Effect of dietary supplementation with a synbiotic (Lacto Forte) on growth performance,haematological and histological profiles,the innate immune response and resistance to bacterial disease in Oreochromis niloticus

This study investigated the effects of a synbiotic Lacto Forte on growth performance, haemato‐immunological responses, plasma bactericidal capacity, histological profiles and resistance of Oreochromis niloticus to Pseudomonas fluorescens. The experimental fish were divided into three groups; two of them (T1 and T2) were given Lacto Forte incorporated diets at increasing level (0.7 and 1.5 g/kg, respectively) and an additive‐free basal diet served as the control (T0) for 30 and 45 days. At the end of the feeding trial, a challenge was performed using a virulent strain of P. fluorescens and mortalities were recorded over an additional 14‐days period. The results showed that Lacto Forte has a pronounced effect on haematological and growth performance parameters at 1.5 g/kg rather than 0.5 g/kg (p < 0.05). Plasma lysozyme, proteases, antiproteases and bactericidal capacity were significantly enhanced (p < 0.05) in Lacto Forte‐treated groups, particularly after 45 days of supplementation. Long‐term supplementation with Lacto Forte (1.5 g/kg) induced degenerative changes in the liver, spleen and intestine. Additionally, a significant increase (p < 0.05) in survival rates were found in Lacto Forte‐treated groups compared to the control one 14 days post‐challenge. As a result, Lacto Forte fortifies tilapia immune response and can be used as a surrogate for antibiotics to control P. fluorescens.     

Pre‐challenge and post‐challenge haemato‐immunological changes in Oreochromis niloticus (Linnaeus, 1758) fed argan oil against Lactococcus garvieae

The present study investigated the effects of argan oil, obtained from Argania spinosa, on pre‐ and post‐challenge immuno‐haematological and biochemical responses of Nile tilapia, Oreochromis niloticus. For this purpose, the fish were fed diets containing 0, 0.5%, 1% or 2% argan oil for 45 days. Following 45 days of feeding, fish were challenged with Lactococcus garvieae and mortality was recorded for 15 days. During the pre‐challenge period, significantly higher respiratory burst activity, total white blood cell (WBC), serum lysozyme activity and myeloperoxidase activity were determined in the argan oil‐fed groups. The serum glucose and cholesterol levels decreased whilst total protein and albumin did not change in the groups fed with argan oil‐supplemented diets. After challenge with Lactococcus garvieae, the percentage survival (%) was found to be the highest in the 1% and 2% argan oil‐supplemented feeding groups. Also, there was a significant increase in weight gain, specific growth rate and feed conversion ratio in those fish fed argan oil. The results of this study indicated that after the supplementation of fish diets with argan oil, especially at 1% and 2% concentrations, the immunological, haematological and biochemical values remained similar in both the pre‐ and post‐challenge periods and the immune response against L. garvieae in Nile tilapia was modulated.     

Labeo rohita and Argulus siamensis infection: Host size,local inflammatory reaction and immunity modulate ectoparasite load on fish

Parasite species often show a heterogeneous, highly dispersed pattern of infestation within hosts. Varieties of factors including morphological, physiological, immunological and nutritional characteristics affect the infestation level of a specific parasite in homogenous pray. Limited attempts, however, have been made to explore such underlying drivers of infestation pattern. Here, three stages of Labeo rohita (fingerling, juvenile and pre‐adult) were challenged with ectoparasite, Argulus siamensis in same aquaria. The parasite load on individuals was determined at 5‐day interval for 1 month. The load was found to be highest in pre‐adult stage followed by juveniles and fingerlings. On day 20 post infection, the load of parasite on pre‐adult fish was high along with detectable skin damages. Skin tissues were collected for immune gene expression analysis and histopathology. Histological studies showed increased melanization in the dermis and mild inflammatory cellular reactions in pre‐adult fish whereas, massive subcutaneous myositis with engorged blood vessels were observed in fingerlings. The expression levels of various inflammation and innate immune‐related genes viz., interleukin (IL)‐8, IL‐10, IL‐11, IL‐15, natural killer enhancing factor, toll‐like receptor 4, apolipoprotein A–I and immunoglobulin Z were significantly high in skin samples of infected fingerlings as compared to other two growth stages or controls of each stage. On the other hand, the expression of immunoglobulin M was down‐regulated in all infected samples as compared to their respective controls. The results thus depict that local immuno‐inflammatory response plays a significant role in determining susceptibility of host in intra‐specific group, and has important implications for ecology and aquaculture.     

