The development and growth of fish farming are hindered by viral and bacterial infectious diseases, which necessitate effective disease control measures. Furunculosis, primarily caused by Aeromonas salmonicida, stands out as a significant bacterial disease affecting salmonid fish farms, particularly rainbow trout. Vaccination has emerged as a crucial tool in combating this disease. The objective of this experiment was to assess and compare the efficacy and duration of different vaccine protocols against furunculosis in large trout under controlled rearing conditions, utilizing single and booster administrations via intraperitoneal, oral, and immersion routes. Among the various vaccination protocols tested, only those involving intraperitoneal injection, administered at least once, proved truly effective in preventing the expression of clinical signs of furunculosis and reducing mortality rates. A single intraperitoneal administration provided protection for up to 2352°-days, equivalent to approximately 5 months in water at 16 °C. However, intraperitoneal vaccination may lead to reduced growth in the fish due to resultant intraperitoneal adhesions. Additionally, protocols incorporating booster doses via intraperitoneal injection demonstrated efficacy regardless of the administration route of the primary vaccination. Nevertheless, the use of booster vaccinations via the intraperitoneal route did not confer any significant advantage over a single intraperitoneal injection in terms of efficacy.

Furunculosis caused by Aeromonas salmonicida subsp salmonicida (Ass) is a medically and economically important bacterial disease in salmonid farms that requires therapeutic measures to prevent and control the disease. Evaluation of the effectiveness of traditional measures such as antibiotics or vaccines usually requires infecting fish experimentally. The objective of this study is to develop a method of infectious challenge of large (250-g) Rainbow trout by immersion close to natural infection conditions. We compare mortality, morbidity and anti -Ass antibody production of Rainbow trout following different bathing times (2, 4, 8 and 24 h) at a final bacterial concentration of 106 CFU/mL. One hundred sixty fish divided in five groups corresponding to the 4 bathing times and the non-challenged group were studied. The 24 h contact duration resulted in the infection of all fish, with a mortality rate of 53.25%. The challenged fish developed acute infection with symptoms and lesions (inappe-tance, altering of swimming behaviour, presence of boils) similar to those observed in furunculosis, and produced antibodies against the bacterium at 4 weeks after challenging, in contrast with the non-challenged group.

The present study aimed to investigate whether environmental enrichment had significant effects on rainbow trout's welfare through nature-, functions- and feeling-based approaches. Fish group behavior was analyzed during the rearing phase, as well as growth performance and health status. We assessed individual's emotional reactivity through a novel-tank test by measuring fear-related behaviors and stress-related physiological responses. Fish boldness and neophobia were then evaluated towards a novel object. We showed that more complex environments decreased aggression levels and improved growth, without impacting fish immune status. Enriched fish were also found less fearful when isolated in a novel tank and bolder when facing a novel object. We concluded that complexifying the environment through the addition of physical structures which stimulate and encourage fish to explore promotes rainbow trout's welfare in farming conditions, according to the three different welfare approaches.

This study presents the occurrence and abundance of Aeromonas antibiotic-resistant bacteria (ARB) and genes (ARGs) isolated from water, biofilm and fish in two commercial trout farms before and one week after flumequine treatment. Wild (WT) and non-wild (NWT) strains were determined for quinolones (flumequine, oxolinic acid and enrofloxacin), oxytetracycline (OXY), florfenicol (FFN), trimethoprim-sulfamethoxazole (TMP) and colistin (COL), and pMAR (presumptive multi-resistant) strains were classified. Forty-four ARGs for the mentioned antibiotics, β-lactams and multi-resistance were quantified for 211 isolates. BlaSHV-01, mexF and tetE were the dominant ARGs. A greater occurrence and abundance of tetA2, sul3, floR1, blaSHV-01 and mexF were observed for NWT compared to WT. The occurrence of pMAR and NWT Aeromonas for quinolones, OXY, FFN, TMP, COL and ARGs depended on the Aeromonas origin, antibiotic use and the presence of upstream activities. Our results revealed the impact of a flumequine treatment on Aeromonas present on a fish farm through an increase in NWT and pMAR strains. The link between fish and their environment was shown by the detection of identical ARB and ARGs in the two types of samples. There appears to be a high risk of resistance genes developing and spreading in aquatic environments.

Aims This study aimed to evaluatein vitroefficacy of essential oils (EOs) and their compounds (EOCs) alone or in combination againstAeromonas salmonicidasubsp.salmonicida,the causative agent of furunculosis in salmonid fish. Methods and Results Antimicrobial activity of 13 EOs and 16 EOCs was investigated for fourA. salmonicidasubsp. salmonicidastrains using broth microdilution. The checkerboard assay was used to evaluate a putative synergy between the most efficient EOs and EOCs against the tested strains. Cinnamon bark, oregano, clove, and thyme oils and their major compounds cinnamaldehyde, eugenol, carvacrol and thymol showed the lower minimum inhibitory concentration and minimum bactericidal concentration values. The association of cinnamaldehyde and eugenol (V/V: 30%/70%) showed a synergistic activity against three tested strains. The combinations of cinnamon with oregano, clove or thyme EOs showed a neutral or additive activity against all the tested strains. Conclusions Cinnamon, oregano, clove and thyme oils and their major phytochemical compounds showed strong activities againstA. salmonicidasubsp.salmonicidastrains. Significance and Impact of the Study To reduce the use of antibiotics in aquaculture, phytochemicals such as cinnamaldehyde and eugenol can be tested alone or in combination inin vivostudies as functional feed alternatives.

