Evaluation of polyherbal formulation in broilers fed high energy diet: Implications on zootechnical parameters, fat accretion, and serum L-carnitine levels.

Objective: The current broiler trial was planned to assess the effects of Kolin Plus™, a polyherbal formulation (PHF), on performance, protein and fat accretion, and serum L-carnitine (LC) levels in broilers fed a high-energy diet (HED). Materials and Methods: A total of 500 1-day-old Cobb 430 male chicks were assigned to 5 treatment groups consisting of 10 replicates, with 10 birds in each replicate (n = 100). Group G1 was a negative control fed HED, and group G2, a positive control supplemented with synthetic choline chloride (SCC) 1,500 gm/ton in HED. Groups G3, G4, and G5 were supplemented with PHF in HED at 400, 500, and 750 gm/ton feed, respectively (PHF400, PHF500, and PHF750). Results: The PHF produced a dose-dependent numerical improvement in body weight, feed conversion ratio, livability, and the European Production Index. There were no changes in carcass nitrogen and protein accretion, whereas a statistically significant decrease (p < 0.05) in carcass fat and fat accretion was observed in the SCC and PHF groups. Moreover, PHF showed a significant increase in serum LC levels. Conclusion: Kolin Plus™ improves performance parameters akin to SCC by improving fat metabolism and mobilization by enhancing serum LC levels and restoring normal fat accretion.

Effect of dietary supplementation of phytogenic feed additive on performance traits, serum neopterin, and cutaneous basophil hypersensitivity response in heat-induced stress model of broiler chickens.

OBJECTIVE: The trial was aimed at assessing the effect of phytogenic feed additive (PFA), a natural adaptogen, on growth performance, serum neopterin level, and cutaneous basophil hypersensitivity (CBH) response in heat-induced stress model of broilers. MATERIALS AND METHODS: One-day-old Ross 308 chicks (N = 360) were randomly distributed among normal control (NOR), heat-stress control (HSC), and PFA treatment (HSC plus PFA at 200 gm/ton of feed) group. HSC and PFA groups were subjected to heat stress (HS) (32°C-36°C) from 9:00 a.m. to 5:00 p.m. for 35 days. The impact of HS on growth performance, serum neopterin level, and CBH response was assessed. RESULTS: High ambient temperature worsened the performance traits [bodyweight (p < 0.05) and feed conversion ratio] and significantly lowered the serum neopterin level and CBH response in the HSC group when compared to the NOR group. However, supplementation of PFA at 200 gm/ton of feed to birds mitigated the detrimental effects of HS. CONCLUSION: PFA at 200 gm/ton demonstrated the immunomodulatory effect through the restoration of serum neopterin level, CBH response, and growth performance traits in heat-stressed broiler chickens. Thus, PFA can be used as a natural adaptogen to increase the stress resistance and mitigate the negative consequences of various stressors in broiler chickens.

Evaluation of a polyherbal formulation for the management of wet litter in broiler chickens: Implications on performance parameters, cecal moisture level, and footpad lesions.

OBJECTIVE: The study was carried out to develop a wet litter model with magnesium chloride to assess the effectiveness of a polyherbal formulation (PHF) on growth performance, litter and cecal moisture (LCM) level, cecal consistency (CC) score, and footpad lesions (FPLs) score in Ross 308 broiler chickens. MATERIALS AND METHODS: 1,200 one-day-old chicks were assigned into five groups: normal control, negative control [NTC; treated with 1.7% magnesium chloride hexahydrate (MgCl2.6H2O)], and three treatment groups, T1, T2, and T3, where 750, 1,000, and 2,000 gm/ton of PHF, respectively, were supplemented. All the groups were fed a basal diet until day 7. However, the NTC and treatment groups were fed a diet with MgCl2 from days 8 to 42. RESULTS: The addition of MgCl2 for 35 days worsened the growth performance traits in broilers and induced wet litter problems (FPL, high LCM, and poor CC) in the NTC group. However, PHF (750, 1,000, and 2,000 gm/ton) ameliorated the negative effect of a diet with MgCl2 on growth performance and wet litter problems, but a better response with respect to LCM and CC was observed in 2,000 gm/ton of PHF group, followed by that in 1,000 gm/ton of PHF group and 750 gm/ton of PHF group on day 42. CONCLUSION: The wet litter broiler model was developed through excessive feeding of MgCl2, which caused the performance parameters to worsen and the emergence of problems associated with the wet litter. Supplementation with PHF ameliorated these problems and, therefore, it can be used for the management of wet litter in poultry.

Mutagenicity and Acute Oral Toxicity Test for Herbal Poultry Feed Supplements.

