The Effect of Macronutrient Restrictions on Gut Microbiome and Biochemical Parameters of Wistar Albino Rats

Macronutrients serve as a source of energy for both gut microbiota and its host. An increase or decrease in macronutrients can either increase or decrease the composition of gut microbiota, leading to gut dysbiosis which has been implicated in many diseases state including non-communicable diseases. To achieve this, seven diets were formulated by restricting 60% of each macronutrient. These diets were fed on 42 albino rats (Wistar), divided into 7 groups of 6 rats each. Group 1 was fed on a normal laboratory chow diet (ND), group 2 received a fat-restricted diet (FRD), group 3 received a protein-restricted diet, (PFD), group 4 received a carbohydrate-restricted diet (CRD), group 5 received a protein and fat-restricted diet (PFRD), group 6 re-ceived a carbohydrate and fat-restricted diet (CFRD) and group 7 received a carbohydrate and protein-restricted diet (CPRD). Feed and water intake were given ad libitum and daily weight and food intake were recorded. The experiment went on for 4 weeks after which animals were sacrificed and intestinal content and blood were collected for analysis (gut microbial composition, glucose, insulin levels, serum lipid, and enzyme). Compared to the control group results showed a decrease in Bacteroides (40.50 - 14.00 CFU), HDL (68.20 - 40.40 mg/dl), and AST (66.62 - 64.74 U/L) in FRD. An increase in AST (66.6 - 69.43 U/L), Bifidobacterial (59.50 - 92.00 CFU) and decreased Bacteroides (40.5 - 19.5 CFU) for PRD was also recorded. CRD reduced Lactobacillus (73 - 33.5 CFU), total bacterial count (129 - 48 CFU), HDL (68.2 - 30.8 mg/dl), and cholesterol (121.44 - 88.65 mg/dl) whereas intestinal composition of E. coli (30.5 - 51.5 CFU) increased. PFRD increased Lactobacillus (73.00 - 102.5 CFU), Bifidobacterial (59.5 - 100 CFU), HDL (68.2 - 74.7 mg/dl), and Triglyceride (111.67 - 146.67 mg/dl) concentration. Meanwhile, a reduction in Bifidobacterial (59.5 - 41.5 CFU), and an increasing of AST (66.62 - 70.30 U/l) were recorded for CFRD. However, Bacteroides (40.5 69.5 CFU), LDL (30.95 - 41.98 mg/dl) increased and Bifidobacterial (59.5 - 38.00 CFU) and HDL (68.2 - 53.5 mg/dl) decreased for CPRD. This work, therefore, concludes that macronutrient restriction causes significant changes in serum marker and enzyme profile, and gut microbial composition which can cause gut dysbiosis and later on could expose the host to inflammatory diseases in the long run.

Biochemical Liver Functions and Molecular Identification of Fasciola hepatica from Experimentally Infected Rat Model

The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2nd, 4th, 6th, 8th, 10th, 12th and 14th weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14th WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.

Effects of soybean meal replacement with fermented alfalfa meal on the growth performance,serum antioxidant functions,digestive enzyme activities,and cecal microflora of geese

