Temporal Dynamics of Plasma Catecholamines, Metabolic and Immune Markers, and the Corticosterone:DHEA Ratio in Farmed Crocodiles before and after an Acute Stressor.

Commercial crocodilian farms face significant economic and livestock losses attributed to stress, which may be linked to their adopted husbandry practices. The development of appropriate and modernized husbandry guidelines, particularly those focused on stress mitigation, is impeded by the limited understanding of the crocodilian stress response. Fifteen grower Nile crocodiles were subjected to simulated acute transport stress, with blood samples collected at various intervals post-stress. Plasma levels of corticosterone (CORT), dehydroepiandrosterone (DHEA), adrenaline, and noradrenaline were determined using high-performance liquid chromatography. Glucose and lactate were measured using portable meters and the heterophil-to-lymphocyte ratio (HLR) was determined via differential leucocyte counts. Significant differences were elicited after the stressor, with acute fluctuations observed in the fast-acting catecholamines (adrenaline and noradrenaline) when compared to the baseline. Downstream effects of these catecholamines and CORT appear to be associated with a persistent increase in plasma glucose and HLR. Lactate also showed acute fluctuations over time but returned to the baseline by the final measurement. DHEA, which is used in a ratio with CORT, showed fluctuations over time with an inverted release pattern to the catecholamines. The study highlights the temporal dynamics of physiological markers under acute stress, contributing to our understanding of crocodilian stress and potentially informing improved farming practices for conservation and sustainable management.

Bacterial communities and signatures in the stomach and intestine of juvenile P enaeus (litopenaeus) vannamei shrimp affected by acute hepatopancreatic necrosis disease.

Acute hepatopancreatic necrosis (AHPND) is a severe bacterial disease affecting farmed shrimp. Although various pathogenic bacteria associated with AHPND-affected shrimp have been described, little is known about the bacterial signatures in the stomachs and intestines when the disease occurs naturally. In this study, we characterized the microbiome of P. vannamei by high-throughput sequencing (HTS). Shrimp samples were collected from a commercial farm and divided into two groups: healthy and affected by AHPND, confirmed by PCR. Stomach and intestine samples were subjected to microbiome analysis targeting the V3-V4 region of the 16S rRNA gene. PERMANOVA analysis revealed a significant disparity in the bacterial diversity between the stomach and intestine microbiomes of these two health conditions. Our results suggest that the significant abundance of Vibrio brasiliensis and V. sinaloensis in the intestines of affected shrimp plays a role in AHPND infection. This imbalance could be mitigated by the presence of Pseudoalteromonas, Gilvimarinus, and other members of the phylum Pseudomonadota such as Cellvibrionaceae, Psychromonadaceae, and Halieaceae, which showed significant abundance in healthy intestines. This study highlights the significance of the microbial community in the differentiation of specific microbial signatures in different organs of P. vannamei. These findings offer a deeper understanding of the intricate dynamics within the shrimp microbiome under these conditions, enriching our view of AHPND progression and paving the way toward future identification of probiotics tailored for more efficient management of this disease.

Microplastics contamination in commercial fish meal and feed: a major concern in the cultured organisms.

The growing scale of plastic pollution causes a devastating impact on the aquatic ecosystem. The people largely depend on animal-based food for their protein requirements. In this study, we analysed 10 different fish meal samples and 20 feed samples used in farming to understand the level of microplastic (MPs) contamination and estimate the amount of MPs ingested by farmed fish, shrimp, and chicken through feed. The abundance of MPs in fish meal samples ranges from 210 ± 98.21 to 1154 ± 235.55 items/kg. The fish meal produced from dried fish is more prone to MPs contamination than that produced from fresh fish. In the case of fish feed, MP abundances range from 50 ± 22.36 to 160 ± 36.57 items/kg in shrimp feeds, 60 ± 26.74 to 230 ± 52.32 items/kg in fish feeds and 90 ± 25.11 to 330 ± 36.12 items/kg in chicken feeds. The exposure rate of MPs is higher in the grower- and finisher-stage feeds than in the starter feed. Fiber-shaped MPs of size 100-500 µm with PE and PP polymers were predominantly found in fish meal and feed samples. EDAX analysis showed the presence of Cr, Cd, Ti, Ni, Cu, As, Al, Pb, Hg, Cd, Ti, Fe, Ca, K, and Si in fish meal samples and Ca, Na, Zn, Cu, Ni, Cl, Al, Si, S, Pb, Cd, Ti, Cr, Mg and Fe in feed samples. The possible level of exposure of microplastic particles was calculated based on MP contamination in feed, feed consumption rate, and body weight. We estimated an MP exposure level of 531-1434 items/kg feed for farmed shrimp, 234-4480 items/kg feed for fishes, and 3519-434,280 items/kg feed for chicken. This study concludes that fish meal and feed are one of the important exposure routes of MPs to the farmed animals.