Herbal products are being used and trusted globally for thousands of years for their health benefits and limited side effects. Globally, a general belief amongst the consumers is that herbal supplements are always safe because they are "natural." But later, research reveals that they may not be safe. This raises concern on their safety and implications for their use as feed supplement or medicine. Toxicity testing can reveal some of the risks that may be associated with use of herbs, therefore avoiding potential harmful effects. The present study was designed to investigate five poultry feed supplements (PFS), EGMAX® (to revitalize ovarian activity), FEED-X™ (feed efficiency enhancer), KOLIN PLUS™ (natural replacer of synthetic choline chloride), PHYTOCEE® (natural defence enhancer), and STODI® (to prevent and control loose droppings), for their possible mutagenicity and toxicity. Bacterial reverse mutation (BRMT) and acute oral toxicity tests were employed to assess the PFS for their possible mutagenicity and toxicity. Results indicated that the PFS were devoid of mutagenic effects in BRMT and showed higher safety profile in rodent acute oral toxicity test.

Evaluation of polyherbal formulation and synthetic choline chloride on choline deficiency model in broilers: implications on zootechnical parameters, serum biochemistry and liver histopathology.

OBJECTIVE: The study was designed to establish choline deficiency model (CDM) in broilers for evaluating efficacy of polyherbal formulation (PHF) in comparison with synthetic choline chloride (SCC). METHODS: A total of 2,550 one-day-old Cobb 430 broiler chicks were randomly assigned to different groups in three experiments. In experiment 1, G1 and G2 served as normal controls and were fed a basal diet with 100% soybean meal (SBM) as a major protein source supplemented with and without SCC, respectively. In G3, G4, G5, and G6 groups, SBM was replaced at 25%, 50%, 75%, and 100% by soy protein isolate (SPI) to induce a graded level of choline deficiency. In experiment 2, PHF (500 and 1,000 g/ton) in comparison with SCC (1,000 g/ton) were evaluated. In experiment 3, dose-response of PHF (200, 400, and 500 g/ton) with SCC (400 g/ton) was determined. RESULTS: Replacement of SBM by SPI produced a linear decrease in body weight gain (BWG) with a poor feed conversion ratio (FCR). 25% SBM replacement by SPI yielded an optimum negative impact on BWG and FCR; hence, it is considered for further studies. In experiment 2, PHF (500 and 1,000 g/ton) and SCC (1,000 g/ton) showed a similar performance in BWG, FCR and relative liver weight. In experiment 3, PHF produced an optimum efficacy at 400 g/ton and was comparable to SCC in the restoration of serum aspartate aminotransferase activity, abdominal fat, breast muscle lipid content and liver histopathological abnormalities. CONCLUSION: Replacement of SBM by SPI caused choline deficiency characterised by worsening of BWG, FCR, elevation in liver enzymes and histopathological changes indicating fatty liver. CDM was found valid for evaluating SCC and PHF. It is concluded that PHF has the potential to mimic biological activities of SCC through the restoration of negative effects caused by CDM.

Alleviation of Heat Stress by a Polyherbal Formulation, Phytocee™: Impact on Zootechnical Parameters, Cloacal Temperature, and Stress Markers.

BACKGROUND: The range of thermoneutral zone of chickens is narrow, and they become easily susceptible to environmental stress, a common and major concern for poultry causing a production loss. OBJECTIVE: The present study was designed to comparatively evaluate anti-stress activity of Phytocee™ and Vitamin C in chickens reared under heat stress. MATERIALS AND METHODS: A total of 600-day-old chicks of Cobb 400 were randomly assigned to 4 groups with 6 replicates comprising 25 birds each (n = 150). G1 served as a normal control (NC) and supplemented with Vitamin C at 100 g/ton of feed. G2 served as a heat stress control (HSC), subjected to heat stress (34°C-36°C) without Vitamin C supplementation. G3 and G4 served as positive control and treatment group (TC), subjected to heat stress and supplemented with Vitamin C and Phytocee™ at 100 g/ton of feed, respectively. The impact on zootechnical parameters and cloacal temperature was assessed at regular intervals, and blood was collected at the end of the experiment for evaluation of stress parameters, namely heterophil lymphocyte ratio (H:L ratio) and serum corticosterone. RESULTS: Exposure of chickens to heat stress caused a significant decrease in body weight, worsening of feed conversion ratio, higher mortality, and poor production efficiency. Moreover, serum corticosterone level, H:L ratio, and cloacal temperature were significantly increased in HSC as compared to NC. However, supplementation of Phytocee™ in feed significantly ameliorated the negative impact of heat stress in broiler birds. CONCLUSION: The supplementation of Phytocee™ demonstrated an anti-stress effect in chickens through restoration of serum corticosterone level, H:L ratio, and thermoregulatory mechanism. SUMMARY: Combating heat stress remains a challenge for the broiler industry in the tropics and subtropics, which is even aggravated by the changing climatic conditionsThe present study was designed to evaluate the anti-stressor activity of Phytocee™, a polyherbal formulation containing Emblica officinalis, Ocimum sanctum, and Withania somnifera in broiler using heat stress model in comparison with Vitamin CPhytocee™ demonstrated an anti-stress effect in the current study by ameliorating the negative effects of heat stress on zootechnical parameters, serum corticosterone, heterophil lymphocyte ratio, and cloacal temperature of broilers through modulating the hypothalamic-pituitary-adrenal axis and thermoregulatory mechanismHence, Phytocee™ could be recommended in broilers and livestock animals for modulating and combating adverse effects of heat stress and thereby reducing the economic losses incurred by farmers. Abbreviations Used: HPA axis: Hypothalamic pituitary adrenal axis.