To limit the use of antibiotics in farmed fish and their potential negative impacts on public health and the environment, an evaluation of “functional alternatives” is required. The availability of effective treatments to control fish diseases is one of the most significant challenges facing aquacultures and veterinarians to reduce consequences of antimicrobial resistance. This paper includes results from in vivo studies in major freshwater-farmed fish species (salmonids, cyprinids, and cichlids), focusing on the efficacy of functional alternatives against Aeromonas spp. infections. It also outlines the recent biocontrol advances and potential alternative treatments in aquaculture. Functional alternative products can increase the resistance against Aeromonas spp. particularly by increasing the immunocompetence of fish. Many diverse alternative products such as probiotics, prebiotics, plants, essential oils, algae phages, minerals, and nanoparticles have been tested, but the diversity of the experimental designs makes it difficult to compare the efficacy of the tested products. It suggests the standardization of investigations on functional feed products for each fish species against a specific pathogen. This review also recommends farm research on functional feed alternatives in natural conditions in order to evaluate the decrease of antibiotic consumption in fish farms.

Yersina ruckeri is an enterobacteria responsible for Enteric redmouth disease (ERM), which causes significant economic losses in the aquaculture industry worldwide. Two biotypes have been described within Y. ruckeri: biotype 1 (BT1) and biotype 2 (BT2). Unlike BT1, BT2 is negative for motility and lipase secretion. The emergence of BT2 Y. ruckeri has been associated with disease outbreaks in vaccinated fish in several countries, notably France in the early 2000s. In this study, 15 BT2 strains (14 BT2 strains isolated in France and the BT2 reference strain EX5) were studied to compare the phenotypic characters of the BT1 and BT2 strains and to determine the genetic origin of the emergence of BT2 in France. BT1 bacteria are significantly longer in size than BT2 bacteria (a difference of 0.222 µm). The loss of motility of some French BT2 strains could be due to the loss of their ability to produce flagella caused by three mutations within the fliG, flhC and flgA genes. In the light of these results, the emergence of BT2 Yersinia ruckeri in France is discussed.

Interleukins have been well described in mice and humans. In large domestic animals the situation is drastically different and there is still a need for further researches aiming at identifying all the homologous interleukins and comparing their functions among species. We performed here a bibliometric analysis of all interleukins described in the literature in various large animal species to identify what is known so far and to underline where there is a need for new studies. Using indicators such as H index but also M quotient, A index, G index, GH ratio, and HG index we ranked 39 interleukins identified so far in bovine, caprine, equine, ovine, and porcine, the main large domestic animals. Indexes and ratio under investigations were higher for IL1, IL2, IL4, IL5, IL6, IL8, IL10, IL12, and IL18 than for other interleukins, particularly in bovine and porcine species and to a certain extent in equine species. Recently discovered interleukins presented low values for the different indexes, quotient, and ratio. Even some “old” interleukins showed low values highlighting the need for further developments in comparative immunology. For instance an interleukin such as IL4 demonstrated variation in its functions between species. In conclusion, this study provides the first bibliometric analysis dedicated to large domestic animal interleukins and underlines the need for more studies to fully determine the structure and the functions of interleukins in other mammal species.

Babesia sp. BQ1 (Lintan) is one of the parasites isolated from infected sheep in China that belongs to the B. motasi-like phylogenetic group. The rhoptry-associated-protein 1 (rap-1) locus in this group consists of a complex organization of 12 genes of three main types: 6 rap-1a variants intercalated with 5 identical copies of rap-1b and a single 3' ending rap-1c gene. In the present study, transcription analysis performed by standard RT-PCR demonstrated that the three different rap-1 gene types and the four rap-1a variants were transcribed by the parasite cultivated in vitro. Peptides, specific for each rap-1 type gene, were selected in putative linear B-epitopes and used to raise polyclonal rabbit antisera. Using these sera, the same expression pattern of RAP-1 proteins was found in parasites cultivated in vitro or collected from acute infection whereas only RAP-1a67 was detectable in merozoite extracts. However, ELISA performed with recombinant RAP-1a67, RAP-1b or RAP-1c and sera from infected sheep demonstrated that RAP-1a67 is the main RAP-1 recognized during infection, even if some infected sheep also recognized RAP-1b and/or RAP-1c. (C) 2016 Elsevier B.V. All rights reserved.