Inclusion of oil from transgenic Camelina sativa in feed effectively supplies EPA and DHA to Atlantic salmon (Salmo salar) grown to market size in seawater pens.

Atlantic salmon were fed either a diet reflecting current commercial feeds with added oil supplied by a blend of fish oil and rapeseed oil (COM), or a diet formulated with oil from transgenic Camelina sativa containing 20% EPA + DHA (TCO). Salmon were grown from smolt to market size (>3 kg) in sea pens under semi-commercial conditions. There were no differences in growth, feed efficiency or survival between fish fed the TCO or COM diets at the end of the trial. Levels of EPA + DHA in flesh of salmon fed TCO were significantly higher than in fish fed COM. A 140 g fillet from TCO-fed salmon delivered 2.3 g of EPA + DHA, 67% of the weekly requirement level recommended by many health agencies, and 1.5-fold more than the 1.5 g of EPA + DHA for COM-fed fish. Oil from transgenic Camelina supported growth and improved the nutritional quality of farmed salmon in terms of increased "omega-3" supply for human consumers.

A novel p.127Val>Ile SNP in the MTNR1A gene and its relation to litter size in Thin-tailed Indonesian ewes.

Objective: The primary objective is to identify and characterize the Single Nucleotide Polymorphisms (SNPs) within the MTNR1A gene sequence in thin-tailed Indonesian ewes to assess the possible association of MTNR1A gene polymorphism with litter size trait. Methods: Forty-seven thin-tailed Indonesian sheep were selected for the study. Genotyping involved collecting blood samples, and sequencing exon 2 of the MTNR1A gene. Results: The study identified 19 novel SNPs, with 10 being non-synonymous variations, in the MTNR1A gene of Thin-tailed Indonesian ewes. One non-synonymous SNP (rs1087815963) showed a significant association with litter size, with the GC genotype exhibiting a higher average litter size than the GG genotype. The deleterious impact of p.Val127Ile SNP was predicted by various in silico tools that predicted a highly damaging effect of p.Val127Ile SNP on the structure, function, and stability of MTNR1A. Docking reactions showed a critical involvement of this locus with the binding with melatonin. Conclusion: In conclusion, the results of our study suggest that rs1087815963 has a remarkable negative impact on the MTNR1A with a putative alteration in the binding with melatonin. Therefore, it can be stated that the implementation of the novel p.Val127Ile could be a useful marker in marker-assisted selection.

The battle against the introduced pathogenic monogenean Gyrodactylus salaris in Norwegian Atlantic salmon rivers and fish farms.

The introduced salmonid ectoparasite Gyrodactylus salaris has been detected on Atlantic salmon in 53 Norwegian rivers and in 39 Norwegian fish farms. In affected rivers, the mortality of Atlantic salmon juveniles is very high, estimated to a mean of 86%. G. salaris has been considered one of the biggest threats to wild Norwegian Atlantic salmon stocks. With various measures, the authorities have reduced the potential for further spread of the parasite to new rivers and fish farms, and G. salaris has been eradicated from 43 rivers and all fish farms. Furthermore, the eradication process is almost completed in five affected rivers located at the Norwegian west coast, while preparations for the eradication in the remaining five rivers in the southeastern part of Norway have begun. The goal of Norwegian management is to eradicate the introduced pathogenic G. salaris strains from all occurrences in Norway. In fish farms, the parasite has been removed by mandatory slaughter of infected fish. In rivers, G. salaris has mostly been removed by killing all the fish hosts with rotenone. The indigenous genetic Atlantic salmon stocks are re-established after eradication of the parasite. New methods are developed using chemicals that kill the parasite without killing fish in the rivers. Norwegian authorities have so far used more than NOK 1.5 billion on research, monitoring and combating G. salaris. However, the benefits are considered many times greater than the spending. Without control measures, G. salaris would likely have spread to new Atlantic salmon rivers where the same catastrophic outcome had to be expected. The Norwegian authorities seem to meet the goal in their long-term work to halt the spread of G. salaris and to eradicate the parasite in affected rivers.

