The effect of Froriepia subpinnata on the performance, carcass characteristics, blood parameters, immune system, microbial population, intestinal morphology, and breast meat fatty acid content of broiler chickens.

The present experiment was conducted to investigate the effects of F. subpinnata powder (FSP) on the performance, carcass characteristics, blood parameters, immune system, microbial population, intestinal morphology, and percentage of fatty acids in the breast meat of broiler chickens. A total of 300 day-old male chickens from the ROSS 308 strain were used in the form of a completely random design with four treatments of five replicates (15 birds each). The experimental treatments were, respectively, (1) control with base diet (without additives), (2) base diet + 1% FSP, (3) base diet + 2% FSP, and (4) base diet + 3% FSP. The results showed that the feed intake and weight gain increased in the treatments containing 2% and 3% FSP when compared to the control (P < 0.05). Cholesterol and ALT levels in the treatment containing 3% FSP were lower that the control (P < 0.05), while the concentration of glutathione peroxidase enzyme in the treatment containing 3% FSP significantly increased (P < 0.05). Thymus weight and antibodies produced against AIV in all three levels of FSP increased significantly compared to the control (P < 0.05). The population of lactobacilli and coliforms in the treatments containing FSP increased and decreased significantly compared to the control (P < 0.05). The length and width of the intestinal villi of the chickens that were fed with 3% of FSP had a significant increase compared to the control (P < 0.05). The percentage of saturated fatty acids in the breast decreased significantly with the consumption of all three levels of FSP (P < 0.001). In general, the results showed that the use of 3% FSP in the broiler diet increased the efficiency of growth performance and enzyme activity, while strengthening the immune system, favorably altering the intestinal microbial population, and reducing the fat in breast meat.

Dose-responses of zinc as zinc-methionine supplements on antioxidant status, hematological parameters, immune response and the expression of IL-4 and IL-6 genes of ewes in the hot season.

This study was implemented to evaluate the effects of different zinc doses as Zinc-Met supplement (Zinpro®) on the antioxidant status, blood immune cells, antibody titers, and the expression of IL-4 and IL-6 genes of ewes in the hot season. In a completely randomized design, 24 ewes were assigned to treatments as follow: 0, 15, 30 and 45 mg/kg zinc as Zinc-Met supplementation for 40 days in region with 40 °C and vaccinated against food-and-mouth disease as an immune challenge at day 30, and then blood samples were collected on day 40. Ewes were fed a basal diet containing 29.9 mg zinc/kg. The highest activity of the antioxidant enzyme and the lowest lipid peroxidation values were found in ewes receiving 30 and 45 mg/kg zinc following a linear trend. The highest lymphocytes count and antibody titers were found in ewes received 30 mg zinc/kg. There were no significant differences among treatments for the relative expression of genes. In overall, zinc supplementation non-significantly up-regulate interleukin-4 and down-regulate interleukin-6. It was concluded that zinc supplementation as Zinc-Met could enhance the antioxidant status and immune response of ewes under heat stress; supplementation of diet with 30 mg zinc/kg (300 mg/kg Zinpro®) appeared to be the most effective dose.

Effects of live and autolyzed yeast supplementation during transition period on ruminal fermentation, blood attributes, and immune response in dairy cows under heat stress condition.

This study was conducted to compare nutrient digestibility, performance and immune response of dairy cows received live and autolyzed yeast during the transition period in high ambient temperature. Cows (n = 25) were randomly divided and received a basal diet with or without live yeast or autolyzed yeast as on top three weeks pre-parturition until three weeks post-parturition. The Control group received a basal diet without yeast products; other groups received 0.5 g live yeast; 1.0 g live yeast; 10 g autolyzed yeast and 20 g/d/head autolyzed yeast. Live yeast resulted in higher nutrient digestibility compared with autolyzed yeast and the control. Methane production was the highest in autolyzed yeast and the lowest in live yeast. Average milk production was the highest in cows that received live yeast. The highest IgG level was for cows that received autolyzed yeast at a dose of 20 g/d/head. Live yeast had no significant effect, but autolyzed yeast increased the relative expression of γ-Interferon and interleukin-2 as compared with the control group. It was concluded that live yeast at a dose of 1.0 g/d/head could influence ruminal fermentation and milk production, but autolyzed yeast at a dose of 20 g/d/head could influence the immune response of dairy cows during the transition period and heat stress.