Background In China, ovine babesiosis is one of the most important tick-borne haemoparasitic diseases of small ruminants. It has a significant economic impact, and several Babesia motasi -like isolates have been recently shown to be responsible for ovine babesiosis in this country. Methods Full-length and C-terminal-truncated forms of the rap-1a61-1 gene of Babesia sp. BQ1 (Lintan) were cloned into the pET-30a plasmid and subsequently expressed as His-fusion proteins. The resulting recombinant RAP-1a proteins (rRAP-1a61-1 and rRAP-1a61-1/CT) were purified and evaluated as diagnostic antigens using Western blot analysis and ELISA. The native Babesia sp. BQ1 (Lintan) RAP-1 protein was recognized using Western blots and IFAT by antibodies that were raised in rabbits against rRAP-1a61-1/CT. The specificity, sensitivity and positive threshold values for rRAP-1a61-1/CT in ELISA were evaluated. Results Cross-reactivity was observed between rRAP-1a61-1/CT and positive sera for Babesia sp. BQ1 (Lintan), Babesia sp. BQ1 (Ningxian) and Babesia sp. Tianzhu isolates obtained from infected sheep. At one week post-inoculation, a significant increase was observed in the amount of antibodies produced against RAP-1a, and high levels of antibodies against RAP-1a were observed for 3 months (at 84 days p.i.). A total of 3198 serum samples were collected from small ruminants in 54 different regions in 23 provinces of China. These samples were tested using ELISA based on the rRAP-1a61-1/CT protein. The results indicated that the average positive rate was 36.02 %. Conclusions The present study suggests that rRAP-1a61-1/CT might be a potential diagnostic antigen for detecting several isolates of B. motasi -like parasites infection.

In Europe, most clinical babesiosis cases in humans have been attributed to Babesia divergens and Babesia sp. EU1. Babesia microti infection of humans occurs mainly in the United States; although a case of autochthonous B. microti infection and serological evidence of infection have been reported in Europe. The Indirect Fluorescent Antibody Test was used to screen sera from 199 anonymous Belgian patients with history of tick bite and clinical symptoms compatible with a tick-borne disease. The serological screen detected positive reactivity in 9% (n = 18), 33.2% (n = 66), and 39.7% (n = 79) of the samples against B. microti, B. divergens, and Babesia sp. EU1, respectively. Thus, evidence of contact among three potentially zoonotic species of Babesia and humans has been confirmed in Belgium. Preventive action and development of better diagnostic tools should help in prevention of clinical cases and to clarify the true burden of such infection for individuals and public health.

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Babesiosis is an important veterinary and zoonotic tick borne disease caused by the hemoprotozoan Babesia spp. which infects red blood cell of its vertebrate host. In order to control the infection, vaccination that targets molecules involved in the invasion process of red blood cells could provide a good alternative to chemotherapy. Among these molecules, Apical Membrane Antigen-1 (AMA-1) has been described as an excellent vaccine candidate in Plasmodium spp. In this paper, we have investigated AMA-1 of Babesia divergens (BdAMA-1) as vaccine candidate by evaluating its polymorphism and by studying the humoral response against BdAMA-1 of sheep experimentally infected with B. divergens. Polymorphism of BdAMA-1 was investigated by sequencing the corresponding gene of 9 B. divergens isolates from different geographical areas in France. Two Bdama-1 haplotypes (A and B) could be defined based on 2 non-synonymous point mutations. In silico prediction of linear epitopes revealed that the antigenicity of the 2 haplotypes is very similar. Antibody production against the extracellular domain of BdAMA-1 is weak and late, between 1 and 5 months after the inoculation of parasites. Both IgG1 and IgG2 are components of the anti-BdAMA-1 response. These results indicate that while BdAMA-1 may not be an immuno-dominant antigen, it could induce a mixed type 1 and type 2 immune response. In light of these results, the potential of BdAMA-1 as vaccine candidate is discussed.

A new gene of Babesia sp. BQ1 (Lintan) (BQP35) was cloned by screening a merozoite cDNA expression library with infected sheep serum and using rapid amplification of cDNA ends (RACE). The nucleotide sequence of the cDNA was 1140bp with an open reading frame (ORF) of 936bp encoding a 35-kDa predicted polypeptide with 311 amino acid residues. Comparison of BQP35 cDNA and genomic DNA sequences showed that BQP35 does not possess an intron. Recombinant BQP35 (rBQP35), expressed in a prokaryotic expression system, showed abnormally slow migration on SDS-PAGE. Gel shifting, amino acid sequence and in silico disorder region prediction indicated that BQP35 protein has characteristics of intrinsically unstructured proteins (IUPs). This is the first description of such proteins in the Babesia genus. BQP35 induced antibodies production as early as one week after Babesia sp. BQ1 (Lintan) infection in sheep. No cross-reaction was observed with sera from sheep infected with other ovine piroplasms dominant in China, except with Babesia sp. Tianzhu. The interest of BQP35 as a diagnostic antigen is discussed.