Immunomodulation by dietary vitamin C in healthy and aflatoxin B1-induced immunocompromised rohu (Labeo rohita)

The aim of this study was to examine the immunomodulatory effect of high levels of dietary vitamin C in healthy and immunocompromised rohu (Labeo rohita) treated with aflatoxin B1 (AFB1). Four groups of rohu were fed experimental diets containing either no vitamin C or supplemented with vitamin C at 500 ppm for 60 days. On the first day of feeding, one group fed the high vitamin C diet and one fed the vitamin C deficient diet, were injected intraperitoneally with a single doses of AFB1 at 1.25 mg kg(-1) body weight. The effect of AFB1 and high dietary vitamin C on specific and non-specific immunity, and disease resistance against Aeromonas hydrophila were examined in the rohu. The ability of vitamin C to counteract immunosuppression induced by AFB1 was also examined. Specific immunity indicated by haemagglutination and haemolysin titres against sheep red blood cells (SRBC), and bacterial agglutination appeared to be unaffected by either the AFB1 treatment or the vitamin C enriched diet. A significant reduction was observed in the non-specific immunity of AFB1-treated fish, however, indicated by lowered bactericidal and lysozyme activities. High dietary vitamin C, on the other hand, enhanced the non-specific immunity of fish, including an enhanced phagocytic ratio and increased serum lysozyme activity. Feeding a high level of dietary vitamin C to AFB1-treated fish increased these parameters to levels similar to those found in control fish. High dietary vitamin C significantly (p < 0.05) enhanced protection against Aeromonas hydrophila infection in both healthy and immunocompromised fish. Results from this study help to establish the beneficial effect of dietary vitamin C on AFB1-induced immunosuppression, as well as confirming the immunostimulatory effect of vitamin C in rohu.     

尿素水平对青山羊血液生化指标的影响

应用饲养试验与氮平衡试验,研究了不同尿素水平的日粮对青山羊血清总蛋白(STP)、血清清蛋白(ALB)、血清球蛋白(GLB)、血清清球蛋白比值(A/G)、血清尿素氮(SUN)及血清谷丙转氨酶的含量变化的影响。试验采用4×4拉丁方试验设计,试验分4个阶段,在基础日粮一致的情况下,采用4、9、14、0(对照)g/kg·DM4种不同的处理,试验结果表明:不同尿素水平极显著影响ALB、SUN浓度和GPT活性浓度(P<0.01),对血清清球比(A/G)无显著差异(P>0.05),尿素水平与STP、GLB浓度存在负相关;血清含氮生化指标STP、ALB、GLB、A/G、GPT、SUN随时间呈动态变化,出现高峰的时间比瘤胃氨浓度出现的时间平均推迟1～2h。进食尿素后,血液生化指标由平稳逐渐缓慢升高到高峰,随后又下降并趋于平衡。     

Serum protein reference values in foals during the first year of life: comparison of chemical and electrophoretic methods

Normal reference values for serum proteins of foals from birth to 1 year of age have been established. Chemical and electrophoretic/refractometric methods for total protein, albumin, total globulin and Albumin/Globulin (A/G ratio) have also been compared. The biuret total protein method and Bromcresol Green (BCG) method on the Technicon SMA 12/60 autoanalyzer were used and compared with total protein determined via refractometry and albumin determined by Cellulose Acetate (CA) electrophoresis/densitometry. Globulin and A/G ratios were calculated from the chemical method data and compared with that obtained electrophoretically. Total protein, albumin, total globulins and A/G ratios all were in agreement at all sample times studied. Data on the subfractionation of serum globulins via CA electrophoresis is also presented. Wide variations in alpha and beta globulin levels were noted among the foal sera early in life. As a result, two distinct populations of foals with respect to both globulin content and A/G ratio were identified. One of these populations (Group A) appeared to have obtained passive immunity more slowly than the other (Group B) animals. Comparison of these data with clinical cases of foals in which failure of passive transfer was a part suggests that the A/G ratio may be useful in assessing adequate colostral antibody levels in the newborn foal.     