Inflammatory genes expression and calf responses to different dietary grains (steam flaked corn vs steam-rolled barley) and protein sources (pelleted soybean meal vs pelleted canola meal).

This study aimed to investigate the effects of grains, and protein sources in the starter diet on daily gain, pro- and anti-inflammatory genes expression, ruminal volatile fatty acid (VFA) concentration, and blood metabolites in the dairy calves. Forty Holstein calves were randomly assigned to treatments in a 2 × 2 factorial arrangement with the factors of grain source (steam-flaked corn, CG vs. steam-rolled barley, BG) and pelleted protein source (soybean meal, SBM vs. canola meal, CM). The daily gain of calves who fed with SBM was higher than those fed with CM. Total VFA concentration in the rumen of calves fed with CG was greater than BG at day 35 (p < 0.05), and calves receiving SBM had higher total VFA concentration than calves fed with CM. The relative expression of tumor necrosis factor-α and interleukin-1ß genes were significantly decreased in the calves fed with BG and CM compared to calves fed with CG and SBM. Calves receiving the starter diet based on BG had the highest (p < 0.05) gene expression of interferon-γ. Feeding calves with SBM is recommended because it resulted in a greater daily gain than CM. Complete replacement of SBM with CM, and CG with BG is not recommended in dairy calves.

The effects of thymol, oxalic acid (Api-Bioxal) and hops extract (Nose-Go) on viability, the Nosema sp. spore load and the expression of vg and sod-1 genes in infected honey bees.

This study was done to investigate the effects of thymol, fumagillin, oxalic acid (Api-Bioxal) and hops extract (Nose-Go) on Nosema sp. spore load, the expression of vitellogenin (vg) and superoxide-dismutase-1 (sod-1) genes and mortality of bees infected with N. ceranae. Five healthy colonies were assigned as the negative control, and 25 Nosema sp. infected colonies were assigned to five treatment groups including: the positive control: no additive to sirup; fumagillin 26.4 mg/L, thymol 0.1 g/L, Api-Bioxal 0.64 g/L and Nose-Go 5.0 g/L sirup. The reduction in the number of Nosema sp. spores in fumagillin, thymol, Api-Bioxal and Nose-Go compared to the positive control was 54, 25, 30 and 58%, respectively. Nosema sp. infection in all infected groups increased (p < .05) Escherichia coli population compared to the negative control. Nose-Go had a negative effect on lactobacillus population compared to other substances. Nosema sp. infection decreased vg and sod-1 genes expression in all infected groups compared to the negative control. Fumagillin and Nose-Go increased the expression of vg gene, and Nose-Go and thymol increased the expression of sod-1 gene than the positive control. Nose-Go has the potential to treat nosemosis if the necessary lactobacillus population is provided in the gut.

Hematological parameters, antioxidant status, and gene expression of γ-INF and IL-1ß in vaccinated lambs fed different type of lipids.

This study was aimed to evaluate the effects of vegetable oils as calcium salt on immune responses and the expression of immune-related genes in vaccinated lambs. Twenty-four lambs (35 kg body weight, 6 months old) were assigned to four treatments with six replicates in a completely randomized design for 40 days. Four concentrates were formulated in which the calcium salts of palm oil, canola oil, corn oil, and flaxseed oil were used. On day 30 of the experiment, lambs were vaccinated by a dose of foot-and-mouth disease virus. The blood samples were collected from jugular vein 10 days after vaccination. The level of malondialdehyde and the activity of liver enzymes were the highest in lambs receiving corn oil and the lowest in lambs receiving flaxseed oil. The highest lymphocytes and the lowest neutrophil percentages were observed in lambs receiving flaxseed oil. There was a significant difference among treatments for the relative genes expression. Flaxseed oil significantly upregulated interferon-γ and corn oil upregulated interleukin-1ß. The highest titer against foot-and-mouth disease virus was related to lambs receiving flaxseed oil, and the lowest titer was related to lambs that received corn oil. Flaxseed oil had more beneficial effects on immune response than other oils.