Fermented forages are important feedstuffs. Bacillus subtilis inoculants are often used to improve the value of forage legume fermentation. The present work was conducted to study the effects of replacing soybean meal with solid-state fermented alfalfa meal（FAM） with B. subtilis ACCC 01746 on growth performance, serum antioxidant and digestive enzyme activities, and cecal microflora in goose. 300 healthy geese with similar body weights were randomly assigned to six treatment groups with five replicates of 10 geese（five males and five females） each. Geese were fed ad libitum for 35 days. Results showed that the geese fed with 4 and 8% FAM exhibited no significant effects on their final body weights（FBW） and average day gain（ADG）（P〉0.05）, whereas 12% or higher FAM caused poor growth of the geese compared with control diet（linear（L）： P〈0.05）. The average daily feed intake（ADFI）（quadratic（Q）： P〈0.05） and feed conversion ratio（FCR）（L： P〈0.05） with 8% or higher supplementation level were higher than those of the control group. The activities of antioxidant enzyme in serum increased, and the level of malondialdehyde（MDA） significantly decreased with increasing dietary FAM level（L： P〈0.05）. However, no significant differences were observed at 8% or lower supplementation level for glutathione peroxidase（GSH-Px） and superoxide dismutase（SOD）（P〉0.05） and at 4% for catalase（CAT） supplementation level compared with the control group. All diets containing FAM increased digestive enzyme activities in geese. However, geese fed diets with 12% FAM supplementation showed the highest trypsin activities in pancreas（Q： P〈0.05）. Supplementation with 12% or higher FAM significantly increased amylase activities in pancreas（L： P〈0.05） and duodenum（L： P〈0.05） compared with the control group. Significant differences were not observed in total anaerobic bacteria between geese fed with FAM and control diets on day 35（P〉0.05）. The numbers of Bifidobacterium and Lactobacillus in the cecum of geese fed with FAM significantly increased（L： P〈0.05）, but no significant effects were found with 4 and 8% FAM supplementation levels compared with the control（P〉0.05）. By contrast, the coliform counts of cecum decreased with increasing inclusion of FAM, but these counts were significantly reduced in geese fed diets with 12% or higher FAM supplementation level（L： P〈0.05）. Collectively, our results indicated that supplementation of the basal geese diet with 8% FAM had no apparent adverse effects on growth performance, serum antioxidant enzyme activities, and digestive parameters and beneficial microbiota.

Effect of Perinatal Bisphenol A Exposure on Serum Lipids and Lipid Enzymes in Offspring Rats of Different Sex

Rats were exposed to1 or 10 μg/mL bisphenol A （BPA） in water during pregnancy and lactation. Offspring rats were given normal water and a standard diet from weaning to postnatal day （PND） 50. Perinatal exposure to BPA resulted in significantly increased body weight, visceral adipose tissue, abnormal serum lipids, and lower adiponectin （ADP） levels in both female and male offspring rats. Liver adipose triglyceride lipase （Atgl） mRNA levels and ADP protein in visceral adipose tissue were significantly decreased in BPA-exposed offspring rats. In both female or male offspring rats, obesity and dyslipidemia induced by perinatal exposure to BPA were associated with down regulation of Atgl mRNA in liver and ADP protein in visceral adipose tissue.

Interaction between Aluminum and Zinc or Copper and Its Effects on the Pituitary-Testicular Axis Ⅱ. Testicular Enzyme and Serum Gonadotropin Assay

Sprague-Dawley male rats were divided into groups and maintained on a semipurified diet containing either 5 or 40 ppm of zinc or 2 or 8 ppm of copper. Half of the rats in each group received 1000 ppm aluminum in the diet. It was found that aluminum accumulated in the pituitary glands and testes when dietary copper levels were suboptimal. The ALP activity in testes was depressed by the added aluminum when the intake of zinc or copper was suboptimal. SDH, LDH, and LDH-X activities were inhibited and GRS and GGTP activities were elevated in rats fed either the suboptimal zinc or copper diet. However, the added aluminum in the diet reversed the changes to normal levels. The testosterone levels in plasma changed very little when the zinc or copper intake was suboptimal. An increase in plasma FSH was demonstrated in groups of both suboptimal zinc and copper intake. But the plasma LH was elevated only in the group receiving the suboptimal copper diet, and the added aluminum reversed plasma LH to control levels. A lower level of testosterone was demonstrated in the group given suboptimal copper with aluminum. It was concluded that dietary aluminum influenced the pituitary-testicular axis by interacting with certain essential trace metals, particularly zinc. 1990 Academic Press. Inc.

HOMOGENEOUS ENZYME IMMUNOASSAY OF SERUM AFP BY FLUORIMETRIC KINETIC METHOD

The activity of the Ab-bound enzyme(HRP)changed after immunochemical reaction,and can be indicated by a sensitive fluorimetric kinetic method.The finding set the stage for developing sensitive homogeneous immunoassay.AFP was measured as an example.

