype methodology showed the possibility of using the material, energy and waste resource flows through the factory to link manufacturing operations and supporting facilities, and to identify potential improvements in resource use. The outcomes of the work provide a basis to build the specifications for a modelling tool that can support those analysing their manufacturing system to improve their environmental performance and move towards sustainable manufacturing. © IMechE 2012.

Genomic organization and expression analysis of Toll-like receptor 3 in grass carp (Ctenopharyngodon idella)

Toll-like receptor 3 (TLR3) participates in the innate immune response by recognizing viral pathogens. To investigate grass carp immune system responding to GCRV (grass carp reovirus) infection, the full-length cDNA sequence and genomic organization of grass carp TLR3 (CiTLR3) was identified and characterized. The full-length genome sequence of CiTLR3 is composed of 5668 nucleotides, including five exons and four introns. The full-length of CiTLR3 cDNA is 3681 bp in length and encodes a polypeptide of 904 amino acids with an estimated molecular mass of 102,765 Da and a predicted isoelectric point of 8.35. Analysis of the deduced amino acid sequence indicated that CiTLR3 has four main structural domains, including a signal peptide sequence, 14 LRR (leucine-rich repeat) motifs, a transmembrane region and a TIR (Toll/interleukin-1 receptor) domain. It is most similar to the crucian carp (Carassius auratus) TLR3 amino acid sequence with an identity of 99%. Quantitative RT-PCR analysis showed that CiTLR3 transcripts were significantly up-regulated starting at day 1 and continued through day 7 following GCRV infection (P < 0.05). These data implied that CiTLR3 is involved in antiviral defense, provide molecular and functional information for grass carp TLR3, and implicate their role in mediating immune protection against grass carp viral diseases. (C) 2009 Elsevier Ltd. All rights reserved.

Effect of beta-glucan on activity of antioxidant enzymes and Mx gene expression in virus infected grass carp

The effects of beta-glucan, an immunostimulatory agent, on the superoxide dismutase (SOD) and catalase (CAT) activities of erythrocytes and Mx gene expression were studied from grass carp that were challenged with grass carp hemorrhage virus (GCHV). The SOD and CAT activities in erythrocytes and Mx gene expression in spleen from the fish were detected by spectrophotometry and RT-PCR, respectively. Negative control fish were injected with PBS; positive control groups were injected with either P-glucan or GCHV only; and the experimental groups were pre-injected with beta-glucan 15 days prior to injection with GCHV. The results show that the SOD and CAT activities were higher in fish injected with beta-glucan for 15 days than the negative control group injected with PBS. The SOD and CATactivities significantly decreased when the fish were challenged with GCHV, but it was higher in the group pre-treated with beta-glucan than in infected fish not pre-treated, 15 days after GCHV infection. Mx gene expression levels increased during the early stages (at 12 h and 36 h) of GCHV infection, and it remained at higher levels from the 6th till the 10th day in the beta-glucan pre-treated group, but it was failing from the 6th day in the beta-glucan untreated group. The GCHV-infected group pre-treated with P-glucan had a higher survival rate (60%) than the group not pre-treated with P-glucan (20%), suggesting that beta-glucan possesses or enhances anti-viral responses. (C) 2009 Elsevier Ltd. All rights reserved.

Toll-like receptor 4 signaling pathway can be triggered by grass carp reovirus and Aeromonas hydrophila infection in rare minnow Gobiocypris rarus

Toll-like receptor 4 (TLR4) is critical for LPS recognition and cellular responses. It also recognizes some viral envelope proteins. Detection mostly results in the inflammation rather than specific antiviral responses. However, it's unclear in fish. In this report, a TLR4 gene (named as GrTLR4b) was cloned and characterized from rare minnow Gobiocypris rarus. The full length of GrTLR4b cDNA consists of 2766 nucleotides and encodes a polypeptide of 818 amino acids with an estimated molecular mass of 94,518 Da and a predicted isoelectric point of 8.41. The predicted amino acid sequence comprises a signal peptide, six leucine-rich repeat (LRR) motifs, one leucine-rich repeat C-terminal (LRRCT) motif, followed by a transmembrane segment of 23 amino acids, and a cytoplasmic region of 167 amino acids containing one Toll - interleukin 1 - receptor (TIR) motif. It's closely similar to the zebrafish (Danio rerio) TLR4b amino acid sequence with an identity of 77%. Quantitative RT-PCR analysis showed GrTLR4b mRNA was constitutive expression in gill, heart, intestine, kidney, liver, muscle and spleen tissues in healthy animals and up-regulated by viruses and bacteria. After being infected by grass carp reovirus or Aeromonas hydrophila, GrTLR4b expressions were up-regulated from 24 h post-injection and lasted until the fish became moribund (P < 0.05). These data implied that TLR4 signaling pathway could be activated by both viral and bacterial infection in rare minnow. (C) 2009 Elsevier Ltd. All rights reserved.

Enhanced grass carp reovirus resistance of Mx-transgenic rare minnow (Gobiocypris rarus)

In the interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful. Mx proteins have direct antiviral activity and inhibit a wide range of viruses by blocking an early stage of the viral genome replication cycle. However, antiviral activity of piscine Mx remains unclear in vivo. In the present study, an Mx-like gene was cloned, characterized and gene-transferred in rare minnow Gobiocypris rarus, and its antiviral activity was confirmed in vivo. The full length of the rare minnow Mx-like cDNA is 2241 bp in length and encodes a polypeptide of 625 amino acids with an estimated molecular mass of 70.928 kDa and a predicted isoelectric point of 7.33. Analysis of the deduced amino acid sequence indicated that the mature peptide contains an amino-terminal tripartite GTP-binding motif, a dynamin family signature sequence, a GTPase effector domain and two carboxy-terminal leucine zipper motifs, and is the most similar to the crucian carp (Carassius auratus) Mx3 sequence with an identity of 89%. Both P0 and F1 generations of Mx-transgenic rare minnow demonstrated very significantly high survival rate to GCRV infection (P < 0.01). The mRNA expression of Mx gene was consistent with survival rate in F1 generation. The virus yield was also concurrent with survival time using electron microscope technology. Rare minnow has Mx gene(s) of its own but introducing more Mx gene improves their resistance to GCRV. Mx-transgenic rare minnow might contribute to control the GCRV diseases. (C) 2008 Published by Elsevier Ltd.