Dietary bovine lactoferrin induces changes in immunity level and disease resistance in Asian catfish Clarias batrachus

The effects of including bovine lactoferrin (Lf) in the diet of the Asian catfish (Clarias batrachus) on specific and non-specific immunity as well as disease resistance were investigated. The catfish were fed four different diets for 2 weeks: a commercial diet as control and the same diet supplemented with 50, 100 and 200mg bovine Lf/kg feed. After 1 and 2 weeks, serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity; natural serum haemolysin titre, lysozyme activity and oxidative radical production by neutrophils as a measure of non-specific immunity as well as disease resistance against A. hydrophila challenge to vaccinated and non-vaccinated animals were evaluated. The results showed that Lf supplements, particularly at 100mg level, significantly (P<0.05) enhanced serum lysozyme level, oxidative radical production and level of protection against A. hydrophila challenge in non-vaccinated animals irrespective of length of exposure. The specific immunity was not influenced by Lf feeding as evidenced from the bacterial agglutination titre and level of protection in vaccinated animals. As Lf feeding at 100mg/kg for 1 week is able to enhance the non-specific immunity and disease resistance of catfish efficiently, these results support the possible use of Lf as an immunostimulant for farmed catfish.     

Effect of pesticide water pollution on some haematological, biochemical and immunological parameters in Tilapia nilotica fish

Nine groups of Tilapia nilotica fish each consisting of 50 fish were used to assess the effect of pesticides on the haematological, biochemical and immunological parameters in fish. Four groups were injected with 0.05 ml of Staphylococcus aureus antigen plus complete Freund's adjuvant, the first group (G1) was exposed to lindane treatment, the second (G2) was exposed to dialdrin, the third (G3) was exposed to diazinon and the forth group (G4) was exposed to malathion. Four other groups were not immunized, but injected with the adjuvant and exposed to the pesticides, the fifth group (G5) was exposed to lindane, the sixth (G6) was exposed to dialdrin, the seventh (G7) was exposed to diazinon and the eighth group (G8) was exposed to malathion. The ninth group (G9) was exposed to water without any pesticide treatment and served as controls. Both vaccinated and non-vaccinated groups were exposed to 1/10 LC50 of the tested pesticides for 30 days. The results revealed that the mean total RBCs, WBCs counts, PCV, Hb, MCV, MCH, MCHC values were lower in vaccinated groups than in the groups exposed to the tested pesticides. The total protein, globulin and serum enzymes ALAT; ASAT values as well as macrophage phagocytic index and antibody titer were lower in vaccinated as compared to the non-vaccinated groups.     

EVALUATION OF NEW AND CURRENTLY USED DIAGNOSTIC PROCEDURES FOR BOVINE BRUCELLOSIS

SUMMARY The indirect haemolysis test (IHLT) and the rough antigen complement-fixation test (RCFT) were compared with several conventional tests using serum samples from 9 cows known to be infected with Brucella abortus. In 7 cows all the tests except the RCFT (which was developed to detect antibodies resulting from 45/20 vaccination) became positive and remained so until the cows were autopsied 6 months after infection. In the other 2 cows the Rose Bengal test was occasionally negative and the titre of the serum agglutination test only reached 100 iu briefly. The routine complement-fixation test, the IHLT and the anti-bovine globulin test reached diagnostic levels at most of the weekly test intervals but occasional low titres occurred in them all. Only very low or negative titres were detected by the RCFT.     

A preliminary study on the serum protein response in canine babesiosis

Total serum protein, albumin, globulin, globulin fractions (alpha, beta and gamma globulins) and an acute-phase protein (alpha1-acid glycoprotein) were evaluated in dogs with naturally occurring mild (Group 1), severe (Group 2) or complicated babesiosis (Group 3). Results showed that the total serum protein, albumin, A/G ratio and alpha globulins were statistically different between Groups 1 and 2. There was no statistical difference between groups with total, beta and gamma globulins. The findings from this study suggest that dogs with mild and severe babesiosis had low total serum proteins, albumin, A/G ratio and alpha globulins; dogs with complicated babesiosis showed no typical serum protein changes or patterns; and that there was no evidence of an acute-phase response detectable on serum protein electrophoresis in any of the 3 groups. A marked acute-phase response was, however, present, as measured by the alpha1-acid glycoprotein, in all 3 groups. As this was a retrospective study, the possibility that the observed responses were due in part to concurrent disease could not be excluded.     