Effect of palmitic, linoleic and α-linolenic acids on blood cells count and interleukin-4, 8 genes expression in lambs experimentally infected with foot and mouth disease virus.

The current research was conducted to study the effects of different saturated and poly-unsaturated fatty acid sources in diets on immune responses of lambs experimentally infected with foot and mouth disease virus. A total of fifteen Sangsari male lambs with an average body weight of 48 ± 2 kg and an average age of 8 ± 1 months were randomly allocated to one of three experimental treatments: (1) PalOil group received a calcium soap of palm oil fatty acids in the diet as source of palmitic acid (16:0), (2) SunOil group received a calcium soap of sunflower oil fatty acids in the diet as source of linoleic acid (n-6 18:2), and (3) LinOil group received a calcium soap of linseed oil fatty acids in the diet as source of α-linolenic acid (n-3 18:3). The greatestand the least expression of IL-4 mRNA were measured in SunOil and LinOil groups, respectively. Expression of IL-8 mRNA was greater in SunOil treatment when compared with PalOil and LinOil groups. In conclusion, the results of the study confirmed that inclusion of α-linolenic acid in virally infected lambs'diet results in lower expression of IL-4 and IL-8 mRNA.

Effects of enzymatically hydrolyzed yeast supplementation on blood attributes, antioxidant status and gene expression of cytokines in vaccinated dairy cows.

This study was done to assess the effects of hydrolyzed yeast on blood cells counts, antioxidant status and gene expression of pro-inflammatory and anti-inflammatory cytokines of vaccinated dairy cows. Twenty Holstein lactating dairy cows were randomly divided into four groups received basal diet without (control) and with 10, 20 and 30 g/d/head of hydrolyzed yeast. Cows were vaccinated against foot and mouth disease virus at day 10 and blood samples were taken 10 days after. Cows received hydrolyzed yeast at doses of 20 and 30 g/d/head had the lowest and those in the control group had the highest malondialdehyde concentration. Cows received hydrolyzed yeast at dose of 20 g/d/head had the highest lymphocyte and the lowest neutrophil/lymphocyte ratio. IL-1ß and TNF-α downregulated and INF-γ up-regulated in cows received different doses of hydrolyzed yeast. The lowest expression of IL-1ß and TNF-α and the highest expression of INF-γ was for cows received hydrolyzed yeast at dose of 20 g/d/head. The highest titers against foot and mouth disease virus were for cows received hydrolyzed yeast at dose of 20 g/d/head and the lowest titer was for the control group. Supplementation of 20 g/d/head hydrolyzed yeast recommended in vaccinated cows.

Forage inclusion in calf starter has the best outcome when it is supplemented since 21 days after birth in Holstein calves.

To evaluate the optimum age of straw addition to starter diet on growth of calves, 40 Holstein calves (36.7 ± 4.96 kg) were assigned at 3 days of age to 1 of 4 treatments in a completely randomized design. Dietary treatments (% of dry matter) were as follows: control starter without any wheat straw (WS) inclusion until weaning (WS0); 90% calf starter concentrate (CSC) + 10% chopped WS offered from day 3 of age to weaning (WS3); 90% CSC + 10% chopped WS offered from day 21 of age to weaning (WS21); and 90% CSC + 10% chopped WS offered from day 40 of age to weaning (WS40). After weaning at 60 days of age, all calves received the same diet until finished at day 72 of age. Solid feed intake was affected by treatments whereby either pre- or post-weaning starter intake was greater for calves fed WS (P < 0.01), and WS3 fed calves had lower starter intake compared to WS21 and WS40 (P < 0.01). Weaning weight was the greatest for WS21 treatment compared to other groups (P < 0.01), although all forage fed calves were heavier than WS0 (P < 0.01). Accordingly, average daily gain also showed a similar pattern (P < 0.01). Without any difference at post-weaning, pre-weaning glucose was lowest in WS0 calves (P < 0.01). There were no effects of treatment on number of days that calves experienced a negative health criterion. In conclusion, we concluded that WS inclusion at d 21 of age is recommendable based on weaning and final BW.