Effects of total soy saponins on free radicals in the quadriceps femoris,serum testosterone,LDH,and BUN of exhausted rats

Purpose:The aim of this study was to investigate the impact of total soy saponins(TS) on the free radical metabolism from the quadriceps femoris muscle,serum testosterone,lactate dehydrogenase(LDH),and blood urea nitrogen(BUN) in rats exercised to exhaustion.Methods:A one-time exhausted treadmill exercise session was used.Sprague-Dawley rats were divided into 4 groups:a control group—animals receiving no TS and no exercise(NTSNE),animals receiving TS but no exercise group(TSNE),animals receiving no TS but exercised to exhaustion group(NTSE),and animals receiving TS and exercised to exhaustion group(TSE).The TSNE and TSE groups were fed TS at a dosage of 20 mg/kg body weight once per day for 2 weeks.The NTSE group was given a placebo,and the NTSNE group was not given any treatment.The NTSE and TSE groups were exercised at speed of 30 m/min on treadmill until exhausted.The exercise time and exercise distance were recorded when the rats became exhausted and the rats were then decapitated and anatomized immediately.A 10% homogenate of the quadriceps femoris tissue was prepared.The levels of superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),glutathione peroxidase(GSH-Px),glutathione reductase(GR),reduced glutathione(GSH),total antioxidant capacity(T-AOC),LDH,BUN,and serum testosterone were tested.Results:TS significantly increased the exercise time to exhaustion by 20.62%(p < 0.05).The MDA levels were decreased significantly in the TSNE group than in NTSNE group(p < 0.05);the T-AOC levels increased significantly in the TSNE group than in the other 3 groups(p < 0.01,p < 0.05,p < 0.05).The LDH activity significantly increased in the NTSE group than in TSNE group(p < 0.05).The BUN levels significantly increased in the NTSE group than in the other 3 groups(p < 0.01,p < 0.01,p < 0.05),and significantly increased in the TSE group than in NTSNE and TSNE groups(both p < 0.01).The serum testosterone levels increased significantly in the TSNE group than in the other 3 groups(all p < 0.01).SOD,CAT,GSH-Px,GR,and GSH were not statistically different among the groups.Conclusion:TS can significantly improve the exercised rats' serum testosterone level and antioxidant activity in their quadriceps femoris to varying degrees,decrease MDA and serum LDH and BUN levels,increase the exercise time,and delay the occurrence of the fatigue.

Biochemical and histopathological profiling of Wistar rat treated with Brassica napus as a supplementary feed

Metabolic changes together with cardiovascular and hepatic factors are related to the development of diseases like myocardial lipidosis,heart disease,and profound toxicity.The aim of this animal study is to determine the effects of high erucic acid containing rapeseed oil(Brassica napus L.)varieties on liver,kidney and heart muscles in Wistar rats.Male Wistar rats were divided into three groups where each group containing four rats.Group A was considered as control diet group,while Group B rapeseed wild oil group and Group C rapeseed hybrid oil group were considered as experimental diet groups.The levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),creatine kinase-MB(CK-MB)and creatinine of two experimental groups were significantly elevated while compared to the control groups(p<0.05–0.001).Nevertheless,an increment in weight retardation(p<0.05)was also observed in rapeseed hybrid oil treated groups.No significant weight retardation found in other two groups(p>0.05).Noticeable tissue injury observed in this study is a sign of the relative toxicity of erucic acid containing rapeseed oil to mammalian species.The use of Brassica napus as a supplementary feed ingredient should be,therefore,thoroughly considered.

