Expression of Toll-like receptors and downstream genes in lipopolysaccharide-induced porcine alveolar macrophages

The aim of the present study was to determine the age-related kinetic changes of Toll-like receptors (TLRs) and downstream genes expression, and secretion of cytokine in lipopolysaccharide (LPS) stimulated porcine alveolar macrophages (AM). For this purpose, AMs were isolated from 5-day-old newborn piglets and 120-day-old young pigs. mRNA expression and cytokine measurement was determined by quantitative real-time PCR and ELISA, respectively. First, AMs were incubated for 24 h in the absence or presence of increasing concentrations of LPS. Results showed the up-regulation of TLRs 2, 4, 5 and 9 mRNA from all concentrations of LPS used, as compared to non-stimulated cells, and TLR4 was the highest expression in both ages (P<0.05). Furthermore, quantitative analysis demonstrated increased expression of mRNAs encoding TLRs 2, 4, 5 and 9, LBP, CD14, MD2, MyD88, IRAK4 and TRAF6 in both ages in a time-dependant manner (P<0.05). Overall, LPS inducible mRNA for TLR4, LBP, CD14 and MyD88 had higher expression in newborn piglets compared with those of young pigs (P<0.05). The level of cytokine protein IL6 and TNFα in supernatant fluid significantly varied with time of incubation and age of animals. Their concentration increased immediately at 1 h after LPS stimulation and remained significantly higher up to 48 h in both ages. Production of pro-inflammatory cytokine protein IL6 and TNFα in supernatant was significantly higher in young pigs than those of piglets. This study suggests that differential age-related changes in the expression of TLRs and downstream genes, and pro-inflammatory cytokine could contribute to a different age-related innate immune response during pulmonary infection. Further investigation is warranted to determine the precise effects of LPS on porcine AMs by means of a functional study across a wider age range.     

Available (ileal digestible reactive) lysine in selected cereal-based food products

True ileal total lysine digestibility was determined and compared with true ileal reactive lysine digestibility when applied to 20 cereal-based breakfast foods. Semisynthetic diets each containing a breakfast cereal as the sole protein source were formulated and fed to growing rats. Titanium dioxide was included as an indigestible marker. Digesta were collected from the rats and total (conventional amino acid analysis) and reactive (guanidination) lysine were determined in both diets and digesta. The true ileal reactive lysine digestibility ranged from 53 to 108% and was significantly higher than the true ileal total lysine digestibility for most of the breakfast cereals. Available lysine content (digestible reactive lysine content) ranged from 0.21 to 3.5 g/kg across the breakfast cereals. The conventional measure of digestible total lysine content significantly overestimated (on average 37%) available lysine for the majority of the cereals. Breakfast cereals undergo a significant degree of lysine modification probably as a result of processing during manufacture.     

Over-mature beech trees (Fagus orientalis Lipsky) and close-to-nature forestry in northern Iran

Oriental Beech is the most important commercial tree species in northern Iran. In recent years wood production companies interested in felling large beech trees for profit have challenged advocates of close-to-nature silviculture who favor conservation. Our study objective was to assess the economic value of over-mature beech trees by relating tree diameter (DBH) to amount of decay. Based on the location of onset of decay, we categorized three types of decay as stump, stem, and crown decay. Trees of greater diameter (age) typically showed greater decay in the stem. Percent of decayed volume, diameter of decayed tissue, and length of decay in tree stems varied between 0.5%?64.3%, 15 cm?75 cm, and 2.0?19.5 m, respectively. With increasing trunk diameter, the proportion of truck decay increased. Red heart and dark red heart constituted 25% and 14.3% of sampled trees, respectively. However, we found no correlation between intensity of stem decay and morphological characteristics of trees. Seedlings were not abundant around the bases of over-mature trees, suggesting that the trees did not contribute to regeneration of the stand. Beech trees of diameter >1 m do not provide valuable round wood for industries and cause to raise wood production costs. We recommend that these trees >1 m DBH should be retained in forest stands because of their low commercial value but high ecological and conservational values such as maintaining biodiversity in forest ecosys-tems.     