Consumer Perceptions of the Canadian Salmon Sector and Their Associations with Behaviors: A Perspective from Indigenous Rights.

Previous studies on consumer perceptions and behaviors of salmon have often neglected Indigenous rights within the Canadian salmon sector. This study innovatively addresses this gap by integrating Indigenous rights into the current analysis, alongside considerations of sustainability practices, socio-economic impacts, and consumer motivations. Our research objectives aim to fit three consumer perceptions-environmental sustainability, economic considerations, and Indigenous rights-and to evaluate their associations, alongside perception of a price increase, socio-demographics, and consumer motivation factors, with purchasing behaviors related to Canadian salmon products. Data for this study was collected from a nationwide online survey. Responses to Question 2 and Question 35 are encoded with numerical values ranging from 1 to 5, where larger numbers indicate stronger agreement with the statement. The inclusion of methodologies such as the Graded Response Model (GRM) and Cumulative Link Models (CLM) adds another innovative dimension to this study. Our findings demonstrate how consumer profiles are associated with these four perceptions and their underlying determinants. Furthermore, the study quantifies the influence of these four perceptions on each consumer purchase behavior. The implications of these findings extend to the realm of mathematical modeling in consumer decision-making processes, offering practical insights for businesses and marketers, and emphasizing the importance of implementing regulatory frameworks and initiatives that promote sustainability, safeguard Indigenous rights, and address socio-economic disparities.

Outbreak of Natural Severe Fever with Thrombocytopenia Syndrome Virus Infection in Farmed Minks, China.

We isolated severe fever with thrombocytopenia syndrome virus (SFTSV) from farmed minks in China, providing evidence of natural SFTSV infection in farmed minks. Our findings support the potential role of farmed minks in maintaining SFTSV and are helpful for the development of public health interventions to reduce human infection.

Determination of Multiclass Antibiotics in Fish Muscle Using a QuEChERS-UHPLC-MS/MS Method.

The surging global demand for fish has increased aquaculture practices, where antibiotics have become indispensable to prevent diseases. However, the passive incorporation of these compounds into the diet may have adverse effects on human health. In this work, the QuEChERS method combined with ultra-high-performance liquid chromatography tandem mass spectrometry was applied for the determination of 10 multiclass antibiotics (5 quinolones, 2 sulfonamides, 2 diaminopyrimidines, and 1 macrolide) in muscle tissue of farmed fish (European sea bass and gilt-head sea bream). The applied method demonstrated acceptable recovery values, mostly between 70 and 120%, with limits of quantification of the method meeting the established EU maximum residue limits. The analysis of twenty fish samples in duplicate revealed that most antibiotics were not present, with the only exception of oxolinic acid and tilmicosin in European sea bass, which were below the limit of quantification of the method.

Genetic analyses verify sexually mature escaped farmed Atlantic cod and farmed cod eggs in the natural environment.

Elucidating the effects of domesticated organisms escaping into the natural environment represents a topic of importance in both evolutionary and conservation biology. However, when excluding the abundant data on salmonids, there is a lack of knowledge on this topic for marine fish aquaculture, which continues to expand globally. In order to bridge this empirical gap, we investigated a suspected escape of sexually mature domesticated Atlantic cod from a commercial marine fish farm in northern Norway. This involved genotyping samples of fish from cages on the farm, putatively identified escapees and wild cod captured in the region and samples of recently spawned eggs collected in the sea. Genetic analyses confirmed a farmed ancestry of the suspected escapees, and significantly, 27% of the sampled cod eggs. Furthermore, statistical analyses revealed a strong reduction in genetic variation in all samples of the farmed cod, including low effective population size and high degree of siblingship. These results thus document the escape of sexually mature adult cod and the release of fertilized domesticated cod eggs into the natural environment. Although it is possible that some of the mature escapees spawned post-escape, the fact that only a single egg of potential hybrid farmed × wild origin was identified, together with the high number of mature cod in the farm, points to within cage spawning as the primary source of these eggs. This suggestion is supported by oceanic particle-drift modelling, verifying that transport of eggs between the farm and the egg sampling locations was plausible. This study represents a rare documentation of interaction between domesticated and wild populations for a marine fish, pointing towards potential impacts on the local wild population.

