Differences In Lysozyme Activity Research Articles

Protective Efficacy of Novel Oral Biofilm Vaccines against Photobacterium damselae subsp. damselae Infection in Giant Grouper, Epinephelus lanceolatus.

Photobacterium damselae subsp. damselae is a pathogen that mainly infects a variety of fish species. There are many antibiotic-resistant strains of Photobacterium damselae subsp. damselae. In a previously published article, we described the production method for a novel oral biofilm vaccine. In the study reported herein, we confirmed the protective effect of the oral biofilm vaccine against Photobacterium damselae subsp. damselae. Twenty-eight days after vaccination, phagocytosis increased by 256% relative to the control group. The mean albumin–globulin ratios of the vaccine groups were significantly lower than the mean albumin–globulin ratios of the control group. There were no significant intergroup differences in lysozyme activity. Mean IgM titers were significantly higher in the vaccine group than in the control group. There was a significant upregulation of the TLR 3, IL-1β, and IL-8 genes in the spleen 28 days after vaccination. The cumulative mortality of the control fish was 84% after challenging fish with the Photobacterium damselae subsp. damselae, while the cumulative mortality of the oral biofilm vaccine (PBV) group was 32%, which was significantly higher than those of the whole-cell vaccine (PWV) and chitosan particle (CP) groups. There is minimal published research on the prevention and treatment of Photobacterium damselae subsp. damselae infection; therefore, this oral biofilm vaccine may represent a new method to fill this gap.

The Potential Advantage of Myo‐Inositol as a Dietary Supplement for Juvenile Hybrid Grouper (Brown‐Marbled Grouper Epinephelus fuscoguttatus ♀ × Giant Grouper Epinephelus lanceolatus ♂)

AbstractAn 8‐week feeding trial was conducted to estimate the optimum dietary myo‐inositol (MI) requirement for juvenile hybrid grouper (Brown‐marbled Grouper Epinephelus fuscoguttatus ♀ × Giant Grouper E. lanceolatus ♂). Myo‐inositol was supplemented at 0 (control group), 150, 300, 600, 900, 1,200, and 2,000 mg/kg of diet. Basic diet contained no MI. Each diet was randomly fed to quadruplicate groups of juveniles, and each tank was stocked with 30 fish (initial weight = 7.40 ± 0.01 g [mean ± SD]). Results showed that the addition level of MI exerted no significant effect on the survival rate, feed conversion ratio, or protein efficiency ratio of juvenile grouper (P > 0.05). However, the weight gain rate in the 150‐mg/kg group was significantly higher than that in the control group (P < 0.05). The total cholesterol levels in the serum of the control and 150‐mg/kg groups were significantly higher than those of the other groups, and the triglyceride level in the 150‐mg/kg group was significantly lower than those in the other groups. The glutamine aminotransferase level was significantly higher in the control group than in the supplemented groups, but no significant difference in lysozyme activity was observed. In the liver, superoxide dismutase activity in the control group was significantly lower than that in the 150–900‐mg/kg groups, and the total antioxidant capacity in the 150‐mg/kg group was significantly higher than those in the other groups. Broken‐line regression analysis showed that the MI required for juvenile hybrid grouper was 181.47 mg/kg diet based on weight gain rate, which can significantly improve growth performance and increase the activity of serum and liver enzymes.

Dietary replacement of fishmeal with commercial protein blends designed for aquafeeds in hybrid striped bass (Morone chrysops ♂× Morone saxatilis ♀): Digestibility, growth, body composition, and nutrient retention

