Do little embryos make big decisions? How maternal dietary protein restriction can permanently change an embryo's potential, affecting adult health

Periconceptional environment may influence embryo development, ultimately affecting adult health. Here, we review the rodent model of maternal low-protein diet specifically during the preimplantation period (Emb-LPD) with normal nutrition during subsequent gestation and postnatally. This model, studied mainly in the mouse, leads to cardiovascular, metabolic and behavioural disease in adult offspring, with females more susceptible. We evaluate the sequence of events from diet administration that may lead to adult disease. Emb-LPD changes maternal serum and/or uterine fluid metabolite composition, notably with reduced insulin and branched-chain amino acids. This is sensed by blastocysts through reduced mammalian target of rapamycin complex 1 signalling. Embryos respond by permanently changing the pattern of development of their extra-embryonic lineages, trophectoderm and primitive endoderm, to enhance maternal nutrient retrieval during subsequent gestation. These compensatory changes include stimulation in proliferation, endocytosis and cellular motility, and epigenetic mechanisms underlying them are being identified. Collectively, these responses act to protect fetal growth and likely contribute to offspring competitive fitness. However, the resulting growth adversely affects long-term health because perinatal weight positively correlates with adult disease risk. We argue that periconception environmental responses reflect developmental plasticity and 'decisions' made by embryos to optimise their own development, but with lasting consequences.

Identification of DNA-binding proteins by incorporating evolutionary information into pseudo amino acid composition via the top-n-gram approach

DNA-binding proteins are crucial for various cellular processes and hence have become an important target for both basic research and drug development. With the avalanche of protein sequences generated in the postgenomic age, it is highly desired to establish an automated method for rapidly and accurately identifying DNA-binding proteins based on their sequence information alone. Owing to the fact that all biological species have developed beginning from a very limited number of ancestral species, it is important to take into account the evolutionary information in developing such a high-throughput tool. In view of this, a new predictor was proposed by incorporating the evolutionary information into the general form of pseudo amino acid composition via the top-n-gram approach. It was observed by comparing the new predictor with the existing methods via both jackknife test and independent data-set test that the new predictor outperformed its counterparts. It is anticipated that the new predictor may become a useful vehicle for identifying DNA-binding proteins. It has not escaped our notice that the novel approach to extract evolutionary information into the formulation of statistical samples can be used to identify many other protein attributes as well.

Stream dissolved organic matter bioavailability and composition in watersheds underlain with discontinuous permafrost

We examined the impact of permafrost on dissolved organic matter (DOM) composition in Caribou-Poker Creeks Research Watershed (CPCRW), a watershed underlain with discontinuous permafrost, in interior Alaska. We analyzed long term data from watersheds underlain with varying degrees of permafrost, sampled springs and thermokarsts, used fluorescence spectroscopy, and measured the bioavailabity of dissolved organic carbon (DOC). Permafrost driven patterns in hydrology and vegetation influenced DOM patterns in streams, with the stream draining the high permafrost watershed having higher DOC and dissolved organic nitrogen (DON) concentrations, higher DOC:- DON and greater specific ultraviolet absorbance (SUVA) than the streams draining the low and medium permafrost watersheds. Streams, springs and thermokarsts exhibited a wide range of DOC and DON concentrations (1.5–37.5 mgC/L and 0.14–1.26 mgN/L, respectively), DOC:DON (7.1–42.8) and SUVA (1.5–4.7 L mgC-1 m-1). All sites had a high proportion of humic components, a low proportion of protein components, and a low fluorescence index value (1.3–1.4), generally consistent with terrestrially derivedDOM. Principal component analysis revealed distinct groups in our fluorescence data determined by diagenetic processing and DOM source. The proportion of bioavailable DOC ranged from 2 to 35%, with the proportion of tyrosine- and tryptophan-like fluorophores in the DOM being a major predictor of DOC loss (p\0.05, R2 = 0.99). Our results indicate that the degradation of permafrost in CPCRW will result in a decrease in DOC and DON concentrations, a decline in DOC:DON, and a reduction in SUVA, possibly accompanied by

Suppression of nuclear factor-κB activity in macrophages by chylomicron remnants: modulation by the fatty acid composition of the particles