Effects of different fat sources (saturated and unsaturated) on reproductive performance and biological indices of ewes during flushing period.

This study was conducted to evaluate the effect of feeding different sources of fat during flushing period on the reproductive performance, lambing percent, and twin numbers of Afshari ewes. A total of 84 ewes (mean weight 48 ± 3 kg; age: 3-4 years) were divided into seven groups of 12 animals and received flushing-specific rations for 5 weeks. The control group just received a basic ration (non-flushing). Lipid sources were calcium salt of palm oil (CaP), pure palm oil (PO), calcium salt of flaxseed (CaFL), calcium salt of sunflower oil (CaSF), flaxseed oil (FLO), and sunflower oil (SFO). Estrous cycles were synchronized in all ewes using 14-day CIDRs followed by 400-IU PMSG injection at the time of CIDR removal. Fertility and lambing percent were higher in ewes fed with diets containing calcium salts of flaxseed and SFO, as compared to other treatments. Total number of lambs in flushing treatments was significantly higher than that in the control group (p < 0.01). Serum cholesterol and progesterone levels were significantly greater in omega-3 (CaFL) and omega-6 (CaSF) treatments relative to other treatments (p < 0.01). It was concluded that supplementing the flushing diet with calcium salts of fatty acids (CSFA) increased blood metabolites and hormones related to reproductive performance; and improved fertility, lambing rate and ewes of CaFL treatment have the highest number of lambs (16 lambs) between different groups. Using saturated and unsaturated fatty acids, especially in their CSFA forms during flushing period, could improve the reproduction problems induced by progesterone deficiency, lack of durability of the fetus due to hormonal instability, and abortion control factors.

Isolation and characterization of Clostridium difficile in farm animals from slaughterhouse to retail stage in Isfahan, Iran.

To determine the prevalence of Clostridium difficile in farm animals from slaughterhouse through to retail stage, a total of 750 samples of feces, posteviscerated and washed carcass were collected from cattle, camels, goats, and sheep in Isfahan, Iran. The overall prevalence of C. difficile in feces, posteviscerated and washed carcass were 20 (13.3%), 23 (15.3%), and 11 (7.3%), respectively; while C. difficile was isolated from 79 (26.3%) retail samples. Twenty-nine (3.8%) isolates were toxigenic, with most toxigenic isolates (n = 17, 5.6%) identified from the retail stage. All toxigenic isolates harbored tcdA and tcdB; however, all were negative for cdtB. The 29 isolates were classified into 21 different ribotypes. This study revealed evidence of existence of toxigenic C. difficile in farm animal feces and meat in Iran.

Occurrence of Clostridium difficile in seasoned hamburgers and seven processing plants in Iran.

BACKGROUND: The recent increment of the incidence of Community Associated Clostridium difficile Infection (CA)-CDI has led to speculation that this disease is associated to foodborne transmission. Therefore it is critical to establish the community sources of CDI in order to implement the appropriate interventions. The present study was conducted to evaluate the prevalence of C. difficile in seasoned hamburger and examine the sources of C. difficile dispersal in hamburger processing plants. A total of 211 samples including hamburger ingredients, the final product, processing equipment and food contact surfaces were collected from seven hamburger processing plants to evaluate the routes of dispersal of C. difficile. The samples were assessed for the occurrence of C. difficile using culture and polymerase chain reaction (PCR) methods. All isolates were screened for the existence of toxin A, B and binary toxin genes. In addition, isolates were subjected to PCR ribotyping. RESULTS: Overall, 9/211 (4.2%) samples were positive. Toxigenic C. difficile were detected from 2/7 (28.5%) hamburger processing plants, in (3/54) 5.6% of beef meat samples, (2/56) 3.5% of swabs taken from the environment and (4/56) 7.1% of hamburger samples after both molding and freezing. C. difficile was not found in 45 non-meat ingredients including 14 defrosted onions, 14 textured soy proteins and 17 seasonings. All isolates contained tcdB gene while 7 strains were positive for tcdA and two remaining strains were negative for tcdA. None of the isolates harbored binary toxin gene (cdtB). PCR ribotyping of 9 isolates categorized into four ribotypes (IR21, IR 22, IR 23 and IR24). Ribotype IR 22 was the most common type 6/9 (66.6%) found. This genotype was isolated from raw meat, environmental samples and hamburger after both forming and freezing in one processing plant, suggesting raw beef meat as a possible major source of contamination. CONCLUSIONS: Hyper-virulent strains of ribotype were not found in this study however, occurrence of other toxicgenic strains indicate the public health significance of contamination of this product.

