Quantitative detection of the root-lesion nematode, Pratylenchus penetrans, using qPCR

Pratylenchus penetrans is one of the most economically damaging plant-parasitic nematodes and is found on a wide variety of crops. Correct identification and quantification of this nematode are necessary for providing advice to farmers, but are not easily obtained with the traditional way of microscopic observation. We developed a qPCR assay to detect and quantify P. penetrans in a short but accurate manner. A qPCR primer set, including two primers and a TaqMan probe, was designed based on the sequence of the β-1,4-endoglucanase gene. The assay was optimized by using the primers in a qPCR assay with SYBR green I dye and setting the qPCR program to different annealing temperatures ranging from 60 °C to 64 °C. Based on the Ct-values, we retained the program with an annealing temperature of 63 °C. The assay with the probe was very sensitive as it was able to detect a single individual of P. penetrans, even when mixed with up to 80 individuals of P. thornei. The specificity of the reaction was confirmed by the lack of amplification of DNA from 28 populations of 18 other Pratylenchus species and from plant-parasitic nematodes from nine other genera. DNA from 21 different isolates from P. penetrans was amplified. DNA extraction from 80 individuals and quantification by qPCR was repeated four times; Ct-values showed consistent results (Ct?=?24.4?±?0.4). A dilution series from DNA of P. penetrans resulted in a standard curve showing a highly significant linearity between the Ct-values and the dilution rates (R²?=?0.99; slope?=??3.23; E?=?104 %). The tests showed a high correlation between the real numbers of nematodes and the numbers detected by the qPCR. The developed qPCR assay provides a sensitive means for the rapid detection and reliable quantification of individuals of this pest. This method does not require expertise in nematode taxonomy and morphology, and can be used as a rapid diagnostic tool in research, as well as in diagnostic labs and extension services advising farmers for pest management.     

Effects of dietary barley inclusion and glucanase supplementation on the production performance,egg quality and digestive functions in laying ducks

This study evaluated the effects of barley inclusion and glucanase supplementation on the productive performance and digestive function in laying ducks. The experiment used a randomized design with a 5 × 2 factorial arrangement of 5 graded levels of barley (0%, 15%, 30%, 45% and 60%) with or without 1.5 g/kg β-1,3-1,4-glucanase (15,000 U/kg). During the experimental period of 120 d, the weight and total number of eggs within each pen were recorded daily, and egg quality was determined every 4 wk. At the end of the experiment, 3 randomly selected ducks within each replicate were sacrificed, then duodenal digesta and jejunal mucosa was collected. Dietary inclusion of barley had no effects on egg production, daily egg mass or FCR, but supplementation with glucanase improved egg production and FCR (P < 0.01). Barley did not affect feed intake of laying ducks, but glucanase tended to increase feed intake (P = 0.09). Neither barley nor β-glucanase had effects on the egg quality variables, except for yolk color score, which was decreased with increasing barley supplementation. Glucanase, but not barley, increased the activity of chymotrypsin and amylase in duodenal digesta. Barley inclusion affected the activity of alkaline phosphatase and maltase in jejunal mucosa (P < 0.05), but β-glucanase had no effects on the activity of these brush border enzymes. Barley inclusion increased the glucan content in duodenal digesta, but supplementation of glucanase to barley-based diet reduced digesta glucan content and reduced total volatile fatty acids and increased the proportion of acetic acid in cecal contents. The results indicate that, without glucanase, the optimal dietary barley level in the diets of laying ducks is about 13% for maximal production performance; glucanase supplementation of the barley diets improved production performance, probably through enhancing digestive function.     

Tryptophan in poultry nutrition: Impacts and mechanisms of action

Many studies have shown that productivity, immune system, antioxidant status, and meat and egg quality can be optimized by dietary supplementation with amino acids that are not usually added to poultry diets. Understanding the effects of these amino acids may encourage feed manufacturers and poultry producers to include them as additives. One of these amino acids is tryptophan (Trp). The importance of Trp is directly related to its role in protein anabolism and indirectly related to its metabolites such as serotonin and melatonin. Thus, Trp could affect the secretion of hormones, development of immune organs, meat and egg production, and meat and egg quality in poultry raised under controlled or stressed conditions. Therefore, this review discusses the main roles of Trp in poultry production and its mode (s) of action in order to help poultry producers decide whether they need to add Trp to poultry diets. Further areas of research are also identified to address information gaps.     

