Bioindicators of homeostasis’ constants of growing conditions of warm-water aquaculture objects in the context of obtaining marketable products

To create recommendations regarding the technological process of growing objects of warm-water aquaculture, the health of individuals in the created conditions of maintain is a necessary study. Taking into account the similarity of the necessary conditions for the organization of growing warm-water aquaculture objects, it is important to monitor the indicators that are bioindicators of homeostasis constants with a projection on the conditions of maintain, for subsequent adjustment of satisfactory conditions of maintain. First of all, these are hematological and biochemical bioindicators that determine the general characteristics of blood in the case of fish objects of sterlet and hemolymph in the case of crustacean objects of Australian freshwater crayfish and the concentration index of total serum protein that determines the “physiological norm”. When growing sterlet, a high level of total blood protein was established at 40.14±1.73 g/l and the hemolymph of the Australian freshwater crayfish was determined to have a protein concentration of 40.8±4.5 g/l. The presented analysis of bioindicators complements the missing information for monitoring the data of representatives of warm-water aquaculture, which will be valuable for specialists engaged in breeding these objects.

Labile sex chromosomes and a novel candidate sex-determination gene in the Australian freshwater fish family Percichthyidae

Sex-specific ecology has management implications, but rapid sex-chromosome turnover in fishes hinders development of markers to sex monomorphic species. Here, we use annotated genomes and reduced-representation sequencing data for two Australian percichthyids, the Macquarie perch Macquaria australasica and the golden perch M. ambigua, and whole genome resequencing data for 50 Macquarie perch of each sex, to detect sex-linked loci, identify a candidate sex-determining gene and develop an affordable sexing assay. In-silico pool-seq tests of 1,492,004 Macquarie perch SNP loci revealed that a 275-Kb scaffold, containing the transcription factor SOX1b gene, was enriched for gametologous loci. Within this scaffold, 22 loci were sex-linked in a predominantly XY system, with females being homozygous at all 22, and males being heterozygous at two or more. Seven XY-gametologous loci were within a 146-bp region. Being ~38 Kb upstream of SOX1b, it might act as an enhancer controlling SOX1b transcription in the bipotential gonad that drives gonad differentiation. A PCR-RFLP sexing assay, targeting one of the Y-linked SNPs, tested in 66 known-sex Macquarie perch and two individuals of each sex of three confamilial species, and amplicon sequencing of 400 bp encompassing the 146-bp region, revealed that the few sex-linked positions differ between species and between Macquarie perch populations. This indicates sex-chromosome lability in Percichthyidae, also supported by non-homologous scaffolds containing sex-linked loci for Macquarie- and golden perches. The resources developed here will facilitate genomic research in Percichthyidae. Sex-linked markers will be useful for determining genetic sex in some populations and studying sex chromosome turnover.

HEMATOLOGICAL AND BIOCHEMICAL INDICATORS OF AUSTRALIAN RED-CLAW CRAYFISH HEMOLYMPH

The article describes Australian red claw crayfish, Cherax quadricarinatus, which is currently one of the most promising cultivation targets. However, to increase production volumes, it is necessary to improve cultivation technologies, which should be based on the data of physiological and biochemical parameters. An objective method for monitoring the physiological state of Australian red claw crayfish in the artificial conditions is determining its hematological and biochemical indicators that complement the general characteristics of hemolymph and later define its physiological norm. When giving the objects the compound feed, which the authors produced by their own recipe, there was recorded a high level of total hemolymph protein of 40.8 ± 4.5 g/l. According to the results of biochemical research, the cholesterol content in the hemolymph of Australian crayfish was 3.2 ± 0.6 mmol/l, and the concentration of lipoproteins was 0.8 ± 0.2 g/l (levels within the reference values). When considering the hemogram of hemolymph there were found the dominant types and linear dimensions of native formed elements. The diameter of agranulocytes in Australian freshwater crayfish varied from 85 to 90 μm, semi-granulocytes - from 90 to 95 μm, granulocytes - from 65 to 78 μm, transparent cells differed by greater variability in size, which ranged from 85 to 120 μm. The part of granulocytes in relation to other shaped elements exceeded 50%, for transparent cells it turned out to be above 20%, for semi-granulocytes - about 15% and for agranulocytes - about 5%, which means that the latter form the rarest group. The established ratios are specific for Cherax quadricarinatus, its hemocytic characteristics greatly differs from Astacus leptodactylus, which is the main object of comparative research. The results of comparative analysis of the cellular composition of the hemolymph of the Australian freshwater crayfish Cherax quadricarinatus (Von Martens, 1868) demonstrate the taxonomic specificity of this representative of freshwater aquaculture

