Habitat specialization and sensitivity to change in a threatened crayfish occupying upland streams

1. Crayfish are culturally and ecologically important species in freshwater ecosystems, but many are now threatened with extinction, due to threats such as habitat loss. Depending on their habitat selectivity and sensitivity, freshwater crayfish could provide effective targets for stream monitoring, restoration and conservation. This study used repeated field surveys of the threatened Murray crayfish, Euastacus armatus (family Parastacidae), to establish their habitat preferences and sensitivity to habitat loss across mesohabitat and microhabitat scales within upland streams of south‐east Australia. 2. Selectivity analysis revealed that upland populations of E. armatus displayed a strong preference for areas of intermediate water flow velocity, deeper water, and stream beds dominated by boulders and gravel. Variations in E. armatus density among and within the two upland streams aligned with these microhabitat preferences, with best‐subsets analyses revealing that E. armatus abundance hotspots were best predicted by models combining water depth with percentage cover of boulders, gravel and overhanging riparian vegetation. 3. Major shifts in stream habitat condition from 2009 to 2015 (e.g. 32% and 50% decline in boulder and overhanging vegetation cover, respectively) was associated with a 91% decline in E. armatus mean density within glide‐pool mesohabitats. 4. These findings highlight how habitat specialization in threatened crayfish such as E. armatus may render them sensitive to changes in stream habitat condition, and how targeted monitoring, restoration and conservation of their preferred habitats could yield multiple benefits for montane freshwater ecosystems. Copyright © 2016 John Wiley & Sons, Ltd.     

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.     

Prevalence,geographic distribution and phenotypic differences of Piscirickettsia salmonis EM‐90‐like isolates

Early reports accounted for two main genotypes of Piscirickettsia salmonis, a fish pathogen and causative agent of piscirickettsiosis, placing the single isolate EM‐90 apart from the prototypic LF‐89 and related isolates. In this study, we provide evidence that, contrary to what has been supposed, the EM‐90‐like isolates are highly prevalent and disseminated across Chilean marine farms. Molecular analysis of 507 P. salmonis field isolates derived from main rearing areas, diverse hosts and collected over 6 years, revealed that nearly 50% of the entire collection were indeed typed as EM‐90‐like. Interestingly, these isolates showed a marked host preference, being recovered exclusively from Atlantic salmon (Salmo salar) samples. Although both strains produce undistinguishable pathological outcomes, differences regarding growth kinetics and susceptibility to the antibiotics and bactericidal action of serum could be identified. In sum, our results allow to conclude that the EM‐90‐like isolates represent an epidemiologically relevant group in the current situation of piscirickettsiosis. Based on the consistency between genotype and phenotype exhibited by this strain, we point out the need for genotypic studies that may be as important for the Chilean salmon industry as the continuous surveillance of antimicrobial susceptibility patterns.     

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish <Emphasis Type="Italic">Hoplias malabaricus</Emphasis> hepatocytes

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 μg l^?1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.     

Optimal dietary carbohydrate‐to‐lipid ratios for silvery‐black porgy (Sparidentex hasta) juveniles

Four isonitrogenous diets containing different carbohydrate:lipid (CHO:L) ratios (0.3, 0.6, 1.1 and 1.8) were tested in triplicate groups (16 fish per replicate) of silvery‐black porgy juveniles for 8 weeks. Growth performance was not affected by different dietary CHO:L ratios (P > 0.05); however, the viscerosomatic index, the intraperitoneal fat, whole‐body lipid, energy and n‐3 long chain polyunsaturated fatty acids levels increased with decreasing dietary CHO:L ratios (P < 0.05). Fish fed with 1.8 CHO:L diet had the lowest apparent digestibility coefficients of protein and lipid, as well as the lowest plasma haemolytic and lysozyme activities (P < 0.05). Red blood cell counts and plasma glucose levels were higher in fish fed with 1.1 and 1.8 CHO:L ratio diets than in the other groups (P < 0.05). Plasma alkaline phosphatase and alanine aminotransferase, as well as the level of lipid peroxidation and total antioxidant capacity in the liver and plasma increased as dietary CHO:L ratios decreased (P < 0.05). The results of the current study indicated that the diets with CHO:L ratios between 0.6 and 1.1 are optimal for silvery‐black porgy, whereas higher ratios may result in hyperglycaemia and immune suppression, and lower CHO:L ratios may lead to oxidative stress and liver dysfunction.     