Improvement in Muscle Fatty Acid Bioavailability and Volatile Flavor in Tilapia by Dietary α-Linolenic Acid Nutrition Strategy.

To investigate the modification of muscle quality of farmed tilapia through dietary fatty acid strategies, two diets were formulated. Diet SO, using soybean oil as the lipid source, and diet BO, using blended soybean and linseed oils, each including 0.58% and 1.35% α-linolenic acid (ALA), respectively, were formulated to feed juvenile tilapia for 10 weeks. The muscular nutrition composition, positional distribution of fatty acid in triglycerides (TAGs) and phospholipids (PLs), volatile flavor, lipid mobilization and oxidation were then analyzed. The results showed that there was no distinct difference between the SO and BO groups in terms of the nutrition composition, including crude protein, crude lipid, TAGs, PLs, and amino acid. Although the fatty acid distribution characteristics in ATGs and PLs showed a similar trend in the two groups, a higher level of n-3 PUFA (polyunsaturated fatty acid) and n-3 LC-PUFA (long-chain polyunsaturated fatty acid) bound to the glycerol backbone of TAGs and PLs was detected in the BO group than the SO group, whereas the opposite was true for n-6 PUFA. Additionally, the muscular volatile aldehyde and alcohol levels were higher in the BO group. Moreover, the expression of enzymatic genes and protein activities related to lipid mobilization (LPL, LPCAT, DGAT) and oxidation (LOX and GPX) was higher in the BO group. The results demonstrate that high-ALA diets may improve the fatty acid bioavailability and volatile flavor of tilapia by improving the lipid mobilization and oxidation, which provides new ideas for the improvement of muscle quality in farmed fish.

Insights from Hi-C data regarding the Pacific salmon louse (Lepeophtheirus salmonis) sex chromosomes.

Salmon lice, Lepeophtheirus salmonis (family Caligidae), are ectoparasites that have negatively impacted the salmon aquaculture industry and vulnerable wild salmon populations. Researchers have studied salmon lice to better understand their biology to develop effective control strategies. In this study, we updated the chromosome-level reference genome assembly of the Pacific subspecies of L. salmonis using Hi-C data. The previous version placed contigs/scaffolds using an Atlantic salmon louse genetic map. By utilizing Hi-C data from Pacific salmon lice, we were able to properly assign locations to contigs/scaffolds previously unplaced or misplaced. This resulted in a more accurate genome assembly and a more comprehensive characterization of the sex chromosome unique to females (W). We found evidence that the same ZW-ZZ mechanism is common in both Atlantic and Pacific subspecies of salmon lice using PCR assays. The W chromosome was approximately 800 kb in size, which is ∼30 times smaller than the Z chromosome (24 Mb). The W chromosome contained 61 annotated genes, including 32 protein-coding genes, 27 long noncoding RNA (lncRNA) genes, and 2 pseudogenes. Among these 61 genes, 39 genes shared homology to genes found on other chromosomes, while 20 were unique to the W chromosome. Two genes of interest on the W chromosome, prohibitin-2 and kinase suppressor of ras-2, were previously identified as potential sex-linked markers in the salmon louse. However, we prioritized the 20 unique genes on the W chromosome as sex-determining candidates. This information furthers our understanding of the biology of this ectoparasite and may help in the development of more effective management strategies.

Most of the escaped farmed salmon entering a river during a 5-year period were infected with one or more viruses.

Disease interactions between farmed and wild populations have been poorly documented for most aquaculture species, in part due to the complexities to study this. Here, we tested 567 farmed Atlantic salmon escapees, captured in a Norwegian river during 2014-2018, for five viral infections that are prevalent in global salmonid aquaculture. Over 90% of the escapees were infected with one or more viruses. Overall prevalences were: 75.7% for piscine orthoreovirus (PRV-1), 43.6% for salmonid alphavirus (SAV), 31.2% for piscine myocarditis virus (PMCV), 1.2% for infectious pancreatic necrosis virus (IPNV) and 0.4% for salmon anaemia virus (ISAV). A significantly higher prevalence of PMCV infection was observed in immature compared to mature individuals. The prevalence of both SAV and PMCV infections was higher in fish determined by fatty acid profiling to be 'recent' as opposed to 'early' escapees that had been in the wild for a longer period of time. This is the first study to establish a time-series of viral infection status of escapees entering a river with a native salmon population. Our results demonstrate that farmed escapees represent a continuous source of infectious agents which could potentially be transmitted to wild fish populations.