Gender differences and protective effects of testosterone in collagen induced arthritis in rats.

To understand the gender differences noticed in autoimmune disorders, particularly rheumatoid arthritis, we used a rat model of collagen induced arthritis (CIA). This study was carried out in two parts. In the first study, severity of inflammation was compared between male and female rats with respect to radiology, histology, activities of lysosomal enzymes, lipid peroxidation, immune response to type II collagen and the level of prostaglandin, a major inflammatory mediator. Since female rats developed severe inflammation, this study was extended to confirm if testosterone at physiological concentration had protective effect against CIA. Hence, studies were carried out on the effect of testosterone application on castrated arthritic rats. Female arthritic rats were also treated with testosterone to find out the effectiveness of the androgen in the presence of female hormones. Results of this study conclusively showed that testosterone possessed significant anti-inflammatory effects at physiological concentration and exerted its action in a gender nonspecific manner.

Low frequency and low intensity pulsed electromagnetic field exerts its antiinflammatory effect through restoration of plasma membrane calcium ATPase activity.

Rheumatoid arthritis (RA) is a chronic inflammatory disorder affecting 1% of the population worldwide. Pulsed electromagnetic field (PEMF) has a number of well-documented physiological effects on cells and tissues including antiinflammatory effect. This study aims to explore the antiinflammatory effect of PEMF and its possible mechanism of action in amelioration of adjuvant induced arthritis (AIA). Arthritis was induced by a single intradermal injection of heat killed Mycobacterium tuberculosis at a concentration of 500 microg in 0.1 ml of paraffin oil into the right hind paw of rats. The arthritic animals showed a biphasic response regarding changes in the paw edema volume. During the chronic phase of the disease, arthritic animals showed an elevated level of lipid peroxides and depletion of antioxidant enzymes with significant radiological and histological changes. Besides, plasma membrane Ca(2+) ATPase (PMCA) activity was inhibited while intracellular Ca(2+) level as well as prostaglandin E(2) levels was noticed to be elevated in blood lymphocytes of arthritic rats. Exposure of arthritic rats to PEMF at 5 Hzx4 microT x 90 min, produced significant antiexudative effect resulting in the restoration of the altered parameters. The antiinflammatory effect could be partially mediated through the stabilizing action of PEMF on membranes as reflected by the restoration of PMCA and intracellular Ca(2+) levels in blood lymphocytes subsequently inhibiting PGE(2) biosynthesis. The results of this study indicated that PEMF could be developed as a potential therapy for RA in human beings.

Alterations in band 3 protein and anion exchange in red blood cells of renal failure patients.

The precise nature of band 3 protein and its involvement in oxalate exchange in the red blood cells (RBCs) of renal failure patients has not been studied in detail. Therefore, here we studied the oxalate exchange and binding by band 3 protein in RBCs of humans with conditions of acute and chronic renal failure (ARF and CRF). The RBCs of ARF and CRF patients exhibited abnormal red cell morphology and an increased resistance to osmotic hemolysis. Further, an increase in the cholesterol content and decrease in the activities of Na(+)-K(+)-, Ca(2+)-, and Mg(2+)-ATPases of membranes were observed in the RBCs of ARF and CRF patients. A decrease in the oxalate flux was observed in the RBCs of ARF and CRF patients. The oxalate-binding activities of the RBC membranes were significantly lower in ARF (20 pmoles/mg protein) and CRF (5.3 pmoles/mg protein) patients as compared to that in the normal subjects (36 pmoles/mg protein). DEAE-cellulose and Sephadex G-200 column chromatography purification profiles revealed a distinctive shift in oxalate-binding activity of band 3 protein of RBCs of ARF and CRF patients as compared to that of the normal subjects. It was also observed from the binding studies with a fluorescent dye, eosin-5-maleimide, which specifically binds to band 3 protein, that the RBCs of ARF and CRF patients exhibited only 53 and 32% of abundance of band 3 protein, respectively, as compared to that in the RBCs of the normal subjects, thus revealing a decrease in the band 3 protein content in ARF and CRF patients. These results for the first time showed a decrease in the oxalate exchange in RBCs of patients with ARF and CRF, which was also concomitant with the low levels of abundance of band 3 protein.