Current evidence indicates that chylomicron remnants (CMR) induce macrophage foam cell formation, an early event in atherosclerosis. Inflammation also plays a part in atherogenesis and the transcription factor nuclear factor-kappaB (NF-kappaB) has been implicated. In this study, the influence of CMR on the activity of NF-kappaB in macrophages and its modulation by the fatty acid composition of the particles were investigated using macrophages derived from the human monocyte cell line THP-1 and CMR-like particles (CRLPs). Incubation of THP-1 macrophages with CRLPs caused decreased NF-kappaB activation and downregulated the expression of phospho-p65-NF-kappaB and phospho-IkappaBalpha (pIkappaBalpha). Secretion of the inflammatory cytokines tumour necrosis factor alpha, interleukin-6 and monocyte chemoattractant protein-1, which are under NF-kappaB transcriptional control, was inhibited and mRNA expression for cyclooxygenase-2, an NF-kappaB target gene, was reduced. CRLPs enriched in polyunsaturated fatty acids compared with saturated or monounsaturated fatty acids had a markedly greater inhibitory effect on NF-kappaB binding to DNA and the expression of phospho-p65-NF-kappaB and pIkappaB. Lipid loading of macrophages with CRLPs enriched in polyunsaturated fatty acids compared with monounsaturated fatty acids or saturated fatty acids also increased the subsequent rate of cholesterol efflux, an effect which may be linked to the inhibition of NF-kappaB activity. These findings demonstrate that CMR suppress NF-kappaB activity in macrophages, and that this effect is modulated by their fatty acid composition. This downregulation of inflammatory processes in macrophages may represent a protective effect of CMR which is enhanced by dietary polyunsaturated fatty acids.

The role of glucocorticoids in naturally fasting grey seal (Halichoerus grypus) pups: dexamethasone stimulates mass loss and protein utilisation, but not departure from the colony

Seals must manage their energy reserves carefully while they fast on land to ensure that they go to sea with sufficient fuel to sustain them until they find food. Glucocorticoids (GCs) have been implicated in the control of fuel metabolism and termination of fasting in pinnipeds. Here we tested the hypothesis that dexamethasone, an artificial GC, increases fat and protein catabolism, and induces departure from the breeding colony in wild, fasting grey seal pups. A single intramuscular dose of dexamethasone completely suppressed cortisol production for 24–72 h, demonstrating activation of GC receptors. In experiment 1, we compared the effects of a single dose of dexamethasone or saline administered 10 days after weaning on fasting mass and body composition changes, cortisol, blood urea nitrogen (BUN) and glucose levels, and timing of departure from the colony. In experiment 2, we investigated the effects of dexamethasone on short-term (5 days) changes in mass loss, body composition and BUN levels. In experiment 1, dexamethasone induced a short-lived increase in mass loss, but there was no difference in timing of departure between dexamethasone- and saline-treated pups (N=10). In experiment 2, dexamethasone increased protein and water loss and prevented a decrease in BUN levels (N=11). Our data suggest changes in cortisol contribute to regulation of protein catabolism in fasting seal pups, irrespective of the sex of the animal, but do not terminate fasting. By affecting the rate of protein depletion, lasting changes in cortisol levels could influence the amount of time seal pups have to find food, and thus may have important consequences for their survival.

Effects of mass and body composition on fasting fuel utilisation in grey seal pups (Halichoerus grypus Fabricius): an experimental study using supplementary feeding

This study used supplementary feeding to test the hypothesis that fuel partitioning during the postweaning fast in grey seal pups is affected by size and composition of energy reserves at weaning, and by extra provisioning. Mass and body composition changes were measured during suckling and fasting to investigate the effect of natural differences in energy reserves at weaning on subsequent allocation of fat and protein to energy use. We fed seven pups for 5 days after weaning, to investigate the effect of increased fuel availability, and particularly protein, on fuel utilisation. After correcting for protein used during the moult, the proportional contribution of fat was 86–99% of total energy use. Pups with greater energy reserves, i.e. those that were heavier and fatter at weaning, had higher rates of fat and energy use. There was no significant relationship between adiposity at weaning and proportional contribution of fat to energy use, perhaps due to a limited sample size or range of body masses and adiposity. Supplemented individuals used energy, specifically fat, much faster and utilised proportionally less of their endogenous protein by departure than non-supplemented individuals. Fat metabolism contributed a similar percentage to daily energy use in both groups. These findings show that pups spare protein, even when energy use is dramatically increased. Pups that receive greater maternal provisioning and lay down more protein may have increased survival chances at sea. This study highlights the importance of protein reserves in first year survival of grey seal pups.