Automated Hematological Approach and Protein Electrophoretic Pattern in Tilapia (Oreochromis niloticus): An Innovative and Experimental Model for Aquaculture.

This study aimed to assess the usefulness of two innovative automated methods (automated blood count counters and flow cytometry) for hematological investigation in Tilapia to make a contribution to the clinical diagnostics of this farmed species. Moreover, serum total proteins and their electrophoretic fractions (prealbumin, albumin, α-, ß-, and γ-fraction), as health condition indicators, were assessed. The analysis of serum total proteins and electrophoretic fraction showed a normal and typical electrophoretic pattern of healthy fish (serum total proteins, 3.70 ± 0.62 g/dL; prealbumin, 0.44 ± 0.20 g/dL; albumin, 1.17 ± 0.66 g/dL; α-fraction, 1.49 ± 0.64 g/dL; ß-fraction, 0.32 ± 0.16 g/dL; and γ-fraction, 0.29 ± 0.13 g/dL). The relationships between the values of red blood cells (RBCs), white blood cells (WBCs), and thrombocytes (TCs) obtained with the two automated methods were determined using Pearson correlation analysis. The results showed a significant positive correlation between automatic blood cell counting and flow cytometry analysis for RBCs (r = 0.97, p < 0.0001) and WBCs (r = 0.91, p < 0.0001), whereas no correlation was found for TCs (r = -0.11, p = 0.66). The preliminary results gathered in this study seem to highlight the usefulness of the new analytical techniques herein investigated in tilapia, suggesting their application in the hematological investigation of farmed fish species and their usefulness for monitoring the health and well-being of fish reared in aquaculture.

Whole genome sequencing of nine Vibrio parahaemolyticus strains encoding (Pir) toxin-like genes from shrimp cultures in northern Peru using Oxford Nanopore technology.

Nine Peruvian isolates of Vibrio parahaemolyticus were characterized through sequencing, revealing the presence of simple sequence repeat, Pir toxin-like genes, and genes associated with antibiotic resistance, toxic components, and transposable elements. These findings expand our understanding of the genetic diversity, disease resistance, and virulence in cultivated shrimp populations in Peru.

SARS-CoV-2 Outbreaks on Mink Farms-A Review of Current Knowledge on Virus Infection, Spread, Spillover, and Containment.

Many studies have been conducted to explore outbreaks of SARS-CoV-2 in farmed mink and their intra-/inter-species spread and spillover to provide data to the scientific community, protecting human and animal health. Studies report anthropozoonotic introduction, which was initially documented in April 2020 in the Netherlands, and subsequent inter-/intra-species spread of SARS-CoV-2 in farmed mink, likely due to SARS-CoV-2 host tropism capable of establishing efficient interactions with host ACE2 and the mink hosts' ability to enhance swift viral transmission due to their density, housing status, and occupational contacts. Despite the rigorous prevention and control measures adopted, transmission of the virus within and between animal species was efficient, resulting in the development of mink-associated strains able to jump back and forth among the mink hosts and other animal/human contacts. Current knowledge recognizes the mink as a highly susceptible animal host harboring the virus with or without clinical manifestations, furthering infection transmission as a hidden animal reservoir. A One Health approach is, thus, recommended in SARS-CoV-2 surveillance and monitoring on mink farms and of their susceptible contact animals to identify and better understand these potential animal hosts.

SARS-CoV-2 Infection in Beaver Farm, Mongolia, 2021.

We report an outbreak of COVID-19 in a beaver farm in Mongolia in 2021. Genomic characterization revealed a unique combination of mutations in the SARS-CoV-2 of the infected beavers. Based on these findings, increased surveillance of farmed beavers should be encouraged.

Cellulitis and bacteremia caused by the fish pathogen,Streptococcus iniae, in an immunocompromised patient: Case report and mini-review of zoonotic disease, lab identification, and antimicrobial susceptibility.