Four commercial protein blends—Elite 60™, Elite 65™, Elite 70™, and Pro-Cision™ (HJ Baker & Bros., Inc., Tuscola, Texas, USA)—were evaluated in hybrid striped bass (HSB) for nutrient digestibility and fishmeal (FM) replacement value. A control diet was formulated like a commercial HSB feed containing 17% FM that met or exceeded the nutritional requirements of HSB. Commercial blends replaced FM in the control diets on an ideal protein basis by supplementing the first-three limiting amino acids (methionine, lysine, threonine). Diets were formulated on a digestible protein basis (38% digestible protein), isolipidic (16%), and contained 10% lipid from marine fish sources. Fish fed the control diet generally exhibited better growth and nutrient utilization responses than the other diets. Responses to the Elite™ products were intermediate, while Pro-Cision™ lagged in both digestibility and growth responses. Digestibility of gross nutrients in each of the test products did not differ significantly from those of FM, but protein was significantly more digestible in Elite 70™ compared to that of FM and Pro-Cision™. Availabilities of leucine, phenylalanine, tyrosine, and valine in Pro-Cision™ were significantly less (92.2%, 91.3%, 91.0%, and 91.5%, respectively) than those in FM (96.1%, 96.8%, 94.6%, and 94.5%, respectively). Fish fed the Pro-Cision™ diet exhibited significantly lower cumulative feed intake, average weight, and muscle ratio compared to fish fed all other diets. Elite 60™, Elite 65™, and Elite 70™ fish did not differ in intraperitoneal fat or muscle ratio compared to fish fed the control diet. Fish fed all three Elite™ diets had significantly lower average weight (52.5–57.9 g) compared to fish fed the control diet (73.2 g). There were no differences in arginine and histidine whole-body nutrient retention among dietary treatments, while other nutrient retention efficiencies did differ across treatments. Nutrient retention of the averaged essential amino acids was not significantly different in fish fed Elite 70™ compared to fish fed the control diet. There were no significant differences in lysozyme activity and mean corpuscular hemoglobin concentration among treatments. It appears that Pro-Cision™ is not an attractive alternative to FM in HSB diets considering total feed intake, growth, and nutrient and energy retentions. However, Elite 65™ and Elite 70™ may be potential FM alternatives in HSB diets despite differences in growth performance after 13-weeks.

Saccharomyces crevices and Bacillus spp. effectively enhance health tolerance of Nile tilapia under transportation stress

The effects of added probiotics on growth performance, blood chemical, and hematological responses; as well as immune-related gene expression in Nile tilapia (Oreochromis niloticus) under transportation stress were evaluated. Fish were treated over a period of 120 days in three groups: T1, the control group (no probiotic), T2, 5.6 × 108 cfu g−1 of Saccharomyces cerevisiae, and T3, 1 × 106 cfu g−1 of Bacillus spp. The fish were then exposed to the transportation stress experiment for four hours (h). The resulted revealed that there were no significant differences in survival rate during and after transportation in all treatments. However, the survival rate of the control group significant decreased (p < .05) after 24 h post transportation stress, and significantly higher rate of cortisol, glucose, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were observed in the T1 group, compared to the T2 and T3 groups. However, significantly lower malondialdehyde (MDA) was found in T3 compared to those of T1 and T2 after 24 to 72 h post-transportation. The expression of heat shock proteins (Hsp70) and tumor necrosis factor-alpha (TNF-α) levels in the control group were significantly higher (p < .05) than those of T2 and T3 groups, especially at 0 h post-transportation stress. No significant differences in lysozyme activity and hematological profiles were detected between the control (T1) and probiotic groups (T2 and T3). In conclusion, Nile tilapia reared for 120 days in aqueous probiotic showed significantly improved health tolerance under transportation stress.

Effects of different light spectra on embryo development and the performance of newly hatched turbot (Scophthalmus maximus) larvae

Light is a key environmental factor that synchronizes various life stages from embryo development to sexual maturation in fish. For turbot, light spectra have the most influence at the larval and juvenile stages. In the current study, differences in the development of embryos and the performance of newly hatched turbot larvae exposed to five different spectra: full spectrum (LDF), blue (LDB, peak at 450 nm), green (LDG, peak at 533 nm), orange (LDO, peak at 595 nm) and red (LDR, peak at 629 nm), were examined. At 62.8 h post fertilization, a higher number of embryos exposed to short-wavelengths (LDG and LDB) had developed a heartbeat in comparison with embryos exposed to other wavelengths. Larvae exposed to the green spectrum had higher malformation rates than larvae exposed to the other spectra, indicating that larvae exposed to green light may have significantly reduced survival rates. The results of non-specific immunity parameters showed that the mRNA expression levels of cathepsin D (CTSD), cathepsin F (CTSF), catalase (CAT) and metallothionein (MT) in larvae exposed to LDB were significantly higher than those exposed to other spectra, but CAT activity in larvae exposed to LDB was significantly lower than larvae exposed to the other spectra. There was no significant difference in MT activity in larvae exposed to the five different spectra. The mRNA expression level of lysozyme (LZM) in larvae exposed to LDR was significantly higher than other spectra, while there was no significant difference in LZM activity observed in larvae exposed to LDR, LDG, LDB and LDF. The difference of the enzyme activity of total superoxide dismutase (T-SOD) was not significant among larvae exposed to the five spectra. mRNA expression of the heat shock protein 70 (HSP70) was significantly higher in newly hatched larvae exposed to LDB, LDR and LDG, indicating that larvae exposed to LDB, LDG and LDR exhibited a stress response. The mRNA expression level of the insulin-like growth factor-1 (IGF-1) and growth parameters in the newly hatched larvae exposed to the different spectra were not significantly different. The results of the present study indicate that LDO and LDF should be used for embryo incubation and newly hatched larvae when rearing turbot. This study provides a theoretical basis for optimizing the incubation light environment for fertilized turbot eggs, promoting immunity and reducing stress responses in newly hatched larvae.