Choline supplementation increased total body lipid gain,while surplus methionine improved growth and amino acid retention in adult Atlantic salmon (Salmo salar)

Methionine–choline‐deficient (MCD) mammals are known to accumulate liver TAG probably due to phosphatidylcholine (PC) deficiency and thus assembly of VLDL and transport of lipids from liver to peripheral organs. To assess whether supplementation of choline could spare methionine and secure a healthy liver metabolism, by reducing the endogenous PC synthesis without interfering with lipid transport and distribution, Atlantic salmon with initial BW of 700 g were fed adequate (1.9 g Met/16 gN) or surplus methionine (2.5 g Met/16 gN) diets of which were supplemented with choline or not for a period of 19 weeks. Fish fed the low‐methionine diets had reduced growth (p = .013) due to reduced protein gain (p = .007), while lipid gain slightly improved in fish fed the choline‐supplemented diets (p = .047). Also, feed conversion improved when fed surplus methionine (p < .001), while choline supplementation had no impact on feed conversion. No interaction between choline and methionine on growth performance or retention existed. Phospholipid status in liver and muscle was not affected by treatments, and no liver TAG accumulation occurred at the methionine levels used. Gene expression of ApoB100 necessary for assembling VLDL or pemt necessary for endogenous PC synthesis was unaffected by treatments. Capacity of methylation (MAT, BHMT) within the liver was not affected by treatment nor was the gene expression of enzymes in liver transsulfuration (CBS or CDO). Methionine status within liver was unaffected by treatments, while free methionine reduced in those fish fed the low‐methionine diets in muscle and plasma. Cystathionine and taurine were elevated when fed surplus methionine. Choline supplementation had no impact on sulphur amino acid metabolites in either tissue. Neither did choline supplementation improve TAG mobilization from liver to muscle. To conclude, choline does not improve endogenous phospholipid synthesis or transport of TAG from liver to muscle depot when added to diets containing 1.9 g Met/16 gN, while surplus methionine improved growth and protein retention, indicating that 1.9 g Met/16 gN is enough to support a healthy liver metabolism, but too low to support muscle protein deposition in adult salmon fed high plant protein diets for longer periods of time.     

Isolation of infectious microalga Prototheca wickerhamii from a carp (Cyprinus carpio) – a first confirmed case report of protothecosis in a fish

Protothecosis is a rare infection caused by environmentally ubiquitous achlorophyllic microalgae of the genus Prototheca. Here, we describe a first case of protothecosis in a carp (Cyprinus carpio), which is at the same time the first case of protothecosis in a fish, confirmed by phenotype‐ and molecular‐based methods, including PCR sequencing of the rDNA cluster and protein profiling using matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry.     

Effect of dietary protein content on growth rate,survival and body composition of juvenile cauque river prawn,Macrobrachium americanum (Bate 1868)

We determined the effect of four concentrations of dietary crude protein, 30.7, 37.2, 41.8 and 46.8% on growth rate, survival and body composition of the juvenile cauque river prawn (Macrobrachium americanum). The prawns were hatched in the laboratory from the spawn of one wild ovigerous female. Prawns consuming 37.2% crude protein reached a final weight of 0.58 g (feed conversion ratio of 2.15), which was significantly better than the other treatments. Survival was 100% in all treatments. Protein content in the diets had no significant effect on whole body proximate composition and amino acid profile. Juveniles consuming the 37.2% crude protein diet grew faster than those fed the other diets. Specific growth rate was adjusted to the two‐slope broken‐line regression analysis model to estimate the optimal protein requirement. In conclusion, these results demonstrated that the 37.2% protein level diet is optimal for juvenile cauque river prawn M. americanum in the experimental conditions of this study.     

Understanding farmers’ potato production practices and use of improved varieties in Chencha,Ethiopia

This study was carried out to better understand non-adoption of improved varieties of potato (Solanum tuberosum) and associated technologies by smallholder farmers in Chencha, Ethiopia. Data were collected through a survey (n = 47) and in-depth interviews (n = 20). It shows how wealth status was a factor of major importance. Most wealthy and some medium-wealthy farmers adopted improved potato varieties and many of the improved production practices; they had access to seed, associated knowledge and support, and sufficient resources that were necessary to apply the improved practices. Non-adoption was common among many medium-wealthy and most poor farmers: they lacked—next to access to the technologies and knowledge—cash, land, and labor. Results indicated the need to rethink research and intervention efforts. Next to paying attention to differences in the access to technology and the related knowledge, there is a need to consider the variation in technology needs, supporting microcredit services, and room to experiment. As a result, different combinations of improved production practices may be adopted.     

Cyprinid herpesvirus 3 as a potential biological control agent for carp (Cyprinus carpio) in Australia: susceptibility of non‐target species

Carp (Cyprinus carpio L.) is a pest species in Australian waterways, and cyprinid herpesvirus 3 (CyHV‐3) is being considered as a potential biological control (biocontrol) agent. An important consideration for any such agent is its target specificity. In this study, the susceptibility to CyHV‐3 of a range of non‐target species (NTS) was tested. The NTS were as follows: 13 native Australian, and one introduced, fish species; a lamprey species; a crustacean; two native amphibian species (tadpole and mature stages); two native reptilian species; chickens; and laboratory mice. Animals were exposed to 100–1000 times the approximate minimum amount of CyHV‐3 required to cause disease in carp by intraperitoneal and/or bath challenge, and then examined clinically each day over the course of 28 days post‐challenge. There were no clinical signs, mortalities or histological evidence consistent with a viral infection in a wide taxonomic range of NTS. Furthermore, there was no molecular evidence of infection with CyHV‐3, and, in particular, all RT‐PCRs for viral mRNA were negative. As a consequence, the results encourage further investigation of CyHV‐3 as a potential biocontrol agent that is specific for carp.