Streptococcus iniae is a fish pathogen that can also infect mammals including dolphins and humans. Its prevalence in farmed fish, particularly tilapia, provides potential for zoonotic infections, as documented by multiple case reports. Systematic clinical data beyond cellulitis for S. iniae infection in humans, including antimicrobial susceptibility data, are unfortunately rare. Here, we present a case of cellulitis progressing to bacteremia caused by Streptococcus iniae in a functionally immunocompromised patient based on CDK4/CDK6 inhibitor and endocrine therapy, and we discuss risk factors, identification, and antimicrobial susceptibility of this rare pathogen.

The distribution of strongylid egg and lungworm (Dictyocaulus eckerti) larval counts in adult female farmed red deer (Cervus elaphus), and the implications for parasite control.

Whilst healthy adult farmed red deer show little clinical indication of parasite infection, they may still be maintaining infection levels on the farm through low-level shedding of nematode eggs and lungworm larvae. This work was undertaken to establish the long-term distribution of parasite counts, to determine whether the higher counts seen in previous trials are repeatable across the same animals. All adult female red deer on a New Zealand North Island property were faecal sampled (n = 209), weighed, and body condition scored (BCS) on five sampling occasions from March - August 2021. Faecal samples were processed by modified Baermanns to recover, identify, and enumerate lungworm 1st stage larvae (FLC), and nematode faecal egg counts (FEC) were determined by mini-FLOTAC. Between animal variation for FEC was significant (p < 0.001); whilst many counts were low to zero, a few individuals were consistently shedding higher egg counts. Younger animals tended to have higher egg counts (p = 0.003), but there was no association between FEC and BCS (p = 0.22), and FEC and liveweight (p = 0.58). Modelling of the data indicated that 50% of the egg output resulted from 21% of the animals. Additionally, there was no significant association between the higher egg counts and the gastrointestinal nematode classification; 'long tails' (likely Oesophagostomum sp.) p = 0.76, and the Ostertagiinae complex p = 0.75. Lungworm counts tended to be very low (0 - 26 lpg); consistent with previous trials and literature in farmed adult deer. However, between animal differences were statistically significant (p < 0.001) indicating some animals were passing more larvae than others, and poorer conditioned animals (BCS 2.5) were significantly associated with higher larval count (p = 0.03). There was no relationship between larval count and age (p = 0.62) and larval count and liveweight (p = 0.22). Modelling indicates that 50% of pasture larval contamination was contributed by 15% of the animals. There was no correlation between nematode egg count and lungworm larvae count (p = 0.22). Adult deer may play an important role as a source of infection for young deer, therefore, an improved understanding of the distribution of infection is needed to improve parasite control.

Long-term regulation of fat sensing in rainbow trout (Oncorhynchus mykiss) fed a vegetable diet from the first feeding: focus on free fatty acid receptors and their signalling.

Taste plays a fundamental role in an animal's ability to detect nutrients and transmits key dietary information to the brain, which is crucial for its growth and survival. Providing alternative terrestrial ingredients early in feeding influences the growth of rainbow trout (RT, Oncorhynchus mykiss). Thus, the present study aimed to assess the influence, via long-term feeding (from the first feeding to 8 months), of alternative plant ingredients (V diet for vegetable diet v. C diet for a control diet) in RT on the mechanism of fat sensing at the gustatory level. After the feeding trial, we studied the pathways of the fat-sensing mechanism in tongue tissue and the integrated response in the brain. To this end, we analysed the expression pattern of free fatty acid receptors (ffar) 1 and 2, markers of calcium-signalling pathways (phospholipase Cß, Orai, Stim or Serca), the serotonin level (a key neurotransmitter in taste buds) and the expression pattern of appetite-regulating neuropeptides in the hypothalamus (central area of appetite regulation). The results revealed that the V diet modified the expression pattern of ffar1 and paralogs of ffar2 genes in tongue tissue, along with differential regulation of calcium-signalling pathways and a defect in serotonin level and brain turnover, without influencing neuropeptide expression. This study is the first to support that changes in feeding behaviour of RT fed a V diet could be due to the difference in nutrient sensing and a decrease in hedonic sensation. We revealed that RT have similar fat-detection mechanisms as mammals.