Effect of Organic Selenium Supplementation on the Antioxidant Status, Immune Response, and the Relative Expression of IL-2 and IFN-γ Genes in Ewes During the Hot Season.

This study was conducted to evaluate the effects of different doses of selenium (Se) from Sel-Plex© (selenium-enriched Saccharomyces cerevisiae yeast) supplement on the antioxidant status, the antibody titers against the foot-and-mouth disease virus, and the expression of interleukin-2 (IL-2) and interferon-γ (IFN-γ) genes in ewes during the hot season. Six ewes were kept at 25 °C and received basal diet (the negative control group), and 24 ewes were kept at 38 °C for 5 h per day and received no supplement (the positive control), 0.15, 0.30, and 0.45 mg Se/kg. Ewes in the positive control had higher (P<0.001) liver enzyme activity, malondialdehyde (MDA), and cortisol levels, and lower antibody titer than the negative control. The liver enzymes' lowest (P<0.001) activities were observed in ewes receiving 0.30 and 0.45 mg Se/kg. Ewes receiving 0.30 and 0.45 mg Se/kg had lower MDA levels than other treatments. Ewes receiving 0.30 and 0.45 mg Se/kg had higher (P<0.001) total antioxidant capacity levels than those receiving 0.15 mg Se/kg and the positive control. Se-supplemented groups had lower (P<0.001) relative expression of IL-2 and higher (P<0.04) expression of IFN-γ than the positive control. The antibody titer was the same in the positive control and the group receiving 0.15 mg Se/kg. Ewes fed a diet with 0.30 and 0.45 mg Se/kg had higher (P<0.011) antibody titer than the positive control. The Se supplementation can reverse the decrease of antioxidant capacity and immune function caused by heat stress, and 0.3 mg Se/kg from Sel-Plex©is the best dose.

Effects of different lipid sources with or without a probiotic on gastrointestinal tract, immune system and blood parameters of chickens: An animal model.

The objective of this study was to determine the effects of different lipid sources, with or without a probiotic, on the gastrointestinal tract, immune system and blood parameters of Ross 308 male chickens. In this study, 360 one-day-old chickens were randomly allotted to six treatments with six replicates. Experimental diets were: (1) control (CTL); (2) a diet containing 30 g/kg lipid from tallow (CTL+TLW); (3) a diet containing 30 g/kg lipid from soybean oil (CTL+SO); (4) the basal diet plus a probiotic (CTL+PRO), (5) a diet containing 30 g/kg tallow plus probiotic (TLW+PRO); and (6) a diet containing 30 g/kg soybean oil plus probiotic (SO+PRO). The percentage of liver and jejunum in the treatments that used tallow alone or tallow with probiotics had a significant increase as compared to the control. The villus height and crypt depth of the ileum and villus height/crypt depth in the treatments that used soybean oil and probiotic alone had a significant increase compared to the control. The weight of the spleen, bursa of Fabricius, and thymus in the treatments that used probiotics had a significant increase compared to the control. The amount of alkaline phosphatase and alanine aminotransferase as well as triacylglycerol in the treatment containing probiotic and its mixture with soybean oil had the least significant difference with the control. The results showed that the use of soybean oil, probiotics, and their mixture can improve intestinal morphology, strengthen the immune system, and reduce liver enzymes in chickens.

A new gene tree algorithm employing DNA sequences of bovine genome using discrete Fourier transformation.

