Cryopreservation effects on the sperm quality of cachama blanca Piaractus brachypomus (Cuvier 1818)

The effects of straws volume, cryoprotectants and thawing temperatures were evaluated on the sperm quality of cachama blanca Piaractus brachypomus (Cuvier), an important Colombian fish species. Sexually mature fish were induced to ovulation or spermiation with a carp pituitary extract. A pool of suitable sperm samples was diluted in glucose, egg yolk, dimethyl sulphoxide (DMSO‐10%), methanol (MET‐10%) or ethylene glycol (ETG‐5%) and packed in 0.5, 2.5 or 5.0 mL straws and frozen in nitrogen vapour. The thawing process was performed in a 35 or an 80 °C water bath. The fertility was evaluated after 6 h post fertilization. The highest motility percentage (33 ± 3%) was observed with sperm cryopreserved with DMSO, packed in 5 mL straws and thawed at 35 °C. The treatments with DMSO and MET packed in 0.5 and 5.0 mL straws and thawed at 35 °C showed the highest fertility (higher than 71%) and the lowest fertility was obtained with MET‐2.5 mL (9 ± 5%). In all the treatments, a significant decrease in the sperm quality was observed at 80 °C. Sperm cryopreserved with DMSO‐10% or MET‐10%, packed in 2.5 or 5.0 mL straws are suitable to achieve acceptable fertilization and to fertilize high amounts of eggs.     

Multilocus sequence typing detects new Piscirickettsia salmonis hybrid genogroup in Chilean fish farms: Evidence for genetic diversity and population structure

Piscirickettsia salmonisis the causative bacterial pathogen of piscirickettsiosis, a salmonid disease that causes notable mortalities in the worldwide aquaculture industry. Published research describes the phenotypic traits, virulence factors, pathogenicity and antibiotic‐resistance potential for various P. salmonisstrains. However, evolutionary and genetic information is scarce for P. salmonis. The present study used multilocus sequence typing (MLST) to gain insight into the population structure and evolution of P. salmonis. Forty‐two Chilean P. salmonisisolates, as well as the type strain LF‐89^T, were recovered from diseased Salmo salar, Oncorhynchus kisutchand Oncorhynchus mykissfrom two Chilean Regions. MLST assessed the loci sequences of dnaK, efp, fumC, glyA, murG, rpoD and trpB. Bioinformatics analyses established the genetic diversity among P. salmonis isolates (H = 0.5810). A total of 23 sequence types (ST) were identified, 53.48% of which were represented by ST1, ST5 and ST2. Population structure analysis through polymorphism patterns showed few polymorphic sites (218 nucleotides from 4,010 bp), while dN/dS ratio analysis indicated purifying selection for dnaK, epf, fumC, murG, and rpoD but neutral selection for the trpB loci. The standardized index of association indicated strong linkage disequilibrium, suggesting clonal population structure. However, recombination events were detected in a group of seven isolates. Findings included genogroups homologous to the LF‐89^T and EM‐90 strains, as well as a seven‐isolate hybrid genogroup recovered from both assessed regions (three O. mykiss and four S. salar isolates). The presented MLST scheme has comparative potential, with promising applications in studying distinct P. salmonis isolates (e.g., from different hosts, farms, geographical areas) and in understanding the epidemiology of this pathogen.     

Using genetic methods for analysis of fish meals and feeds employed in Greek mariculture

Large quantities of high protein fish meals are needed to sustain cultured species and thus the impact to marine ecosystem has been highly discussed. The aim of this study was to apply a PCR‐cloning methodology for a robust insight into the composition of commercial fish meals and feeds for farmed species of the Greek mariculture, assessing the risk posed by aquaculture to marine ecosystems but also the risk posed by commercial fish feeds to the increase in trophic level of species farmed in Greece. 89% of the sequences were identified to species level and only 11% to genus/family level. Overall, a total of 49 taxa were identified (44 fish species/taxon, five non‐fish species/taxon). Even though small pelagic fish like Engraulis sp. were the main portion, a wide range of species constituted the fish meals and feeds. Plant and animal species were also detected as an alternative protein source. Feed products employed in Greek mariculture still contain large portions of fish meals which increase the mean trophic level of farmed species causing a farming up trend. The results emphasize that such molecular methodologies are needed to certify aquafeeds allowing fish feed producers to demonstrate their commitment to sustainable aquaculture.     