Relations entre la composition du lait et les facteurs alimentaires dans les troupeaux laitiers québécois

L’objectif général des travaux présentes dans cette thèse de doctorat était d’établir la relation entre l’alimentation et la composition du lait des vaches laitières, en mettant l’emphase sur les teneurs en matières grasses (TMG), sur l’urée du lait et sur l’efficience d’utilisation de l’azote. Pour la TMG, c’est principalement la relation entre cette teneur et la différence alimentaire cations-anions (DACA) qui a été investiguée. Une base de données de 2 142 troupeaux québécois a été utilisée et la relation entre la composition de la ration, incluant la DACA, et la TMG du lait a été déterminée à l’aide de régressions multiples. Il a été possible de prédire entre 32 et 66 % de la variation de la TMG du lait en fonction du stade de lactation. Malgré plusieurs interactions trouvées, une augmentation de la DACA, une supplémentation avec de l’acide palmitique et une distribution des aliments en ration totale mélangée ont eu une relation positive avec la TMG du lait, tandis qu’une augmentation de la proportion de concentrés dans la ration a eu un effet négatif. Les modèles développés ont montré l’importance de la gestion de l’alimentation sur la TMG du lait. En plus, ils ont démontré l’intérêt de considérer la DACA dans la formulation de rations chez la vache laitière. Dans une deuxième étude, la même base des données a été utilisée pour identifier les facteurs nutritionnels qui peuvent faire varier la teneur en urée du lait. Contrairement à ce qui est mentionné dans la littérature, tant des corrélations positives et que négatives entre les teneurs en urée du lait et en protéines des rations à l’intérieur des troupeaux sur une période de 36 mois ont été obtenues. Pour mieux comprendre ces relations, les résultats de performances et d’alimentation de 100 troupeaux avec des corrélations positives (r > 0,69) et de 100 troupeaux avec des corrélations négatives (r < -0,44) entre la teneur en urée du lait et en protéine brute de la ration ont été comparés. Les résultats n’ont pas montré de différences entre les deux groupes ni pour la composition de la ration, ni pour l’urée du lait. Ces résultats ne permettent pas d’identifier le meilleur contexte pour l’utilisation de la teneur en urée du lait comme un outil de gestion de l’alimentation. Ces observations soulèvent des questions sur la validité de l’utilisation des statistiques d’alimentation provenant de la base de données utilisée pour des évaluations nutritionnelles plus spécifiques. Considérant les résultats du projet précédent, le dernier projet visait à mieux comprendre les caractéristiques des fermes avec différentes efficiences d’utilisation de l’azote en utilisant des données plus fiables. Ainsi, 100 fermes laitières au Québec ont été visitées pour recueillir les données de production, de consommation d’aliments et de gestion de leur troupeau. Ces fermes ont été divisées en quatre groupes par une analyse en grappes selon leur efficience d’utilisation de l’azote. La comparaison entre les groupes a montré que les fermes dans les groupes avec une plus haute efficience d’utilisation d’azote ont une production laitière moyenne par vache plus élevée. Pour les stratégies d’alimentation, les fermes plus efficientes donnent plus d’énergie, mais moins de protéines brutes que les fermes des groupes moins efficients. Les résultats ont également montré l’importance de la prise alimentaire des vaches sur l’efficience d’utilisation de l’azote puisque les fermes des groupes avec la plus grande efficience étaient également celles avec la plus faible prise alimentaire. Aussi, les résultats n’ont pas permis d’établir clairement la relation entre la teneur en urée du lait et l’efficience de l’utilisation de l’azote. En effet, des valeurs différentes pour l’urée du lait étaient obtenues entre le groupe plus efficient et le moins efficient, mais la faible ampleur de variation de l’efficience d’utilisation de l’azote des groupes intermédiaires n’a pas permis d’observer de différences pour l’urée du lait. Finalement, outre une réduction des risques de pollution, les fermes des groupes plus efficaces pour l’utilisation de l’azote étaient également celles avec la marge sur les coûts d’alimentation par les vaches plus élevées. Par conséquent, il y a aussi un intérêt économique à améliorer l’efficience de l’utilisation de l’azote sur les fermes.