葡甘露聚糖对饲喂黄曲霉毒素B1日粮肉仔鸡生长性能、血清指标及器官指数的影响

本文通过测定肉仔鸡生长性能、血清指标及器官指数,探讨葡甘露聚糖对黄曲霉毒素B1(AFB1)的吸附作用。选用180只体重接近的8日龄肉仔鸡随机分成3组,分别为基础日粮组、基础日粮 0.1mg/kgAFB1组、基础日粮 0.1mg/kgAFB1 0.1%葡甘露聚糖组,试验期21d。试验结果表明:(1)基础日粮中加入0.1mg/kgAFB1,肉仔鸡的体重、日增重、日采食量显著下降,料重比升高。血清中总蛋白、白蛋白含量显著下降,谷草转氨酶、谷丙转氨酶活性显著升高。肝脏、肾脏、心脏和脾脏肿胀,胸腺、法氏囊萎缩;(2)0.1mg/kgAFB1日粮中加入0.1%葡甘露聚糖,明显改善肉仔鸡的生长性能,以上相关指标基本恢复到正常水平。由此可知,葡甘露聚糖减轻或基本消除了AFB对肉仔鸡生长性能、组织器官的不良影响。     

Effect of Clostridium perfringens epsilon toxin on the blood brain barrier of mice.

It was shown that Clostridium perfringens epsilon toxin has the effect of allowing the passage of 125I polyvinyl-pyrrolidone and 125I human serum albumin into mouse brain. These substances did not enter the brains of normal control mice. The passage of albumin into the brains of mice poisoned with epsilon toxin was extremely rapid. When large doses of toxin (+/-4 000 MLD) were given death ensued within 2-3 min at which stage 1,5% of the injected albumin had already entered the brain. In cases where smaller doses were given and the time interval between injection and death was longer the figure was increased to 2-2 1/2% of the injected plasma albumin.     

Immunoglobulins in piglets from sows heat-stressed prepartum

Sows were subjected to moderate heat stress in a chamber (32 C) from d 100 of pregnancy until less than 8 h before delivery of first piglet, while control sows were in a thermoneutral chamber (21 C) or farrowing house (22 C). Blood serum and colostrum at parturition of heat-stressed sows and their piglets' serum at birth had elevated cortisol concentrations. Total protein, globulin and immunoglobulin G (IgG) concentrations in sow serum tended to decrease as parturition time was approached; albumin did not change. Total protein and IgG concentrations in colostrum at parturition and in milk 24 and 48 h later tended to be lower in heat-stressed sows. Concentrations of these four protein fractions (total, globulin, IgG and albumin) in piglet serum at birth did not differ among treatment groups, but soon after colostrum ingestion they increased markedly in all groups. Therefore, in all groups total protein remained constant while globulin and IgG decreased. Globulin concentration on d 1 was lowest in piglets from heat-stressed sows, but its rate of decrease after d 1 was not affected by sow treatment. Immunoglobulin G concentration was 11 mg/ml lower, but its rate of decrease through postnatal d 20 was slower in piglets from heat-stressed sows than in those from control sows; a 10-mg/ml difference in IgG concentration on postnatal d 1 has been associated with increased preweaning mortality in piglets. Higher cortisol concentration in serum and lower IgG in colostrum of sows under heat stress was associated in their piglets with higher serum cortisol at birth and lower serum IgG for the first 20 d postnatum.     