Grass carp reovirus activates RNAi pathway in rare minnow, Gobiocypris rarus

Dicer catalyzes the initiation step of RNA interference (RNAi) which is known to play a significant role in innate immune response to viral infection in many organisms. To study the RNAi-related pathway after virus infection in fish, we identified a partial cDNA sequence of dicer from rare minnow, Gobiocypris rants. Real-time quantitative RT-PCR (qRT-PCR) demonstrated the Dicer transcript level was the highest at zygote stage, decreased at prim-5 stage, and was stable from the protruding mouth to adult stage. Regular RT-PCR analysis showed that the Dicer gene expressed widely in the tested tissues, including brain, gill, heart, intestine, kidney, liver, muscle, ovary, spleen and testis. The expression of Dicer mRNA was significantly increased in the early period of Grass carp reovirus (GCRV) infection, and declined from 24 It post-injection (h p.i.) (P<0.05). The mRNA expression returned to control levels at 48 h p.i. (P>0.05). Under transmission electron microscope, virions were difficulty to find out in 12 h p.i., and virus inclusion bodies and few scattered viral particles were easily visualized from 24 h p.i. to moribund. These results implied GCRV triggered the RNAi pathway in the early stages of infection and perhaps virus inclusion bodies suppressed the antiviral functions of RNAi mechanism. (C) 2009 Published by Elsevier B.V.

Ecology of macrozoobenthic communities in two plateau lakes of Southwest China

Ecological studies on macrozoobenthos were conducted in two small plateau lakes in the Yunnan-Guizhou Plateau, Southwest China: Xingyun Lake (XL), a eutrophic lake whose main source of primary production was phytoplankton (Chl a=99.76 +/- 24.01 mu g/L), and Yangzong Lake (YL), a mesotrophic lake. Sampling was carried out from October 2002 to May 2004. Altogether 23 benthic taxa were identified in XL and 21 taxa in YL. The density of benthos in XL was much lower than that in YL, but the biomass was about equal in the two lakes, being 1 423 ind/m(2) and 8.71 g/m(2) in XL and 4 249 ind/m(2) and 8.60 g/m(2) in YL. The dominant species were Limnodrilus hoffmeisteri, Branchiura sowerbyi, Aulodrilus pluriseta and Chironomus sp. in XL and Limnodrilus hoffmeisteri, Aulodrilus pluriseta and Bellamya sp. in YL. Seasonal fluctuation occurred, showing richer species in summer and winter, but the density and biomass varied in different ways in the two lakes. Analyses on functional feeding groups indicate that collector-gatherers were predominant, but the relative abundances of other groups were different. Stepwise multiple regression analysis demonstrated that the water depth, conductivity and chlorophyll a were the key factors affecting macrozoobenthic abundance in the lakes.

Ultrastructural alteration of lymphocytes in spleen and pronephros of grass carp (Ctenopharyngodon idella) experimentally exposed to microcystin-LR

Relatively little is known in relation to pathological changes of immune organs in fish when exposed to MC-LR. The ultrastructural alteration of lymphocytes was examined in the spleen and pronephros of grass carp Ctenopharyngodon idella injected experimentally with microcystin-LR. The fish were intraperitoneally injected with MC-LR at a dose of 50 mu g/kg body weight, and the spleen and pronephros were dissected out at 1, 2, 7, 14 and 21 days post intraperitoneal injection (dpi). Pathological changes were then examined by transmission electron microscopy. Apoptosis was detected only in lymphocytes in the spleen, with obvious apoptotic features observed at 2 dpi; pathological changes of lymphocytes in the pronephros were also serious with mitochondria being highly edematous. However, damaged lymphocytes were almost un-observed in the spleen and pronephros at 21 dpi. These findings suggest that MC-LR can induce toxic effect on immune organs in grass carp, and the spleen may be much more sensitive to MC-LR stimulation. (C) 2008 Elsevier B.V. All rights reserved.

Detection of cutaneous antibodies in excised skin explants from grass carp, Ctenopharyngodon idella (Valenciennes), immune to Scophthalmus maximus rhabdovirus

This study determined whether cutaneous antibodies were present in excised skin explants of grass carp, Ctenopharyngodon idella, immune to Scophthalmus maximus rhabdovirus (SMRV). Culture fluid from immune skin explants were assayed by indirect enzyme-linked immunosorbent assay (iELISA), Western blot, indirect immunofluorescent assay (IFA) and flow cytometry (FCM). iELISA showed that cutaneous antibody titres were much lower (1:12) than antiserum titres (1:1458) from intraperitoneally immunized grass carp. The phosphoprotein and matrix protein antigens of purified SMRV proteins were recognized by cutaneous antibodies from skin culture fluid using Western blot. The skin culture fluid produced staining signals in viral assembly sites and cytoplasm of SMRV-infected epithelioma papulosum cyprini (EPC) cells by IFA. FCM showed that 4.39% SMRV-infected EPC cells were detected, while non-specific reaction was seen in 2% of control cells. This is the first description of cutaneous antibodies against SMRV in grass carp.