Within the realms of human thoughts on nature, Fourier analysis is considered as one of the greatest ideas currently put forwarded. The Fourier transform shows that any periodic function can be rewritten as the sum of sinusoidal functions. Having a Fourier transform view on real-world problems like the DNA sequence of genes, would make things intuitively simple to understand in comparison with their initial formal domain view. In this study we used discrete Fourier transform (DFT) on DNA sequences of a set of genes in the bovine genome known to govern milk production, in order to develop a new gene clustering algorithm. The implementation of this algorithm is very user-friendly and requires only simple routine mathematical operations. By transforming the configuration of gene sequences into frequency domain, we sought to elucidate important features and reveal hidden gene properties. This is biologically appealing since no information is lost via this transformation and we are therefore not reducing the number of degrees of freedom. The results from different clustering methods were integrated using evidence accumulation algorithms to provide in insilico validation of our results. We propose using candidate gene sequences accompanied by other genes of biologically unknown function. These will then be assigned some degree of relevant annotation by using our proposed algorithm. Current knowledge in biological gene clustering investigation is also lacking, and so DFT-based methods will help shine a light on use of these algorithms for biological insight.

The effect of multiplex-PCR-assessed major pathogens causing subclinical mastitis on somatic cell profiles.

The major pathogens causing mastitis were evaluated by multiplex-polymerase chain reaction (M-PCR) with self-designed primers in four quarters of the first, third, and fifth parities in industrial, semi-industrial, and traditional dairy cattle farms in Iran. With the incidence of infection in the quarters by Staphylococcus aureus and Streptococcus agalactiae, the mean log somatic cell count (log SCC) increased from 5.06 to 5.77. The smallest changes occurred with Escherichia coli. Contagious pathogens, when compared with environmental pathogens, were more prevalent and common and created more profound quantitative and qualitative changes in SCC profiles. The second part of the study surveyed the diversity of contaminating pathogens and their effect on quantitative and qualitative profiles of somatic cells. M-PCR was used to determine the absence (M-PCR(-)) and presence of one (M-PCR(+1)), two (M-PCR(+2)), and three (M-PCR(+3)) major pathogens in raw milk samples. Quarter log SCC increased from 5.06 (for M-PCR(-1)) to 5.5 (for M-PCR(+1)), 5.7 (for M-PCR(+2)), and 6 (for M-PCR(+3)). Percent changes in polymorphonuclears (PMNs) were not significant between different quarters and parities but were significant between different farms in terms of pathogen diversity (P < 0.05). Therefore, by increasing the number of types of major pathogens involved in subclinical mastitis, SCC of udder quarters and the proportion of PMNs significantly increased, whereas the proportion of lymphocytes significantly decreased. This subject is very important in increasing the shelf life of dairy products, because PMNs are introduced to the enzymatic pools.

Functional Quails Eggs using Enriched Spirulina during the Biosorption Process.

Spirulina platensis was included at 2.5%, 5%, and 7.5% of the diet as a phytobiotic either as a crude preparation or after enrichment by a biosorption process and fed to 126 Japanese quail and the eggs collected and examined for 6 weeks. Assessments were made of physical and chemical characteristics of the eggs. All treatments with added Spirulina increased unsaturated fatty acids and decreased saturated fatty acids with the largest responses for linolenic (omega 3) and oleic (omega 9) acids. The changes in fatty acids were greater with enriched than crude Spirulina. These results suggest that eggs from quail fed with Spirulina may have positive effects on human health.

Effects of Supplemental Chromium Nanoparticles on IFN-γ expression of Heat Stress Broilers.