Stress response of Salmo salar (Linnaeus 1758) facing low abundance infestation of Caligus rogercresseyi (Boxshall & Bravo 2000), an object in the tank,and handling

This study looks at how low infestation loads of adult Caligus rogercresseyi and other stressors affect the physiology of Salmo salar. Experimental fish groups were with (infested) or without (control) exposure to the parasite. The parasite cohort was followed for 78 days post‐infestation (dpi), and only adult lice were observed. Additional stressors were applied at 60 and 75 dpi. The analysis included measurements of fish physiology and weight. Low‐level infestations by adult C. rogercresseyi for more than 50 dpi induced moderate stress in S. salar as well as a high energy demand and increased small skin mucous cells. Threshold lice loads were identified, and above those loads, a high stress response was observed. Additional stressors altered fish physiology, inducing downregulation of the cortisol response after the first stressor and upregulation after the second stressor, but infested fish responded more strongly. Parasitism by C. rogercresseyi is energetically demanding, affecting the primary and secondary responses (e.g. cortisol and glucose levels), as well as the tertiary response (fish weight).     

Acute effects of ammonia exposure on the plasma and haematological parameters and histological structure of the juvenile blunt snout bream,Megalobrama amblycephala,and post‐exposure recovery

In this study, we investigated the toxic effects of ammonia‐N on the plasma and haematological parameters and histological structure of blunt snout bream (Megalobrama amblycephala) juveniles. The fish (initial weight, 14.79 ± 0.01 g) were randomly sorted into six tanks (200 L), and each tank was stocked with 40 fish for culture. The juveniles were exposed to two ammonia‐N levels—0 mg/L (control group) and 25 mg/L (experimental group)—and sampled at 0, 6, 12, 24, 48 and 72 hr, and then they underwent 96 hr of post‐exposure recovery. The results showed that ammonia‐N had significant effects on the plasma and haematological parameters. The treatment group showed increased cortisol, plasma ammonia and haematocrit levels and white blood cell count with increasing exposure time, up to 24 hr, and then the levels and count decreased. A significantly higher plasma glucose level was observed in the treatment group at 12 hr. After 96 hr of post‐exposure recovery, all parameter levels decreased to the control levels. The fish displayed histopathological alterations in the gills, liver and kidney. The results indicate that the severity of the lesions clearly differed among the organs, with the liver showing the most extensive damage, followed by the gills and kidney. Adverse effects to physiological indicators and histological structure increased with increasing exposure time before 24 hr. The fish showed self‐regulation; however, the histological structure could not recover fully, the gill tissue showed irreversible changes and the kidney tissue exhibited the worst recovery ability.     

Practical diets with essential oils of plants activate the complement system and alter the intestinal morphology of Nile tilapia

The effect of the essential oils (EOs) of peppermint, Mentha piperita L., and tea tree, Melaleuca alternifolia (Maiden & Betche) Cheel, on the haematological, biochemical, and immunological parameters and intestinal morphology of Nile tilapia, Oreochromis niloticus L., was evaluated. Fish (58.09 ± 5.87 g) were fed 100 mg/kg and 250 mg/kg of each EO and sampled on days 7, 14, 30 and 60 after starting supplementation. The haematological and biochemical parameters were not altered by the supplementation of EOs compared to the control (p > .05). With regard to the immunological parameters, the activation of the complement system of fish fed 250 mg/kg peppermint and 100 mg/kg and 250 mg/kg tea tree EOs were significantly higher compared to the control after 60 days of feeding (p < .05). The complement system plays an essential role in innate immunity and contributes significantly to the acquired immune response; thus, its activation through supplementation with EOs is promising for the formulation of nutritional additives in aquaculture. Regarding intestinal morphology, fish fed 250 mg/kg tea tree EO presented higher villus size compared to all other groups (p < .05), which represents a healthier gut. These fish present a larger intestinal surface, which can result in better absorption and utilization of the nutrients. Based on the responses found in this study, both EOs were considered promising for the formulation of feed additives for Nile tilapia.     

Probiotic dietary supplementation in Nile tilapia as prophylaxis against streptococcosis

This study evaluated the effects of the probiotic Lactobacillus plantarum as dietary supplement on growth performance, haemato‐immunological responses, microbiology, histology and transmission electron microscopy of the intestinal epithelium of Nile tilapia challenged with Streptococcus agalactiae. Fish were distributed into two groups: control (unsupplemented) group and the group fed L. plantarum supplemented diet for a period of 58 days. We observed an increase in the concentration of lactic acid bacteria and a reduction in the number of Vibrionaceae in supplemented fish. A significant increase in the final weight, specific growth rate and feed efficiency was also observed in supplemented fish. After challenge, the number of thrombocytes and neutrophils also increased in supplemented animals. Transmission electron microscopy showed damage to the intestinal mucosa and the presence of bacteria similar to S. agalactiae in both infected groups. L. plantarum colonized the intestines of fish, enhanced the growth performance and modulated some haematological parameters.     