Study on prevalence and bacterial etiology of mastitis,and effects of subclinical mastitis and stage of lactation on SCC in dairy goats in Egypt

Tropical Animal Health and Production - In Egypt, inadequate information on prevalence and epidemiology of caprine mastitis is available. This study was designed to investigate prevalence and...     

Breaking seed dormancy in Cupressus atlantica Gaussen, an endemic and threatened coniferous tree in Morocco

Cupressus atlantica Gaussen (Cupressaceae) is an endemic and endangered coniferous tree geographically restricted to the N’Fis valley in South-Western Morocco. Like many forest species, C. atlantica exhibits dormancy which delays and reduces germination. To improveseed germination, different pre-treatments were conducted on C.atlantica seeds after storage for different periods (one, two and five years) including: scarification with sandy paper; soaking seeds in hot distilled water at 60℃and 80℃for 15 min and soaking seeds for 48 h in agibberellic acid (GA3) at 1,000 and 2,000 mg·L-1. Results showed that scarification with sandy paper increased the germination rate of Atlas cypress by up to 67%, indicating that the species possess essentially anexogenous dormancy (physical dormancy) due to the hard seed coat(hardseededness). Exogenous application of gibberellic acid (GA3) at1,000 mg·L-1 was also effective in breaking seed dormancy and germination induction. These two treatments induced faster speed germination expressed by low number of days to first germination (8 10days) and low values of mean germination times (MGT). However,germination rate, under any treatment, is greatly dependent on the year of seed collection. Seeds collected in year 2004 gave the highest value,suggesting that even after five years of storage, the germination capacity of C. atlantica seeds could remain high. This observation is very interesting in the exsitu conservation of such endemic and endangered species where the production of seeds is irregular over the years.     

Occurrence and distribution of ‘Candidatus Phytoplasma trifolii’ associated with diseases of solanaceous crops in Lebanon

A survey for phytoplasma diseases in tomato and pepper fields in Lebanon was conducted during 2003 and 2004. Tomato plants with stunting, yellowing or purplish leaves, proliferation of laterals buds, hypertrophic calyxes and virescent flowers were found in 25% of the tomato fields surveyed, where they represented 2–8% of the plants. Pepper plants displaying stunting and yellowing of leaves, were found in 27% of the fields and 1–4% of the plants were affected. Phytoplasmas infecting tomato and pepper had identical 16S-rDNA RFLP profiles and sequences. A phytoplasma isolate named PTL was transmitted by dodder from a diseased tomato plant to a periwinkle (Catharanthus roseus) plant in which it induced leaf yellowing, virescence and phyllody. 16S-rDNA phylogenetic analysis classified PTL as a strain of ‘Candidatus Phytoplasma trifolii’.     

Effect of egg parasitoid density on the population suppression of Sunn Pest, Eurygaster integriceps (Hemiptera: Scutelleridae), and its resulting impact on bread wheat grain quality

The impact of the density of Sunn Pest egg parasitoids, Trissolcus grandis Thomson (Hymenoptera: Scelionidae), at three levels of Sunn Pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), infestation was assessed on bread wheat variety Cham 6 in screened cages in the field for percentage of egg parasitism and grain quality. Percent parasitism ranged from 60.1 to 83.9%. No statistically significant differences were detected at parasitoid densities of 1, 2, or 3 insects per square meter at Sunn Pest densities of 2 or 4 insects per square meter. The quality of grain collected from all Sunn Pest infested cages without egg parasitoids was unacceptable for baking purposes and was significantly lower than grain from both non-infested plots and plots where either two or three egg parasitoids were present. The abundance of egg parasitoids in Al Hassakeh province in Syria was also determined. Egg parasitism was observed 1 week after the initiation of Sunn Pest oviposition and ranged between 22 and 59% in non-insecticide treated fields, while no parasitoids were recovered from insecticide treated areas within the same region. The impact of feeding by Sunn Pest on different wheat stages indicated that only feeding prior to the spike stage does not affect the bread-making quality of the grain.