The aim of present study was to investigate the beneficial effect of chromium (III) picolinate (CrPic) and chromium (III) picolinate nanoparticles (NCrPic) addition on growth performance, stress-related hormonal changes, and serum levels of various immunity biomarkers, as well as the gene expression of IFN-γ in broilers exposed to heat stress conditions. Treatments included T1 which received the basal diet with no feed additive; T2 exposed to heat stress; T3, T4, and T5 containing 500, 1000, and 1500 ppb CrPic; as well as T6, T7, and T8 containing 500, 1000, and 1500 ppb NCrPic, respectively. After 2 weeks from CrPic and NCrPic supplementation, IFN-γ mRNA expression was assayed using the RT-PCR technique. The results showed that the lower body weight, daily weight gain, daily feed intake by heat stress, and the feed conversion ratio were recovered remarkably by CrPic and NCrPic supplements. The stress-elevated levels of cortisol and immunoglobulin were reduced significantly using CrPic and NCrPic supplementation (P ≤ 0.05). The gene expression profile showed that the upregulated expression of IFN-γ was regulated by the addition of CrPic and NCrPic, in particular, to the diet; however, a full downregulation of IFN-γ expression was observed after week 2 of NCrPic supplementation. In conclusion, the results indicated that nanoparticle supplementation could be effective in reducing heat stress-induced detrimental alterations, thereby attributing to substantial changes to the immune system, including IFN-γ expression.

Method validation for aflatoxin M1 determination in yoghurt using immunoaffinity column clean-up prior to high-performance liquid chromatography.

Yoghurt is a popular dairy product in Iran because of its beneficial influence on human health and nutritional value. Aflatoxin M1 (AFM1) is the metabolite of potential carcinogen aflatoxin B1, which can contaminate milk through the feed and is not eliminated by common processing heat treatment. An analytical method using immunoaffinity column for extraction and a high-performance liquid chromatography (HPLC) for quantification was developed for AFM1 in this study. An HPLC method with fluorimetric detection for the determination of AFM1 in yoghurt milk has been optimized and validated according to Commission Decision BS EN ISO 14501: 2007 by using the conventional validation approach. The procedure for determining selectivity, recovery, precision, decision limit (CCα) and detection capability (CCß) of the method has been reported. The results of the validation process demonstrate the agreement of the method with the provisions of Commission Regulation 401: 2006:EC. A new HPLC method with fluorescence detection was developed to determine aflatoxin M1. The detection limit was 1 ng/kg for yoghurt. The calibration curve was linear from 0.1 to 3.0 µg l⁻¹ injected. The method includes a preliminary clean-up and the average recoveries determined on three different days at the concentration levels of 0.025, 0.050 and 0.075 µg kg⁻¹ were in the range of 72.57%-86.66% with RSD in the range of 2.56%-8.41%. The interday and interlevel mean recovery value, which has been used to correct routine analysis results, was 80%. The method is rapid, easily automatable and therefore useful for accurate and precise screening of aflatoxin M1 in yoghurt.

Therapeutic effects of organic zinc on reproductive hormones, insulin resistance and mTOR expression, as a novel component, in a rat model of Polycystic ovary syndrome.

OBJECTIVES: Zinc is an effective factor in the reproductive system. Insulin resistance (IR) is known as an important disorder in patients with polycystic ovary syndrome (PCOS). Mammalian target of rapamycin (mTOR), which controls key cell activities, in particular, is activated in disorders such as PCOS. The present study was conducted to observe the therapeutic effects of organic zinc on IR, mTOR gene expression, and pathogenesis of PCOS in a rat model induced-PCOS. MATERIALS AND METHODS: Experimental treatments were performed on control and treated groups, consisting of healthy controls (Control, water, and standard feed intake and daily injection of sesame oil alone), Polycystic control (PCO, injection of 4 mg/kg estradiol valerate (EV) for four weeks). Treated groups (PCO-ZM 25, PCO-ZM 75, and PCO-ZM 175) after 4 weeks of receiving EV, were daily given three levels of 25, 75, and 175 mg zinc methionine/kg BW for 15 days, respectively. RESULTS: Injection of EV dramatically increased body and ovarian weights, levels of LH, testosterone, estradiol, triglyceride, fasting insulin, fasting glucose, HOMA-IR, IGF-1, gene expression of mTOR, and number of cysts (P<0.05). It also reduced the level of progesterone, HDL-C, and the number of antral follicles (P<0.05). However, by increasing zinc-methionine application especially at 175 mg/kg BW, the induction effects of EV were improved on ovarian cysts (P<0.05). CONCLUSION: Organic zinc showed beneficial effects in the EV induced PCOS rats via decreased insulin resistance and mTOR expression, restored the hormonal profile, and decreased the number of cysts in the ovaries.