Hydrological, geochemical and land use drivers of greenhouse gas dynamics in eleven sub-tropical streams

Greenhouse gas (GHG) emissions from freshwater streams are poorly quantified in sub-tropical climates, especially in the southern hemisphere where land use is rapidly changing. Here, we examined the distribution, potential drivers, and emissions of carbon dioxide (CO2), nitrous oxide (N2O) and methane (CH4) from eleven Australian freshwater streams with varying catchment land uses yet similar hydrology, geomorphology, and climate. These sub-tropical streams were a source of CO2 (74 ± 39 mmol m−2 day−1), CH4 (0.04 ± 0.06 mmol m−2 day−1), and N2O (4.01 ± 5.98 µmol m−2 day−1) to the atmosphere. CO2 accounted for ~ 97% of all CO2-equivalent emissions with CH4 (~ 1.5%) and N2O (~ 1.5%) playing a minor role. Episodic rainfall events drove changes in stream GHG due to the release of soil NOx (nitrate + nitrite) and dissolved organic carbon (DOC). Groundwater discharge as traced by radon (222Rn, a natural groundwater tracer) was not an apparent source of CO2 and CH4, but was a source of N2O in both agricultural and forest catchments. Land use played a subtle role on greenhouse gas dynamics. CO2 and CH4 increased with catchment forest cover during the wet period, while N2O and CH4 increased with agricultural catchment area during the dry period. Overall, this study showed how DOC and NOx, land use, and rainfall events interact to drive spatial and temporal dynamics of GHG emissions in sub-tropical streams using multiple linear regression modelling. Increasing intensive agricultural land use will likely decrease regional CO2 and CH4 emissions, but increase N2O.

Genomic data shed light on sex-determination in Australian freshwater Percichthyid fish species: Many ways to be a male

Understanding sex-specific biology can aid conservation management. But understanding genomic sex differences of monomorphic fish species and developing molecular sexing assays is challenged by their diverse sex-determination systems. To facilitate research on Percichthyid fish, predominant in the Australian freshwater biota, we report whole genome sequences and annotations of the endangered Macquarie perch Macquaria australasica and its sister species, the golden perch M. ambigua. To identify sex-linked loci, we conducted whole genome resequencing on 100 known-sex Macquarie perch. In-silico pool-seq comparisons revealed few sex differences, but a 275-Kb SOX-containing scaffold was enriched for gametologous loci- homozygous in females, heterozygous in males. Within this scaffold we reconstructed X- and Y-linked 146-bp haplotypes containing 5 sex-linked SNPs, ~38 Kb upstream of SOX, and developed a PCR-RFLP sexing assay targeting the Y-linked allele of one SNP. We tested this assay in a panel of known-sex Macquarie perch, and smaller panels of three other confamilial species. Amplicon sequencing of 400 bp encompassing the 146-bp region revealed that the few sex-linked positions differ interspecifically, and within Macquarie perch such that its sexing test approached 100% reliability only for the populations used in assay development. Similarly, Macquarie- and golden perch genome-wide DArTseq SNPs revealed different sex-linked loci across non-homologous scaffolds. Overall, we identified 22 sex-linked SNPs in Macquarie perch in a predominantly XX/XY system in which females are homozygous at all 22, and males are heterozygous at 2 or more. The resources here will facilitate multi-locus sexing assays for both species and research on Percichthyid biology.