中药组方对黄曲霉毒素B1中毒雏鸭肝功能的影响

为观察黄曲霉毒素B1(AFB1)对试验雏鸭肝功能血清指标的影响及复方中药对AFB1的颉颃效应,本试验选用7日龄健康雏鸭90只,分为3组,每组30只。Ⅰ组为空白对照组,灌胃与试验组等体积二甲基亚砜;Ⅱ组、Ⅲ组为试验组。每天分别按0.1mg/kg剂量给Ⅱ组、Ⅲ组雏鸭灌胃AFB1一次,连续投药21d,试验期间给Ⅲ组雏鸭日粮中添加2%复方中药。分别在给雏鸭投药后7、14、21d,检测雏鸭肝功能部分血清指标。结果显示,Ⅱ组、Ⅲ组雏鸭丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)与Y-谷氨酰转肽酶(GGT)活性显著高于Ⅰ组(P<0.05),而血清总蛋白(TP)与白蛋白(ALB)含量显著下降(P<0.05);与Ⅱ组比较,在日粮中添加复方中药的Ⅲ组雏鸭各项血清指标均有显著改善(P<0.05)。说明黄曲霉毒素B1导致雏鸭肝功能发生显著的变化,而复方中药能明显改善其变化。     

连续低剂量黄曲霉毒素B1对大鼠肝肾损伤作用

用含有不同质量浓度黄曲霉素B1（AFB1）（0、4、20、25、50μg／kg）的污染饲料连续饲喂各试验组大鼠60d,于7、15、30、60d静脉取血,测定各组大鼠肝功能与肾功能生化指标。每周检测大鼠体质量,试验第60天断头处死大鼠,检测肝肾组织病理学变化并计算各组大鼠的脏体系数。试验期间备组受试动物未发现明显的中毒症状和异常情况,60d内无受试动物死亡。与对照相比,不同剂量组AFB1对大鼠体质量无显著性影响;连续喂饲7d,大鼠血清AST和BuN含量显著升高（P〈0．05）;连续喂饲15d,大鼠血清AsT、BUN和ALB含量显著升高（P〈0．05）,连续喂饲30d,大鼠血清BUN含量显著升高（P〈0．05）,连续喂饲60d,大鼠血清CR和BUN含量显著升高（P〈0．05）。在各测试质量浓度条件下,大鼠虽未发现明显的中毒症状和异常情况,试验期间无动物死亡,但与空白对照相比,各剂量组AFBl对大鼠均产生了显著的肝肾组织病理学损伤,中毒程度与给毒剂量成正相关性。随着毒素暴露时间延长,大鼠肝脏对低剂量AFBl表现出一定的耐受性,而大鼠的肾脏损伤与毒素暴露时间表现出显著的积累效应。     

The immune response of a marine teleost, Pseudopleuronectes americanus, (winter flounder) to the protozoan parasite Glugea stephani

G. stephani is an intracellular cyst-forming microsporidan parasite that is found in the intestine of winter flounder (WF) Pseudopleuronectes americanus. No detectable humoral response was seen in parasitized fish or in fish injected with either spores or spore homogenate from this parasite. Quantification of total immunoglobulin (Ig) levels showed a decrease in Ig levels rather than enhancement, 21 days after intramuscular (IM) injections of spores (3 X 10(6)/ml). When a second injection of spores was administered on day 21 and tested 3 weeks later, a further decrease in total serum Ig's occurred. A decrease in total IgM levels also occurred in WF that were simultaneously injected with G. stephani and the antigens, horse red blood cells (HRBC) or formalin-killed Klebsiella pneumonia (KP). The total Ig levels of fish injected with an antigen plus spores was not as low as those injected with the parasite alone. The Ig levels, as well as antibody titers to HRBC and KP were however, lower when compared to fish injected only with the HRBC or bacteria. Disrupted spore homogenate injected into winter flounder, showed a less marked decrease in Ig levels when compared with whole spores. When a single IM injection of spores was given, followed by two weekly injections of indomethacin (a drug that inhibits prostaglandin activity), no decrease in Ig levels occurred and levels were comparable to control (saline injected) fish.     