Soy protein concentrate as a substitute for fish meal in diets for juvenile Acanthopagrus schlegelii: effects on growth,phosphorus discharge and digestive enzyme activity

The current high demand and cost of fish meal (FM) necessities the evaluation of alternative plant protein ingredients in diets of farmed marine fish. A 56‐day feeding trial was performed to study the effects of replacement of FM with soy protein concentrate (SPC) in diets of Acanthopagrus schlegelii. Diets were prepared at levels of 0%, 30%, 47.5%, 65%, 82.5% and 100% SPC, respectively, replacing FM. The results indicated no significant differences (p > .05) in % weight gain (WG) and specific growth rate of fish fed S30% to S82.5% diets compared with the control diet while, further inclusion at 100% significantly depressed growth performance. SPC inclusion and phytase supplementation significantly affected the phosphorous discharge (P‐load) showing lowest value (3.83 ± 0.53 g/kg WG) in S100% compared to control (14.79 ± 0.37 g/kg WG) and in fish fed S30% diet (13.24 ± 0.89 g/kg WG) (p < .05). The results of this study showed that FM could be substituted up to 82.5% by SPC in the diet of Acanthopagrus schlegelii fingerlings (5.53 ± 0.12 g) without any adverse effects. Phytase supplementation SPC based diets could be effective in reducing the phosphorus load in the aquatic culture environment.     

Effects of Salinity on Biogenic Amines,Hemolymph Osmotic Pressure,and Activity of Gill’s Na+/K+‐ATPase in Charybdis japonica (Crustacea,Decapoda)

The effects of salinity on hemolymph osmotic pressure, gill Na⁺/K⁺‐ATPase activity and dopamine (DA), norepinephrine (NE), serotonin (5‐HT) in the gills, and hemolymph of the adult Charybdis japonica were studied. DA levels increased significantly (P < 0.05), while the NE and 5‐HT revealed contrary change in hemolymph and gills. The iso‐osmotic point of C. japonica (911.4 mOsm/kg) was at salinity of 27.87 ppt. The Na⁺/K⁺‐ATPase activity of gill showed negative correlation with salinity in the hypotonic environment (<27.87 ppt). The results of this experiment indicated that C. japonica had great capability to acclimate to low salinity.     

Effect of dietary protein reduction with lysine and methionnine supplementation on growth performance,body composition and total ammonia nitrogen excretion of juvenile grass carp,Ctenopharyngodon idella

A 63‐day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant‐based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg^?1, 300 g kg^?1 and 280 g kg^?1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day^?1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP‐ and 28CP‐diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P < 0.05), and the interaction between dietary protein level and amino acid supplementation was noted between WG and SGR (P < 0.05). Feed intake (FI) was significantly increased with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05), but feed conversion ratio (FCR) showed a significant decreasing trend (P < 0.05). Two days after total ammonia nitrogen (TAN) concentration test, the values of TAN discharged by the fish 8 h after feeding were 207.1, 187.5, 170.6, 157.3, 141.3 and 128.9 mg kg^?1 body weight for fish fed 32CP, 32AA, 30CP, 30AA, 28CP and 28AA diets, respectively. TAN excretion by grass carp was reduced in plant‐based practical diets with the increase in dietary protein level and the supplementation of lysine and methionine (P < 0.05). The results indicated that lysine and methionine supplementation to the plant protein sources‐based practical diets can improve growth performance and feed utilization of grass carp, and the dietary crude protein can be reduced from 320 g kg^?1 to 300 g kg^?1 through balancing amino acids profile. The positive effect was not observed at 280 g kg^?1 crude protein level.     