Dietary supplementation of dry oregano leaves increases the innate immunity and resistance of Nile tilapia against Streptococcus agalactiae infection

This study aimed to evaluate the influence of increasing concentrations of dietary dry oregano leaves (DOL), ranging from 0 to 0.15%, on different performance parameters of Nile tilapia. Hence, two separate experimental trials testing growth factors (Trial 1) and innate immunity and disease resistance against Streptococcus agalactiae infection (Trial 2) were conducted with a total of 464 tilapia juveniles and two to four replicates. As a result, dietary inclusion of DOL did not induce significant effects on growth performance, hematological parameters, and fillet composition. However, there were significant differences in lysozyme activity between groups of DOL at 0.075% and the control (0%). Moreover, the bactericidal index of the positive control and groups supplemented with DOL were significantly higher than baseline treatment groups. For disease resistance, there was a reduction of 15% in the relative risk of streptococcosis in fish juveniles after dietary inclusion of oregano at 0.15%. In conclusion, the inclusion of DOL in tilapia diets resulted in no significant gains in terms of growth; nevertheless, the immune system and resistance of tilapia against S. agalactiae infection were significantly improved.

Visceral and serum lysozyme activities in some freshwater fish (three catfish and two carps)

The lysozyme activities were investigated in three catfish: hybrid catfish [Clarias macrocephalus (Gunther 1864) x Clarias gariepinus (Burchell 1822)], Hemibagrus filamentus (Fang and Chaux 1949), and Kryptopterus apogon (Bleeker 1851) and two carps: Oreochromis niloticus (Linnaeus 1758) and Channa striata (Bloch 1793). Fish samples used in this study were from two sites of Chiang Mai province; C. macrocephalus x C. gariepinus and O. niloticus were from Ton Payom Market, Amphoe Muang. H. filamentus, K. apogon, and C. striata were from Doi Tao Lake, Amphoe Doi Tao. The serum, brain, liver, and kidney of the catfish and carps from both sites were spectrophotometrically measured at 450 nm by ELISA reader. C. macrocephalus x C. gariepinus and H. filamentus showed the highest lysozyme activity in the serum, followed by the kidney, brain, and liver, respectively, while the highest lysozyme activity was observed in the brain of K. apogon, and the least one was observed in the liver. Among the catfish, lysozyme activities of K. apogon were significantly higher than those of the other fish (P < 0.01). However, no significant difference in lysozyme activity from the three investigated visceral organs was found. In carps, O. niloticus had the highest lysozyme activity in the serum, followed by the kidney, brain, and liver, respectively. Meanwhile, the highest lysozyme activity in C. striata was recorded in the serum, followed by the liver, brain, and kidney. However, there was no significant difference in lysozyme activity from the serum and the internal organs in these two fish. In conclusion, the innate immunity arms in these five fish species encompassed the similar function pattern, although they were differently present in their organs. The data will be used as reference ranks for these species to evaluate health status when they are exposed to some pathogens or chemical contamination in streams and reservoirs.