Supplementation of vitamin E and selenium prevents hyperoxaluria in experimental urolithic rats.

Renal injury is considered as one of the prerequisites for calcium oxalate retention. In order to determine the role of lipid peroxidation related effects for hyperoxaluria, we evaluated the alterations in lipid peroxidation, antioxidants and oxalate synthesizing enzymes in lithogenic rats with response to vitamin E + selenium treatment. In kidney of lithogenic rats, the level of lipid peroxidation and the activities of oxalate synthesizing enzymes were found to be increased whereas the levels/activities of non-enzymatic and enzymatic antioxidants were found to be decreased. The urinary excretion of both oxalate and calcium were significantly elevated. Supplementation of lithogenic rats with vitamin E + selenium decreased the levels of lipid peroxides and the activities of oxalate synthesizing enzymes like glycolic acid oxidase (GAO), lactate dehydrogenase (LDH), xanthine oxidase (XO) with a concomitant increase in the activities of enzymatic antioxidants like superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glucose-6-phosphate dehydrogenase (G6PDH) and increased levels of non-enzymatic antioxidants like ascorbic acid, alpha-tocopherol and reduced glutathione (GSH). The urinary excretion of oxalate and calcium were normalized. The antioxidants vitamin E + selenium thereby protected from hyperoxaluria.

Oxalate binding proteins in calcium oxalate nephrolithiasis.

The existence of several oxalate specific binding proteins have been demonstrated in human and rat kidney. These occur in both cortical and medullary cells and are distributed mostly in the subcellular organelles. About 1/3 of the total cellular oxalate binding was localised in the inner mitochondrial membrane while the rest was in the nucleus. The purified mitochondrial oxalate binding protein (62 kDa) was composed, with a higher molar proportion, of basic amino acids, and could accumulate oxalate on incorporation into liposomes. In the nucleus, histone H(1B) (27.5 kDa), nuclear membrane protein (68 kDa) and nuclear pore complex protein (205 kDa) were present with oxalate binding activities. In addition, a 45 kDa calcium oxalate binding protein was identified in most of the subcellular organelles. Both mitochondrial and nuclear oxalate binding proteins and calcium oxalate binding protein have shown the kinetic properties of specificity, saturability, pH and temperature dependency, energy of activation and inhibition by substrate analogues. All oxalate binding proteins were sensitive to the transport inhibitor 4'-4' diisothiocyano stilbene-2-2 disulphonic acid (DIDS), which is known to interact with the lysine moiety of the proteins. Calcium oxalate monohydrate (COM) crystals adsorbed oxalate binding proteins from human and rat kidney, and oxalate binding proteins isolated from human kidney stone matrix also exhibited the above kinetic properties. In experimental hyperoxaluria, all of the renal oxalate binding proteins showed enhanced oxalate binding activity with increased protein concentration. This enhanced oxalate binding activity was also attributed to increased lipid peroxidation, which correlated positively, and to decreased thiol status, which correlated negatively. A positive correlation was observed between the lipid peroxidation and both the oxalate binding activity of the in vitro peroxidised subcellular organelles and the purified protein. Similarly, in an in vivo hyperoxaluric condition, a negative correlation was observed between thiol content and both the oxalate binding activity of the peroxidised subcellular organelles and the purified protein. In the calcium oxalate crystal growth system, the oxalate binding proteins behaved either as promoters or inhibitors of the nucleation and aggregation of crystals. Following the peroxidation of the proteins, the degree of effect of the promoter protein was further stimulated while the degree of inhibition caused by the inhibitor protein further declined. Similar observations were duplicated with the proteins derived from hyperoxaluric rat kidney or kidney homogenate subjected to in vitro lipid peroxidation. The oxalate binding proteins were thought to modulate the crystallisation process in an hyperoxaluric condition similar to calcium specific binding protein modulators.