The immune system of cyprinid fish. Oxytetracycline and the regulation of humoral immunity in carp (Cyprinus carpio)

The antibiotic oxytetracycline (oxyTC) was administered either by mixing with food or by intraperitoneal injections. In oxyTC treated animals decreased serum immunoglobulin levels were found. The primary anti-sheep red blood cell (SRBC) response was measured by enumerating plaque forming cells (PFC). It was observed that the PFC response was depressed by 80–95% in oxyTC treated animals. When an anti-SRBC serum was injected together with SRBC the immunosuppressive effect of oxyTC was absent. A secondary anti-SRBC response was not inhibited by oxyTC. On base of the results a model for antigen presentation and the interaction between macrophages, T- and B-like cells during primary and secondary responses in fish is proposed. In primary responses cellular interaction is needed to develop a proper immune response whereas after a high antigen dose challenge SRBC might behave as a T-independent antigen.     

Effect of alternate day feeding strategy of sub-optimal protein level on haemato-biochemical responses in Labeo rohita (Hamilton) juveniles

A feeding trial was conducted for 60 days to delineate the effect of alternate day feeding strategy of sub‐optimal protein level on haematological parameters, serum parameters and phagocyte respiratory burst activity (NBT) in Labeo rohita juveniles. One hundred and thirty‐five fingerlings (1.87 ± 0.01–2.26 ± 0.05 g) were distributed in triplicate groups of each treatment, and fish were fed at 5% body weight daily. Three experimental isocaloric (401.32–410.28 kcal/100 g) diets of 30%, 25%, and 20% crude protein designated as diet A, diet B, and diet C respectively, were prepared, using locally available feed ingredients. Three different feeding schedules of normal protein diet continuously (diet A‐30%), alternate feeding of 1‐day diet A followed by 1‐day diet B (1A/1B) and alternate feeding of 1‐day diet A followed by 1‐day diet C (1A/1C) were tested. The total erythrocyte count and haemoglobin content was significantly (p < 0.05) enhanced in the group T1 fed (1A/1B), and the lowest count was recorded in the group T2 fed (1A/1C). Total leucocyte counts, total serum protein, and serum globulin were higher in the group T1 fed (1A/1B) and lower in the group T2 fed (1A/1C) as compare to control. The respiratory burst activity (NBT) of blood phagocytes and serum A‐to‐G ratio was recorded significantly difference in among the treatment groups. Based on the results of the present study, it is concluded that alternate feeding of 1‐day diet A followed by 1‐day diet B (1A/1B) is equally effective and promote the immunity in Labeo rohita juveniles.     

Blood changes in carp (Cyprinus carpio) induced by ulcerative Aeromonas salmonicida infections

Only recently Aeromonas salmonicida has been recognized as a significant bacterial pathogen in ulcerative disease of cyprinid fish. Our attempts to formulate a vaccine based on bacterial surface antigens were unsuccessful in conferring reliable protection against lethal challenge. This lead us to study pathological changes in the humoral defense system during ulcerative A. salmonicida infection in carp. High numbers of opportunist pathogens such as A. hydrophila and Pseudomonas sp. were frequently recovered from the internal organs of moribund fish, in addition to A. salmonicida. These findings together with leucopenia in moribund fish suggest that pathogenesis is characterized by a state of immune suppression. In addition, fish which had sustained a sublethal infection were not protected against a subsequent lethal challenge. However, fish previously injected with a concentrated and inactivated culture supernatant showed protection. Differential blood cell counts did not differ between experimental and control groups during sublethal infection in contrast to serum proteins. Furthermore infected non-immune carp showed a progressive decrease of immunoglobulin and total serum protein levels before the day of peak mortality whereas protected carp maintained the immunoglobulin concentration despite a decrease in protein. Our observations suggest the involvement of multiple pathogenic events, affecting different parts of the humoral defense system during ulcerative A. salmonicida infection. The immunosuppressive effects can be minimized by prior vaccination with culture supernatant.     

Long‐term feeding effects of dietary protein levels on egg production,immunocompetence and plasma amino acids of laying hens in subtropical condition

The objective of this study was to determine the effect of protein levels on production performance, immune response, liver triglyceride and plasma amino acids profile of laying hens during 21–48 weeks of age. Two groups of commercial hens (Babcock B‐308) were fed with diets with different crude protein levels (14% and 18% CP). The high protein diet (18% CP) significantly increased production performance, essential amino acid intake, spleen weight (% of body weight), ND‐titre (haemagglutination‐inhibition test), serum albumin (g/dl), serum alpha‐globulin and plasma essential amino acids (except histidine) and decreased liver triglyceride (mg/g liver) in relation to the low protein diet (14% CP). In long‐term effect, therefore, this study suggested that adequate dietary condition of amino acids, particularly methionine and branch‐chain amino acids is necessary for sustaining normal immunocompetence and achieving maximum production performance.     