Long-term effect of fire on herbaceous species diversity in oriental beech (Fagus orientalis Lipsky) forests in northern Iran

We studied the long-term impacts of natural burning on herbaceous species diversity 37 years after a fire occurred in the Roudbar forests of Guilan Province, northern Iran. Numerous studies have examined short-term changes in understory vegetation following wildfire; however, very few long-term studies are available or changes inferred from retrospective studies based on chronosequences. For this study, 170 ha of forest (85 ha, burned areas; 85 ha, unburned areas) were surveyed. Because the 1000 m 2 plots were too large for detailed measurements of herbaceous species, we determined a sub-sample size according to the Whittaker’s nested plot sampling protocol and minimal areas method. Hence, sub-plots of 32 m 2 were used for herbaceous species measurements, which consisted of percent cover of each species based on the Domin criterion. We measured plant diversity (Shannon-Wiener index), species richness (Margalef’s index), and evenness (Smith-Wilson index). Mean percent cover, together with diversity, richness and evenness, increased markedly in burned areas compared to unburned controls. This suggests that the biodiversity of these forests could be restored within 37 years after fire. However, the abundance of invasive species such as Rubus fruticosus and Bromus benekenii increased significantly in burned areas, but these could be controlled by relevant silvicultural operations.     

Evaluation of four different crops’ sensitivity to sulfosulfuron and tribenuron methyl soil residues

ABSTRACT

On the basis of greenhouse bioassays, the sensitivities of root and shoot biomass of barley (Hordeum vulgare L.), corn (Zea mays L.), oilseed rape (Brassica napus L.) and sugar beet (Beta vulgaris L.) to soil-incorporated sulfosulfuron and tribenuron methyl were assessed. Shoot and root biomass production was measured 30 days after emergence. Plant responses, including roots and shoots dry weight per pot, were described by a three parameter log-logistic regression model as a function of sulfosulfuron or tribenuron methyl doses and the relative sensitivities were calculated at the various ED-levels with their associated 95% confidence intervals. The most precise ED-levels were that at ED₅₀ and sensitivity rank was oilseed rape, sugar beet, corn and barley, whatever the chosen response level and herbicide. We calculated relative sensitivities at ED₁₀, ED₅₀ and ED₉₀ for the species of barley, the most tolerant crop, as reference. Comparison of relative sensitivity of crops to both herbicides showed that the sensitivity of these crops was much larger for tribenuron methyl than for sulfosulfuron. Oilseed rape was the most sensitive species (ED₅₀?=?0.202 and 0.179?µg?kg^?1 soil for root dry weight (RDW) response to sulfosulfuron and tribenuron methyl, respectively) while barley was the most tolerant one (ED₅₀?=?1.008 and 3.68?µg?kg^?1soil for RDW response to sulfosulfuron and tribenuron methyl, respectively). Sugar beet and corn had intermediate sensitivity. Also, we demonstrated how important it is to show the confidence intervals of relative sensitivities. In several instances the relative sensitivities, even numerically large, were not significantly different from 1.00. We demonstrate that classifying biotypes as resistant to a herbicide requires the threshold of resistance/susceptible (R/S) of 4.00 only be accepted if confidence intervals do not cover 1.00.     

Survey of the microbiological count of the milk collected from livestock ancient city of Garmsar,Semnan Province,Iran

Milk has a high nutritional value and can affect human health. Therefore, it is important to know the factors affecting its quality. The purpose of this study was to evaluate the raw milk microbial quality and factors affecting it. A total of 4260 raw cow milk samples, which were collected from villages around Garmsar city, Iran, and their microbial load were registered at Garmsar Dairy Cow Association between 2013 and 2014, was used in the present study. The total average ± SD of microbial load concentration was 3.02?×?10⁵?±?2.75?×?10⁵ (CFU/ml). The average concentration of total bacteria in 2013 was 2.73?×?10⁵?±?1.58?×?10⁵ (CFU/ml) and in 2014 it was 3.32?×?10⁵?±?3.93?×?10⁵(CFU/ml). Seasonally, it was 2.94?×?10⁵?±?1.72?×?10⁵ (CFU/ml), 3.69?×?10⁵?±?4.55?×?10⁵ (CFU/ml), 2.91?×?10⁵?±?1.62?×?10⁵ (CFU/ml), and 2.63?×?10⁵?±?1.67?×?10⁵ (CFU/ml) for spring, summer, autumn, and winter, respectively. There was a significant difference in seasonal average microbial concentration of milk samples (p?<?0.05). The present study concludes that microbial load of milk can be affected by environmental and hygienic conditions. Ranchers need to pay more attention to hygiene especially in warm seasons. Because in case of high microbial load in raw milk, even after milk pasteurization, it is not possible to have a high-quality and desirable product. Milk producers and distributors need also awareness in order to reduce the incidence of secondary contamination. But, the main correction is to establish direct supervision of the Ministry of Health onto the health of dairy cow farmers by structural integrity of Veterinary Medical Organization as the global one health.