Proximate composition and mineral contents of pebbly fish, Alestes baremoze (Joannis, 1835) fillets in relation to fish size

Alestes baremoze (Joannis, 1835), locally known as Angara in Uganda, is native to fresh water systems in Africa, thriving well in both lacustrine and riverine conditions. It is part of the routine diets of families in northern Uganda, South Sudan, the Sudan and the Democratic Republic of Congo. The objective of this study was to determine the proximate composition and mineral contents of A. baremoze fillets according to fish size. The mineral contents of A. baremoze from Lake Albert were analysed using standard procedures. The fish samples were categorised into three size-groups; <1 kg (880–990g), 1-1.5 kg and 1.6-2.5 kg. On wet weight basis, there were no significant differences (p>0.05) in crude protein and ash content among the different fish sizes. However, there were significant differences (p<0.05) in crude fat, carbohydrate, gross energy and vitamin A. Crude fat (0.35%), carbohydrate (0.37%) and gross energy (597.6 Kcal/100 g) were significantly higher in medium sized fish (1 to 1.5 kg) compared with the larger fish category. Vitamin A contents of different fish sizes ranged from 55.1 to75.3 μg RAE/100g. The contents of magnesium and iron were highest in sizes <1 kg (5.34 mg/100 g) and (3.58 mg/100 g), respectively. It was observed that potassium content (339.33 mg/100 g) and calcium (29.75 mg/100 g) were significantly higher (p<0.05) in fish >1.5 kg. These findings suggest that taste, freshness and other related external appearances should not be the only factors to be considered in making choice for marketing and consumption of Alestes baremoze.

Supplementation of cattle fed tropical grasses with microalgae increases microbial protein production and average daily gain

A series of 3 experiments were conducted to evaluate the use of microalgae as supplements for ruminants consuming low-CP tropical grasses. In Exp. 1, the chemical composition and in vitro protein degradability of 9 algae species and 4 protein supplements were determined. In Exp. 2, rumen function and microbial protein (MCP) production were determined in Bos indicus steers fed speargrass hay alone or supplemented with Spirulina platensis, Chlorella pyrenoidosa, Dunaliella salina, or cottonseed meal (CSM). In Exp. 3, DMI and ADG were determined in B. indicus steers fed speargrass hay alone or supplemented with increasing amounts of NPN (urea combined with ammonia sulfate), CSM, or S. platensis. In Exp. 1, the CP content of S. platensis and C. pyrenoidosa (675 and 580 g/kg DM) was highest among the algae species and higher than the other protein supplements evaluated, and Schizochytrium sp. had the highest crude lipid (CL) content (198 g/kg DM). In Exp. 2, S. platensis supplementation increased speargrass hay intake, the efficiency of MCP production, the fractional outflow rate of digesta from the rumen, the concentration of NH3N, and the molar proportion of branched-chain fatty acids in the rumen fluid of steers above all other treatments. Dunaliella salina acceptance by steers was low and this resulted in no significant difference to unsupplemented steers for all parameters measured for this algae supplement. In Exp. 3, ADG linearly increased with increasing supplementary N intake from both S. platensis and NPN, with no difference between the 2 supplements. In contrast, ADG quadratically increased with increasing supplementary N intake from CSM. It was concluded that S. platensis and C. pyrenoidosa may potentially be used as protein sources for cattle grazing low-CP pastures.

Special low protein foods for phenylketonuria: availability in Europe and an examination of their nutritional profile

Background Special low protein foods (SLPF) are essential in the nutritional management of patients with phenylketonuria (PKU). The study objectives were to: 1) identify the number of SLPF available for use in eight European countries and Turkey and 2) analyse the nutritional composition of SLPF available in one of these countries. Methods European Nutritionist Expert Panel on PKU (ENEP) members (Portugal, Spain, Belgium, Italy, Germany, Netherlands, UK, Denmark and Turkey) provided data on SPLF available in each country. The nutritional composition of Portuguese SLPF was compared with regular food products. Results The number of different SLPF available in each country varied widely with a median of 107 [ranging from 73 (Portugal) and 256 (Italy)]. Food analysis of SLPF available from a single country (Portugal) indicated that the mean phenylalanine content was higher in low protein baby cereals (mean 48 mg/100 g) and chocolate/energy bars/jelly (mean 41 mg/100 g). The energy content of different foods from a sub-group of SLPF (cookies) varied widely between 23 and 96 kcal/cookie. Low protein bread had a high fat content [mean 5.8 g/100 g (range 3.7 to 10)] compared with 1.6 g/100 g in regular bread. Seven of the 12 SLPF sub-groups (58 %) did not declare any vitamin content, and only 4 (33 %) identified a limited number of minerals. Conclusions Whilst equal and free access to all SLPF is desirable, the widely variable nutritional composition requires careful nutritional knowledge of all products when prescribed for individual patients with PKU. There is a need for more specific nutritional standards for special low protein foods.