Evaluation of the optimum dietary arachidonic acid level and its essentiality based on growth and non-specific immune responses in Japanese eel, Anguilla japonica

A feeding trial was conducted to evaluate the optimum dietary level and the essentiality of arachidonic acid (ARA, 20:4n-6) based on growth and non-specific immune responses in Japanese eel, Anguilla japonica. A basal ARA free commercial diet was used as a control, and four other diets were prepared by supplementing 0.3, 0.6, 0.9 or 1.5% ARA. The analyzed ARA concentrations of the diets were 0.00, 0.33, 0.71, 1.06 and 1.65% diet (ARA0, ARA0.33, ARA0.71, ARA1.06 and ARA1.65), respectively. Triplicate groups of fish (initial body weight, 157±2.5g) were fed one of 5 experimental diets at a ratio of 4% body weight for 12weeks. At the end of the feeding trial, final weight, weight gain, specific growth rate and feed efficiency of fish fed the ARA0.71 diet were significantly higher than those of fish fed the ARA0, ARA0.33 and ARA1.65 diets (P<0.05). The polynomial regression analysis of weight gain indicates that the optimum dietary ARA level could be 0.92% diet. Superoxide dismutase activity of fish fed the ARA1.65 diet was significantly higher than those of fish fed the ARA0 and ARA0.33 diets. However, there were no significant differences in superoxide dismutase activity among fish fed diets ARA0.71, ARA1.06 and ARA1.65. Fish fed the ARA1.06 diet showed significantly higher lysozyme activity than those of fish fed the ARA0 and ARA0.33 diets. However, there were no significant differences in lysozyme activity among fish fed the ARA0.71, ARA1.06 and ARA1.65 diets. Therefore, the optimum dietary ARA level could be greater than 0.71% but less than 0.92%. Also, these results may indicate that dietary ARA could be one of the essential fatty acids in Japanese eel.Statement of relevanceThe findings showed the essentiality of an appropriate arachidonic acid supplementation level for optimum growth of Japanese eel.

Changes in Atlantic salmon Salmo salar mucus components following short‐ and long‐term handling stress

This study examined changes in Atlantic salmon Salmo salar epidermal mucus proteins following short- and long-term handling stress. Short-term stress consisted of a single removal of fish from water for 15 s with long-term stress consisting of daily removal of fish from water for 15 s over 21 days. In the long-term handling stress study, there was a high level of individual variability with respect to mucus alkaline phosphatase, cathepsin B and lysozyme activities, with no correlation to treatment group. There was limited or no positive correlation between lysozyme, cathepsin B or alkaline phosphatase activities and plasma cortisol. There was a significant difference in lysozyme activity for both control and stressed fish at day 21 compared to other sampling days. In the short-term study, there was again high variability in mucus enzyme activities with no difference observed between groups. Immunoblotting also showed variability in mucus actin breakdown products in both short- and long-term handling stress studies. There appeared, however, to be a shift towards a more thorough breakdown of actin at day 14 in the stressed group. This shift suggested changes in mucus proteases in response to long-term handling stress. In summary, there were correlations of some mucus enzyme/protein profiles with stress or cortisol; however, the variability in S. salar mucus enzyme levels and actin fragmentation patterns suggested other triggers for inducing changes in mucus protein composition that need to be investigated further in order to better understand the role of mucus in the response of S. salar to external stressors.

Pathogen Levels, Lysozyme, and Cortisol Response in Channel Catfish with Susceptibility Differences to Edwardsiella ictaluri

AbstractDifferences in bacterial loads, clearance rates, nonspecific immune response, and cortisol response were measured for three family groups of channel catfish Ictalurus punctatus susceptible to enteric septicemia of catfish (ESC) and three ESC‐resistant families. Resistant families had lower mortality during challenge (P &lt; 0.0001). Bacterial loads on day 5 postexposure were higher (P &lt; 0.01) in susceptible families than resistant families for blood, kidney, and spleen. Blood, kidney, and spleen bacterial levels were highly correlated. Pathogen clearance was evident by day 12 in fish from the susceptible families and measurable only in kidney and spleen from resistant families on day 20. Lysozyme activity increased on day 1 (P = 0.0074) in resistant fish and on day 2 in susceptible fish (P &lt; 0.0001). An acute stress response was evident for both resistant (21.4 ± 1.7 ng/mL [mean ± SD]) and susceptible families (30.0 ± 1.6 ng/mL). Plasma cortisol levels were elevated in susceptible fish throughout challenge, except for day 12, but in resistant fish they recovered to near basal levels immediately after an acute response on day 2. There was no correlation between cortisol and lysozyme levels. Differences in lysozyme activity and pathogen levels suggest that the nonspecific immune response was effective during the early stages of infection in resistant fish.