Natural co‐infection of cultured Nile tilapia Oreochromis niloticus with Aeromonas hydrophila and Gyrodactylus cichlidarum experiencing high mortality during summer

Aeromonas hydrophila and Gyrodactylus cichlidarum are common pathogens that induce significant economic losses in farm‐reared Nile tilapia. Nowadays, the sudden appearance of fish mortalities was exaggerated due to mixed and multiple infections. During summer 2016, mass mortality among earthen pond‐farmed Nile tilapia was reported. Clinico‐pathological, bacteriological and parasitological examinations have been demonstrated. As well, the water quality parameters were assessed. The clinical and histopathological findings of the moribund and recently dead fish were characterized by generalized septicaemic signs. The water quality parameters were significantly elevated over the permissible levels, whereas there was an elevation in nitrite (0.04 mg/L), un‐ionized ammonia (0.8 mg/L), hydrogen sulphide levels (153.1 mg/L) and organic matter content (3.79 mg/L). A. hydrophila was identified based on phenotypic characterization, API 20E features and the homology of 16S rRNA gene sequence analysis. In addition, PCR data confirmed the presence of aerolysin (aerA) and haemolysin (hly) genes in the identified A. hydrophila isolates. Gene sequencing and phylogenetic analysis based on 16S rRNA sequence confirmed that A. hydrophila H/A (accession No. MN726928) of the present study displayed 98%–99% identity with the 16S rRNA gene of A. hydrophila. Furthermore, the monogenetic trematode, G. cichlidarum was identified in the wet mounts from the skin and gills of the examined fish with a high infestation rate. In this context, it was reported that the synergistic co‐infection of A. hydrophila and G. cichlidarum with deteriorated water quality parameters could induce exaggerated fish mortalities during hot weather.     

Dietary addition of the essential oil from Lippia alba to Nile tilapia and its effect after inoculation with Aeromonas spp.

The objective of this study was to evaluate the effect of dietary addition of the essential oil from Lippia alba (EOLA) on growth performance, biochemical and haematological variables and survival after Aeromonas spp. infection of Nile tilapia juveniles (Oreochromis niloticus). Five diets were evaluated with increasing levels of EOLA (0.0 control; 0.25; 0.50; 1.0; and 2.0 ml/kg diet). After 45 days of feeding, the fish were infected with Aeromonas spp. followed by a 14‐day period of observation. There was no mortality during growth assessment. The addition of 2.0 ml EOLA/kg diet improved feed conversion ratio and condition factor and increased lysozyme activity and haematocrit (Hct), and decreased plasma globulin levels compared to the control group. The survival of fish infected with Aeromonas spp. was higher in those fed with 2.0 ml EOLA/kg diet than in fish fed with 0.0–0.5 ml EOLA/kg diet. We concluded that the inclusion of 2.0 ml EOLA/kg diet is recommended for Nile tilapia juveniles, because it improved feed conversion, Hct and immunological activity, did not provoke metabolic changes, and increased survival after Aeromonas spp. infection.     

Extracts of Oliviera decumbens and Satureja khuzestanica as immunostimulants affect some innate immunity indices of Cyprinus carpio against Aeromonas hydrophila infection

In this study, the effect of extracts of two herbs (Oliviera decumbens and Satureja khuzestanica) on immune response of carp was investigated. At the beginning of experiment, fish were divided to two groups including vaccinated (using vaccine developed against A. hydrophila) and non‐vaccinated. Fish in both groups were fed diets containing O. decumbens and S. khuzestanica and combination of two herbal extracts for 5 weeks. Control fish (negative control) and fish vaccinated only (positive control) were fed basal diets without supplements of herbal extracts. Lysozyme activity, antibody titre, complement activity and bactericidal activity in serum were measured. After 5 weeks feeding, fish were infected with A. hydrophila and mortalities were recorded. In both experimental groups, no significant differences were found in terms of alternative haemolytic complement (ACH50) activity and antibody titres of Serum. In non‐vaccinated fish group, lysozyme and bactericidal activity of fish fed S. khuzestanica or combination of O. decumbens and S. khuzestanica was higher compared with control and other experimental treatments. In vaccinated fish group, the lysozyme and bactericidal activity was not significant in all treatments compared with control group. The results of this study showed that feeding non‐vaccinated and vaccinated carp with only S. khuzestanica or in combination with O. decumbens enhance only some immunity indices including lysozyme and bactericidal activity in non‐vaccinated fish and these extracts have no immunological stimulatory role on vaccinated individuals.