Ionic composition of endolymph in teleosts: Origin and importance of endolymph alkalinity

Ionic (Na+, K+, Cl-, PO43-, pH), total CO2, total calcium and protein concentrations in the plasma and endolymph of the inner ear were compared in trout Oncorhynchus mykiss and turbot Scophthalmus maximus. In both species, saccular endolymph was characterized by high levels of K+ and total CO2 and in trout by an alkaline pH, The kinetic characteristics of proton secretion across the saccular epithelium of trout were investigated using a titration technique in which isolated saccules were mounted as closed sacs. The rate of proton secretion depends strongly on the pH of the Ringer's solution and secretion stops at a pH below 7.2, Proton secretion is driven by an energy-dependent mechanism involving basolateral ouabain-sensitive Na+/K+ exchangers. Proton secretion was partially inhibited by acetazolamide and completely inhibited in Na+-free Ringer or in the presence of 1 mmol l(-1) amiloride. A cellular model stressing the importance of proton exchange through the saccular epithelium is proposed to explain the regulation of endolymph pH, a crucial factor for the deposition of otolith calcium.

Chemical composition of Boletus pinophilus and Clitocybe subconnexa: preservation with gamma irradiation

The short shelf life of mushrooms is a barrier for their distribution and, therefore, there has been extensive research to find technologies that ensure the preservation of mushrooms, maintaining their organoleptic and nutritional properties (1]. Irradiation has proved its technological feasibility to be safely used in the reduction of food losses, being recognized by international organizations as a valid conservation alternative in extending shelflife of many foods. The aim of the present work was to validate the use of 2 kGy dose of gamma radiation to maintain chemical composition of wild mushrooms. Boletus pinophilus Pihit & Dermek and Clitocybe subconnexa Murrill wild samples were obtained in Tnis-os-Montes; subsequently, the samples were divided in two groups: control (non-irradiated, 0 kGy) and irradiated (2 kGy). The irradiation of the samples was performed in a 6°Co experimental chamber. Moisture, protein, fat, carbohydrates and ash were determined following the standard procedures [2]. Free sugars and tocopherols were determined by high performance liquid chromatography coupled to a refraction index detector (HPLC-RI) and a fluorescence detector, respectively; fatty acids were determined by gas-liquid chromatography with flame ionization detection (GC-FID) [3]. The protein and ash content was preserved in both groups, although the sugars and tocopherols decreased in the irradiated samples. Sugars and fatty acids showed significant changes after irradiation treatment, particularly in B. pinophillus, nevertheless, the magnitude of the obtained differences did not seem to be sufficient to affect the chemical profiles of the assayed mushrooms. Overall, the detected chemical changes might be considered as allowable, in view of the high advantages offered by gamma irradiation at decontamination and/or disinfestation level.