Ensiled Cassava Peel and Caged Layers＇ Manure Mixture as Energy Source in Broiler Starter Diet

The replacement of maize with dried cassava peel and dried caged layers＇ manure mixed in ratio 5：1 （w/w）, ensiled for 14 days was investigated in broiler starter diet to evaluate their growth responses. The fermented mixture was used to partially replace maize at 0, 25% and 50% in broiler starter diet. Ninety （90） day old chicks, of Anark breed were divided into three groups of three replicates each. There were ten birds per replicate and each group was fed each of the dietary treatment ad libitum for 28 days. Feed intake, body weight gain, feed conversion ratio （FCR）, protein efficiency ratio （PER）, total protein, albumin, globulin, aspartate aminotransaminase （EC 2.6.1.1.1）, alanine aminotransaminase （EC 2.6.1.1.2）, carcass cut parts and cost per kilogram flesh gained were monitored. Results showed linear increase （P 〈 0.05） in feed intake, body weight gain, FCR, and PER. The serum proteins and enzymes monitored were not influenced （P 〉 0.05） by treatments. The eviscerated weight, wings and large intestine length were higher （P 〈 0.05） in birds fed 50% replacement of maize diet. The cost of feed per kilogramme and cost per kilogramme flesh gained was best （P 〈 0.05） in birds fed diets 50% replacement of maize. Therefore, fermented mixture of cassava peel and caged layers＇ manure （FCPCLM） in broiler starter diet at 50% dietary level support good performance, enhanced gain in eviscerated weight and reasonable cost per kilogramme flesh gained at the starter phase.

Protective effect of Genistein against N-nitrosodiethylamine(NDEA)-induced hepatotoxicity in Swiss albino rats

In the present study,we studied the effect of Genistein against the hepatotoxicity induced by N-nitrosodiethylamine（NDEA）.NDEA is present in almost all kinds of food stuff and has been reported to be a hepatocarcinogen.The male rats were exposed to NDEA（0.1 mg/mL） dissolved in drinking water separately and along with 25,50,100 mg/mL of Genistein for 21 days.The activities of serum glutamic oxaloacetic transaminase（SGOT）,serum glutamic pyruvic transaminase（SGPT）.alkaline phosphatase（ALP） and lactate dehydrogenase（LDH） were measured in blood serum.Lipid peroxidation,protein carbonyl content,micronucleus frequency and DNA damage（Comet assay） were performed on rat hepatocytes.The results of the study reveal that the treatment of NDEA along with Genistein showed a significant dose-dependent decrease in the levels of blood serum enzymes i.e.,SGOT,SGPT,ALP and LDH（P〈0.05）.The HE staining of histological sections of the liver also revealed a protective effect of Genistein.A significant dose-dependent reduction in the lipid peroxidation and protein carbonyl content was observed in rats exposed to NDEA（0.1 mg/mL） along with Genistein（P〈0.05）.The results obtained for the comet assay in rat hepatocytes showed a significant dose-dependent decrease in the mean tail length（P〈0.05）.Thus the present study supports the hepatoprotective role of Genistein.

Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline

At 0.12 mmol/L γ-glutamyl p-nitroaniline(GGPNA),an improved integrated method was developed for kinetic analysis of γ-glutamyltransferase(GGT) reaction process and the integration with the classical initial rate method to measure serum GGT.For the improved integrated method,an integrated rate equation,which used the predictor variable of reaction time and considered inhibitions by both GGPNA and products,was nonlinearly fit to GGT reaction processes.For the integration strategy,classical initial rates were estimated when GGPNA consumption percentages were below 50%;otherwise,maximal reaction rates of GGT were estimated by the improved integrated method and converted into initial rates according to the differential rate equation at 0.11 mmol/L GGPNA.The inte-gration strategy was validated using optimized GGT kinetic parameters and 10-s intervals to record reaction curves within 8.0 min.By the integration strategy,there was a linear response from 0.9 to 32.0 U/L GGT,coefficients of variation were below 3.5%for GGT from 8.0 to 32.0 U/L(n=5) ,and GGT activities in clinical sera responded linearly to their classical initial rates at 2.00 mmol/L GGPNA with an expected slope.Therefore,the integration strategy was successful in measuring GGT at 0.12 mmol/L GGPNA.