Immune profiling of rainbow trout (Oncorhynchus mykiss) exposed to Lactococcus garvieae: Evidence in asymptomatic versus symptomatic or vaccinated fish

Lactococcosis, caused by the Gram-positive bacterium Lactococcus garvieae, is a major concern in rainbow trout (Oncorhynchus mykiss) farms, which are regularly affected by outbreaks especially during the summer/fall months. In these farms, unvaccinated healthy and symptomatic fish can coexist with vaccinated fish. In the present study, innate (leukogram, serum lysozyme activity, peroxidase activity, antiprotease activity, bactericidal activity, total IgM and total proteins), and specific immune parameters (serum antibodies to L. garvieae) were assessed in unvaccinated adult rainbow trout naturally exposed to the pathogen, with or without evidence of clinical signs, or subjected to vaccination. Blood was drawn from all three groups, and blood smears were prepared. Bacteria were found in the blood smears of 70% of the symptomatic fish but not in any of the asymptomatic fish. Symptomatic fish showed lower blood lymphocytes and higher thrombocytes than asymptomatic fish (p ≤.05). Serum lysozyme and bactericidal activity did not vary substantially among groups; however, serum antiprotease and peroxidase activity were significantly lower in the unvaccinated symptomatic group than in the unvaccinated and vaccinated asymptomatic groups (p ≤.05). Serum total proteins and total immunoglobulin (IgM) levels in vaccinated asymptomatic rainbow trout were significantly higher than in unvaccinated asymptomatic and symptomatic groups (p ≤.05). Similarly, vaccinated asymptomatic fish produced more specific IgM against L. garvieae than unvaccinated asymptomatic and symptomatic fish (p ≤.05). This preliminary study provides basic knowledge on the immunological relationship occurring between the rainbow trout and L. garvieae, potentially predicting health outcomes. The approach we proposed could facilitate infield diagnostics, and several non-specific immunological markers could serve as reliable indicators of the trout's innate ability to fight infection

The association of Lactococcus petauri with lactococcosis is older than expected

Lactococcosis is a globally prevalent infectious disease that has a significant economic and sanitary impact on the rainbow trout industry. Lactococcus garvieae has traditionally been considered the only species implicated in the etiology of this disease, but Lactococcus petauri, a new species, has recently been implicated as another etiological agent. Both species cannot be distinguished by routine methods commonly used in diagnostic laboratories, resulting in their misidentification. In the present study, the identification of 48 isolates initially identified as L. garvieae was studied by determining their in-silico DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) values using pairwise comparisons of their whole genome sequences and the genomes of the type strains of L. garvieae and L. petauri. The genome sequences of 37 isolates from countries in which lactococcosis can be considered endemic (Spain, Italy, Türkiye, and Greece) were obtained in this study, and the genomes of 11 isolates were retrieved from the GenBank database. Isolates from Italy, Singapore, Japan, South Korea, India, one Turkish isolate from 2013 and two Spanish isolates recovered in 1992 and 1996 were confirmed as L. garvieae. The remaining isolates from Spain and Türkiye, as well as those from Portugal, Israel, USA, and Greece were identified as L. petauri. Overall, 60.4% of isolates previously identified as L. garvieae were found to be L. petauri. These results confirm the implication of both species in the etiology of lactococcosis and suggest that L. petauri plays a significant role in the epidemiology of this disease. Some of the isolates identified as L. petauri in the present study were isolated three decades ago, indicating that its association with lactococcosis is older than might be expected from the recent descriptions. The commercial Rapid ID32 Strep system was unable to discriminate between L. garvieae and L. petauri. However, both species exhibited some biochemical differences that might serve as phenotypic markers for their presumptive recognition. Consequently, isolates that hydrolyze hippurate and produce acid from sucrose and tagatose could be presumptively recognized as L. petauri, while those that fail these tests could be identified as L. garvieae. The results of this work indicate that great attention should be given to L. petauri in the epidemiology of lactococcosis

16S-23S rRNA Internal Transcribed Spacer Region (ITS) Sequencing: A Potential Molecular Diagnostic Tool for Differentiating Lactococcus garvieae and Lactococcus petauri

Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically significant infectious disease affecting farmed rainbow trout. L. garvieae had been considered the only cause of lactococcosis for a long time; however, L. petauri, another species of the genus Lactococcus, has lately been linked to the same disease. The genomes and biochemical profiles of L. petauri and L. garvieae have a high degree of similarity. Traditional diagnostic tests currently available cannot distinguish between these two species. The aim of this study was to use the transcribed spacer (ITS) region between 16S rRNA and 23S rRNA as a potential useful molecular target to differentiate L. garvieae from L. petauri, saving time and money compared to genomics methods currently used as diagnostic tools for accurate discrimination between these two species. The ITS region of 82 strains was amplified and sequenced. The amplified fragments varied in size from 500 to 550 bp. Based on the sequence, seven SNPs were identified that separate L. garvieae from L. petauri. The 16S-23S rRNA ITS region has enough resolution to distinguish between closely related L. garvieae and L. petauri and it can be used as a diagnostic marker to quickly identify the pathogens in a lactococcosis outbreak

The effects of a 6-week strength training on critical velocity, anaerobic running distance, 30-m sprint and yo-yo intermittent running test performances in male soccer players

The objectives of this study were to examine the effects of a moderate intensity strength training on changes in critical velocity (CV), anaerobic running distance (D'), sprint performance and Yo-Yo intermittent running test (Yo-Yo IR1) performances. Methods: two recreational soccer teams were divided in a soccer training only group (SO; n = 13) and a strength and soccer training group (ST; n = 13). Both groups were tested for values of CV, D', Yo-Yo IR1 distance and 30-m sprint time on two separate occasions (pre and post intervention). The ST group performed a concurrent 6-week upper and lower body strength and soccer training, whilst the SO group performed a soccer only training. Results: after the re-test of all variables, the ST demonstrated significant improvements for both, YoYo IR1 distance (p = 0.002) and CV values (p<0.001) with no significant changes in the SO group. 30-m sprint performance were slightly improved in the ST group with significantly decreased performance times identified in the SO group (p<0.001). Values for D' were slightly reduced in both groups (ST -44.5 m, 95% CI = -90.6 to 1.6; SO -42.6 m, 95% CI = -88.7 to 3.5). Conclusions: combining a 6-week moderate strength training with soccer training significantly improves CV, Yo-Yo IR1 whilst moderately improving 30-m sprint performances in non-previously resistance trained male soccer players. Critical Velocity can be recommended to coaches as an additional valid testing tool in soccer

Implementation of hazard analysis critical control point (HACCP) to a production line of beet sugar, molasses and pulp: A case study

White sugar, molasses and pulp constitute three products which are important for human and animal health. Sugar is either directly consumed or is used as the main component for confectionery purposes. Molasses and pulp are most frequently used as animal feeds and eventually contained contaminations find their way to humans when the latter consume meat. Therefore, it is of great importance to ensure the standard quality and safe production of sugar and its coproducts. Implementation of Hazard Analysis Critical Control Point (HACCP) constitutes a crucial step in this direction. Identification of critical control points made possible the control of all parameters which could eventually result in deterioration of the final product. Application of the HACCP system shows that this production line can be adequately controlled, especially when HACCP is implemented within the framework of a quality control system such as ISO 9001/2