Effect of cyclosporin A treatment on renal calcium oxalate binding in experimental hyperoxaluria.

This study was aimed to investigate the effect ofCyclosporin A administration on renal calcium oxalate binding under hyperoxaluric condition. Cyclosporin A administration or ammonium oxalate treatment increased calcium oxalate binding, which was further increased in kidney treated with cyclosporin A and ammonium oxalate together. The increase of calcium oxalate binding was associated with lipid peroxidation as well as with a concomitant decrease in total thiol in both rat and human kdiney homogenate. Cyclosporin A administration to hyperoxaluric rats resulted with increased calcium oxalate binding protein. However there was no change with specific activity of the protein. In conclusion, Cyclosporin A administration either to normal or hyperoxaluric rats is resulted with increased concentration of calcium oxalate binding protein as well as enhanced activity due to membrane lipid peroxidation.

Calcium oxalate stone disease: role of lipid peroxidation and antioxidants.

Membrane injury facilitated the fixation of calcium oxalate crystals and subsequent growth into kidney stones. Oxalate-induced membrane injury was mediated by lipid peroxidation reaction through the generation of oxygen free radicals. In urolithic rat kidney or oxalate exposed cultured cells, both superoxide anion and hydroxyl radicals were generated in excess, causing cellular injury. In hyperoxaluric rat kidney, both superoxide and H2O2-generating enzymes such as glycolic acid oxidase (GAO) and xanthine oxidase (XO) were increased, and hydroxyl radical and transition metal ions, iron, and copper were accumulated. The lipid peroxidation products, thiobarbituric acid-reactive substances (TBARS), hydroperoxides, and diene conjugates were excessively released in tissues of urolithic rats and in plasma of rats as well as stone patients. The accumulation of these products was concomitant with the decrease in the antioxidant enzymes, superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and glucose-6 phosphate dehydrogenase (G6PD) as well as radical scavengers, vitamin E, ascorbic acid, reduced glutathione (GSH), and protein thiol. All the above parameters were decreased in urolithic condition, irrespective of the agents used for the induction of urolithiasis. Oxalate binding activity and calcium oxalate crystal deposition were markedly pronounced, along with decreased adenosine triphosphatase (ATPase) activity. Lipid peroxidation positively correlated with cellular oxalate, oxalate binding, gamma-glutamyl carboxylase, and calcium level and negatively correlated with GSH, vitamin E. ascorbic acid, and total protein thiol. Antioxidant therapy to urolithic rats with vitamin E, glutathione monoester, methionine, lipoic acid, or fish oil normalised the cellular antioxidant system, enzymes and scavengers, and interrupted membrane lipid and protein peroxidation reaction, ATPase inactivation, and its associated calcium accumulation. Antioxidant therapy prevented calcium oxalate precipitation in the rat kidney and reduced oxalate excretion in stone patients. Similarly, calcium oxalate crystal deposition in vitro to urothelium was prevented by free radical scavengers such as phytic acid and mannitol by protecting the membrane from free radical-mediated damage. All these observations were suggestive of the active involvement of free radical-mediated lipid peroxidation-induced membrane damage in the pathogenesis of calcium oxalate crystal deposition and retention.

Effect of hyperoxaluria on the inhibitory activity of a 45-kD urinary protein.

Proteins are thought to play a major role in stone formation and structurally abnormal proteins have been reported to be present in the urine of stone formers. This study was aimed to determine whether hyperoxaluria modifies the kinetic properties of urinary inhibitory proteins. Hyperoxaluria was induced by feeding 1% ethylene glycol to rats. Oxalate, uric acid and calcium excretion were increased progressively during hyperoxaluria, while magnesium level was decreased. Urinary proteins were separated on a DEAE-cellulose column by eluting with stepwise increasing salt concentration in 0.05 M Tris-HCl buffer (pH 7.0). Each protein fraction was studied for its crystallization inhibitory potential by the spectrophotometric method. The protein eluted in 0.3 M NaCl containing buffer had the maximal nucleation as well as inhibitory activity. The protein had a molecular weight of 45 kD. In hyperoxaluria, the urinary excretion of this protein significantly increased. In the crystal growth assay, the control rat 45-kD protein inhibited nucleation by 75% and aggregation by 100%. In contrast, it is very interesting to note that the protein derived from 28th day hyperoxaluric urine, behaved as a promoter of nucleation (-113%, percentage inhibition) and weak inhibitor of aggregation (28%). A significantly high negative correlation (r = -0.97) between oxalate excretion and the inhibitory activity of the 45-kD protein was observed suggesting a modification of the protein by oxalate.