Observations on the inherent variability of measuring lysozyme activity in coho salmon ( Oncorhynchus kisutch)

Lysozyme activity is a common measurement of innate immunity. It has also been used to investigate genetic variation and an animal's responses to factors such as stress, infections and variations in diet. This research demonstrates the inherent variation in lysozyme activity in unstimulated coho salmon ( Oncorhynchus kisutch). The role of maternal contribution, early life stage development and fish mass are considered. Genetic variation within and between strains of coho was found to be significant at selected life stages. Our results indicate that strain differences in lysozyme activity are more accurately measured by comparing the genetic variation after the eyed stage, when maternal effects are reduced. A positive correlation between plasma/serum lysozyme activity and fish mass is reported here. In summary, this study shows the role of maternal, developmental stage and size in lysozyme activity in fish, and emphasizes the importance of considering such variables when measuring the variability of lysozyme activity in fish.

Observations on the inherent variability of measuring lysozyme activity in coho salmon (Oncorhynchus kisutch)

Lysozyme activity is a common measurement of innate immunity. It has also been used to investigate genetic variation and an animal's responses to factors such as stress, infections and variations in diet. This research demonstrates the inherent variation in lysozyme activity in unstimulated coho salmon (Oncorhynchus kisutch). The role of maternal contribution, early life stage development and fish mass are considered. Genetic variation within and between strains of coho was found to be significant at selected life stages. Our results indicate that strain differences in lysozyme activity are more accurately measured by comparing the genetic variation after the eyed stage, when maternal effects are reduced. A positive correlation between plasma/serum lysozyme activity and fish mass is reported here. In summary, this study shows the role of maternal, developmental stage and size in lysozyme activity in fish, and emphasizes the importance of considering such variables when measuring the variability of lysozyme activity in fish.

Tear lysozyme activity in frozen Schirmer strips and salivary gland biopsy as parameters of lacrimal gland function

PURPOSE: The lysozyme concentration in human tears is an important parameter for tear gland function. The decline of lysozyme in tears reflects lacrimal gland destruction. In Sjögren's patients, lacrimal gland destruction parallels labial salivary gland destruction. The objective of this study was to determine whether human tear lysozyme that was frozen on Schirmer strips at -20°C for several years maintained activity and whether there was a linear relation with inflammatory changes in labial salivary glands. METHODS: A total of 200 frozen Schirmer strips were processed. They were collected from 20 randomly selected patients each year of five consecutive years, all attending the UCSF Sjögren's Clinic. The tear lysozyme in the Schirmer strips was measured by a colorimetric assay. The average lysozyme concentration each year was calculated and compared. One third of the patients underwent labial salivary gland biopsy. The correlation was calculated between the tear lysozyme concentration and the lymphocytic focus scores in biopsy specimens. RESULTS: No significant difference of average lysozyme concentration in the Schirmer strips was found when the five different years of collection were compared. The linear relation between the tear lysozyme concentrations and the focus score in labial salivary gland biopsies showed a coefficient of r = -0.41. The linear relation between other diagnostic measurements, like Schirmer test, tear breakup time, or rose bengal staining pattern, and the focus score was lower. CONCLUSIONS: Human tear lysozyme in Schirmer strips can be stored at -20°C for at least five years. There is little difference in lysozyme activity of frozen compared to unfrozen specimens. The lysozyme concentration in tears correlates better with the lymphocytic focus score in labial salivary gland biopsy than does clinical assessment and is therefore a parameter for the actual degree of tear gland destruction.

Deamidation of lysozyme during the storage of egg white.

Deamidation of lysozyme was observed during storage in a buffer solution and in egg white. The peak corresponding to native lysozyme from Bio-Rex 70 column chromatography was gradually decreased, while the peaks corresponding to deamidated lysozyme were increased during storage in 0.1 m carbonate buffer at pH 9.5. A similar change was observed during storage in egg white, but the change in egg white was larger than that in the buffer solution. A detailed analysis of the elution peaks from the Bio-Rex 70 column suggested that one to three residues of amide in lysozyme were mainly deamidated during storage in the buffer solution, and that more than three residues in lysozyme were deamidated during storage in egg white. There were significant differences in lysozyme activity between native and deamidated lysozyme, the activity being decreased in proportion to the degree of deamidation.