Serum antibody response of Indian major carp, Labeo rohita to three species of pathogenic bacteria; Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens

The immune response to mixed whole cell antigens of Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens, the common Gram negative bacterial pathogens associated with diseases of Indian major carps were evaluated for their efficacy in triggering antibody responses in rohu, Labeo rohita (Ham.). The rohu yearlings were either immunized with antigens from single bacterial strain, A. hydrophila, E. tarda and P. fluorescens or a combination of all three. An antibody response was detected at 1st week post immunization that rose significantly (p<0.05) at 4th week post immunization in all the immunized groups. The antibody level started declining after 8th week but persisted up to 10th week post immunization in all the immunized groups. Similarly, no significant difference (p>0.05) in the antibody level was found between groups immunized with single and mixed bacterial antigens. Moreover, the use of mixed bacterial antigens did not jeopardize the specific immune response to the vaccine components. Upon challenge with single pathogen, a high relative percent survival was recorded in the group immunized with mixed bacterial antigens and was comparable to those fish immunized with the single bacteria.     

Some biochemical indices in naturally occurring fascioliasis in goats

Some biochemical indices were assayed in goats naturally harbouring Fasciola gigantica infection and compared with uninfected control goats. Infected goats had significantly lower levels of serum glucose (47.6 +/- 1.8 mg dl-1) and albumin (3.1 +/- 0.1 g dl-1) and reduced albumin:globulin ratio (1.1 +/- 0.1). Total lipid (526.8 +/- 2.4 mg dl-1), serum glutamate dehydrogenase (15.3 +/- 0.9 iu litre-1) and serum alkaline phosphatase (31.6 +/- 1.9 Kind's and King's unit dl-1) were high in infected goats. No significant changes could be recorded in serum total protein, cholesterol and phospholipids.     

Expression of immune-related genes in rohu Labeo rohita (Hamilton) by experimental freshwater lice Argulus siamensis (Wilson) infection

The crustacean ectoparasite, Argulus poses one of the major threats to carp culture due to absence of any suitable control measure. The study was undertaken to determine the expression of immune-related genes in three major immunocompetent organs viz., kidney, skin and liver of rohu (Labeo rohita) during experimental freshwater lice Argulus siamensis infection. Results showed that the expression of TLR 22-like, lysozyme G and β2-microglobulin genes in kidney was significantly (P ≤ 0.05) down-regulated in lice-infected fish. On the other hand, no significant difference (P>0.05) in CXCa, lysozyme C, TNFα and complement component 3 (C3) expression was found between uninfected control and different degrees of lice infected fish. In the skin, the expression of TLR 22-like and TNFα genes were significantly up-regulated whereas that of C3 was significantly (P ≤ 0.05) down-regulated in lice-infected fish with respect to control fish. The expression of CXCa, lysozyme C and transferrin was not detected in the skin samples of fish. In the liver, the expression of CXCa, lysozyme G, β2-microglobulin and transferrin was significantly (P≤0.05) up-regulated in lice-infected fish with respect to control fish whereas expression of C3 was significantly (P ≤ 0.05) down-regulated in lice-infected fish. The expression of TLR 22-like, lysozyme C, TNFα genes was not detected in the liver samples of fish. This study indicates that majority of the genes showed down-regulation in kidney tissue whereas up-regulation in liver and skin tissues except C3 in Argulus-infected fish. We show that infection with this parasite irrespective of intensity can also result in immune gene expression changes in tissues situated away from the site of parasite attachment and feeding. The information obtained here could be useful towards understanding the susceptibility of rohu to argulosis and mechanisms involved in protection of rohu to ectoparasitic infections, which is causing immense economic losses to freshwater aquaculture sector.