CD117‐ and vimentin‐positive telocytes in the bovine teat sphincter

Mastitis is a common economically relevant problem in dairy farming. As the major entry for pathogens is the papillary duct, one of the first defence mechanisms is the teat sphincter. This sphincter shows a rhythmic contractility of yet unknown origin. Searching for possible modulatory pacemaker cells, teat sphincters of eight cows were stained immunohistochemically with antibodies against CD117 and vimentin and evaluated microscopically for the presence of telocytes. CD117‐ and vimentin‐positive telocytes with telopodes were found in close contact with smooth muscle cells. Our findings present a first evidence of telocytes in the teat of bovines.     

Effects of illite supplementation on in vitro and in vivo rumen fermentation,microbial population and methane emission of Hanwoo steers fed high concentrate diets

This study was conducted to evaluate the effects of feeding supplemental illite to Hanwoo steers on methane (CH₄) emission and rumen fermentation parameters. An in vitro ruminal fermentation technique was conducted using a commercial concentrate as substrate and illite was added at different concentrations as treatments: 0%, 0.5%, 1.0%, and 2.0% illite. Total volatile fatty acids (VFA) were different (P < 0.05) at 24 h of incubation where the highest total VFA was observed at 1.0% of illite. Conversely, lowest CH₄ production (P < 0.01) was found at 1.0% of illite. In the in vivo experiment, two diets were provided, without illite and with addition of 1% illite. An automated head chamber (GreenFeed) system was used to measure enteric CH₄ production. Cattle received illite supplemented feed increased (P < 0.05) total VFA concentrations in the rumen compared with those fed control. Feeding illite numerically decreased CH₄ production (g/day) and yield (g/kg dry matter intake). Rumen microbial population analysis indicated that the population of total bacteria, protozoa and methanogens were lower (P < 0.05) for illite compared with the control. Accordingly, overall results suggested that feeding a diet supplemented with 1% illite can have positive effects on feed fermentation in the rumen and enteric CH₄ mitigation in beef cattle.     

Questionnaire‐based Analysis of Owner‐reported Scratching and Pain Signs in Cavalier King Charles Spaniels Screened for Chiari‐like Malformation and Syringomyelia

Background

Chiari‐like malformation (CM) and syringomyelia (SM) cause a pain syndrome in Cavalier King Charles spaniels (CKCS). Clinical signs are not consistently apparent on neurologic examination, and owner reporting of signs provides vital clinical history. However, owner questionnaires for this disease are not well developed.

Objectives

To develop a tool to capture owner‐reported clinical signs for use in clinical trials and to compare owner‐reported signs with the presence of pain on neurologic examination and SM on magnetic resonance imaging (MRI).

Animals

Fifty client‐owned CKCS.

Methods

Owners completed a questionnaire and pain/scratch map. Each dog underwent a neurologic examination and craniocervical magnetic resonance imaging (MRI). Questionnaire responses were developed into scores, area of shading for pain/scratch maps was measured, and consistency of responses between these tools was assessed. Owner‐reported findings were compared with neurologic examination findings and presence and severity of SM on MRI.

Results

Thirty‐three dogs were symptomatic and 17 asymptomatic; 30 had SM. The most common sign of pain was crying out when lifted (n = 11). Extent of shaded areas on maps positively correlated with questionnaire scores for pain (r² = 0.213, P = 0.006) and scratch (r² = 0.104, P = 0.089). Owner‐reported findings were not significantly associated with presence or severity of SM or neurologic examination findings. Owner‐reported lateralization of signs was significantly associated with SM lateralization (P < 0.0001).

Conclusions

The questionnaire and maps may be useful for clinical trials. Lack of association of owner‐reported signs with SM highlights our lack of understanding of the pathophysiology of pain in this disease.