     

Screening for probiotic characters in lactobacilli isolated from chickens revealed the intra-species diversity of Lactobacillus brevis

Considering the importance of the poultry industry and the increasing interest in alternative growth promoters, probiotics are considered as a potential candidate for use in the poultry industry. In this study, Lactobacillus species were isolated from 21 rectal swabs of 11 healthy 6-day-old and 10 healthy 21-day-old chickens and their fecal and feed samples. The isolates were characterized and their probiotic characteristics, including resistance to gastric acid and bile salts, biofilm formation and adherence to epithelium or mucus, amylase and protease activity and production of inhibitory compounds, were assessed. From 31 acid and bile resistant lactobacilli, only 2 Lactobacillus brevis and 1 Lactobacillus reuteri strains showed significant probiotic properties. These isolates indicated detectable attachment to Caco-2 cells and significant antibacterial activities against Gram-positive and Gram-negative pathogens. Additionally, phenotypic and genotypic diversity of lactobacilli isolates were studied by Phene Plate (PhP) system (PhP-LB) and random amplified polymorphic DNA (RAPD)-PCR, respectively. PhP-LB results of 24 L. brevis isolates showed a high phenotypic variation among the isolates. In comparison, results of RAPD-PCR highlighted a low diversity. Therefore, it seems that combination of the 2 techniques (PhP and RAPD-PCR) could result in a significant discriminatory power than each of them used alone.     

Carbon mineralization in subtropical alluvial arable soils amended with sugarcane bagasse and rice husk biochars

Subtropical recent alluvial soils are low in organic carbon (C). Thus, increasing organic C is a major challenge to sustain soil fertility. Biochar amendment could be an option as biochar is a C-rich pyrolyzed material, which is slowly decomposed in soil. We investigated C mineralization (CO₂-C evolution) in two types of soils (recent and old alluvial soils) amended with two feedstocks (sugarcane bagasse and rice husk) (1%, weight/weight), as well as their biochars and aged biochars under a controlled environment (25 ±2 ℃) over 85 d. For the recent alluvial soil (charland soil), the highest absolute cumulative CO₂-C evolution was observed in the sugarcane bagasse treatment (1 140 mg CO₂-C kg^-1 soil) followed by the rice husk treatment (1 090 mg CO₂-C kg^-1 soil); the lowest amount (150 mg CO₂-C kg^-1 soil) was observed in the aged rice husk biochar treatment. Similarly, for the old alluvial soil (farmland soil), the highest absolute cumulative CO₂-C evolution (1 290 mg CO₂-C kg^-1 soil) was observed in the sugarcane bagasse treatment and then in the rice husk treatment (1 270 mg CO₂-C kg^-1 soil); the lowest amount (200 mg CO₂-C kg^-1 soil) was in the aged rice husk biochar treatment. Aged sugarcane bagasse and rice husk biochar treatments reduced absolute cumulative CO₂-C evolution by 10% and 36%, respectively, compared with unamended recent alluvial soil, and by 10% and 18%, respectively, compared with unamended old alluvial soil. Both absolute and normalized C mineralization were similar between the sugarcane bagasse and rice husk treatments, between the biochar treatments, and between the aged biochar treatments. In both soils, the feedstock treatments resulted in the highest cumulative CO₂-C evolution, followed by the biochar treatments and then the aged biochar treatments. The absolute and normalized CO₂-C evolution and the mineralization rate constant of the stable C pool (K_s) were lower in the recent alluvial soil compared with those in the old alluvial soil. The biochars and aged biochars had a negative priming effect in both soils, but the effect was more prominent in the recent alluvial soil. These results would have good implications for improving organic matter content in organic C-poor alluvial soils.