NUTRITIONAL COMPOSITION AND ANTIOXIDANT PROPERTIES OF RAPHIONACME SPLENDENS (SCHL.) TUBERS

People of Western Kordofan (Sudan) are endowed with a deep knowledge concerning the use of wild plants. Tubers of Raphionacme splendens Schl. subspecies splendens Flickr (family Apocynaceae), locally known as Elfayo, are used as a food reserve during times of famine or poor harvest. The aim of this study was to analyze the nutritional composition and antioxidant capacity of root tubers of R. splendens. Samples were collected from South-West Kordofan. Analyses included determination of moisture, carbohydrate, crude protein, fat, fibre, ash, minerals, vitamin C, amino acids and fatty acids composition. Antioxidant activity was determined by the DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2.2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) assays. The total phenolic content was also assessed. The results, which are referred to as (%) dry weight, showed that the tubers contained 3.2% protein, 18% carbohydrate, 0.5% lipid, 2.4% crude fibre, 3.5% ash, 79.2% moisture and gross energy 101.7 kJ/g. The total amino acids were 10776 mg/100g where the essential amino acids represented 28.2%. The more abundant essential amino acids were leucine (792mg/100g), isoleucine (712mg/100g) and threonine (536mg/100g). Methionine and lysine were the limiting amino acids. Minerals were potassium (259mg/100g), calcium (183mg/100g), magnesium (64mg/100g), phosphorus (37mg/100g), copper (3.6mg/100g), manganese (2.4mg/100g), zinc (1.8mg/100g) and iron (1.2mg/100g). Total saturated fatty acids were 45.6% whereas total unsaturated fatty acids were 54.4%. Oleic acid (32.56%) and palmitic acid (30.23%) were the most abundant fatty acids. Tubers displayed good antioxidant activity with IC50 values 0.987 and 1.559mg/mL against DPPH and ABTS radicals respectively. Vitamin C was 31.5mg/100g and total phenolic content was 60mg gallic acid equivalent (GAE) per 100g dry sample and they could be the main contributor to the antioxidant capacity of the tubers. In conclusion, the results of this study suggested that tubers of R. splendens could have beneficial effect for food and/or nutraceutical application for normal growth and adequate protection against diseases associated with reactions of free radicals.

Chemical composition of cultivated seaweed ulva clathrata (Roth) C. Agardh

Samples of cultivated Ulva clathrata were collected from a medium scale system (MSS, 1.5 1.5 m tank), or from a large scale system (LSS, 0.8 ha earthen pond). MSS samples were dried directly while the LSS sample was washed in freshwater and pressed before drying. Crude protein content ranged 20–26%, essential amino acids accounting for 32–36% of crude protein. The main analysed monosaccharides were rhamnose (36–40%), uronic acids (27–29%), xylose (10–13%) and glucose (10–16%). Some notable variations between MSS and LSS samples were observed for total dietary fibre (26% vs 41%), saturated fatty acids (31% vs 51%), PUFAS (33% vs 13%), carotenoids (358 vs 169 mg kg1 dw) and for Ca (9 vs 19 g kg1 ), Fe (0.6 vs 4.2 g kg1 ), Cu (44 vs 14 mg kg1 ), Zn (93 vs 17 mg kg1 ) and As (2 vs 9 mg kg1 ). The chemical composition of U. clathrata indicates that it has a good potential for its use in human and animal food.

Liver injury from ampicillin-induced intestinal microbiota distresses in rats fed carbohydrate- and protein-rich diets

Purpose: To investigate the effect of ampicillin on rat intestinal microflora and liver in the presence of high carbohydrate and protein diets. Methods: Male Wistar albino rats were divided into four groups. The first group served as the control, the second group was treated with ampicillin (50 mg/kg for 3 weeks) and fed with a standard diet, while the third and fourth groups were treated with the same dose of ampicillin and fed with acarbohydrateand protein-rich diets, respectively, to observe the effect of diet on gut flora and liver. Fecal specimens were collected and used for qualitative determination of gut microbiota composition. Serum hepatospecific markers (AST, ALT and ALP) were estimated. The antioxidant status of liver tissues was estimated for GSH, MDA, GST, LDH and vitamin C l, in addition to sodium and potassium. Results: Administration of orogastric dose of ampicillin for 3 weeks induced inhibition of E.coli, yeasts, total anaerobes, and anaerobic lactobacilli with new growth of P. vulgaris and K. pneumonia. The levels of serum AST, ALT and ALP showed significant (p ˂ 0.05) increase to 163, 112.38 and 115.35 %, respectively in ampicillin-treated animals, compared to control. Also significant (p ˂ 0.05) increase in lipid peroxidation (120 %) and LDH (111 %) coupled with significant (p ˂ 0.05) decrease in glutathione (74.57 %), vitamin C (63.49 %) and glutathione S-transferase (41.51 %) were observed in ampicillintreated groups. No significant variation (p ˂ 0.05) in sodium and potassium levels were found between control and the treated group after 3 weeks of treatment. Conclusion: These results confirm that extended ampicillin therapy disrupts gut flora, which results in liver injury; hence, overuse of antibiotics should be avoid.