Phylogenetic background and virulence genes of Escherichia coli isolates from colisepticemic and healthy broiler chickens in Iran

The purposes of this study were to determine the phylogenetic background and the virulence gene profiles of Escherichia coli isolates from colisepticemic and feces of healthy (AFEC) broiler chickens. In this study, 253 E. coli isolates including 141 avian pathogenic E. coli (APEC) and 112 AFEC isolates were examined by PCR. In general, 253 E. coli isolates distributed among group A (51.8%), B1 (15.8%), B2 (8.7%), and D (23.7%). Ten (8.9%) AFEC isolates segregated in to B1 phylo-group and 102 (91.1%) isolates fell into six different phylogenetic subgroups. Distribution of colisepticemic and fecal isolates differed significantly in their assignments to A and B1 phylo-groups. The three most prevalent virulence genes were crl, fimH, and aer in isolates between both groups. The four genetic markers aer, papC, afa, and sfa were detected significantly more often among colisepticemic isolates than in fecal isolates from healthy broilers. The presence of stx ₂ gene in fecal isolates were significantly differs among the colisepticemic isolates. F17 fimbrial family encoding gene and eae gene were detected in APEC and AFEC isolates, respectively. The colisepticemic and fecal isolates possessed the virulence genes were detected in all of the four phylogenetic groups. Several combination patterns of the virulence genes were detected in APEC and AFEC isolates. In colisepticemic isolates the combination of aer, crl, and fimH genes was the most prevalent pattern. None of the examined isolates harbored the cdt, cnf1, ipaH, and stx ₁ virulence gene sequences.     

Evaluation of genetic relationships among Iranian pistachios using microsatellite markers developed from Pistacia khinjuk Stocks

To evaluate the genetic relationships among wild and cultivated Pistacia species grown in Iran and the analysis of genetic variation among Iranian pistachio genotypes, two DNA libraries enriched for dinucleotide (AG)_n and trinucleotide (ATG)_n microsatellite motifs were developed from Pistacia khinjuk genome. Following screening of clones by colony PCR technique, 44 clones were sequenced and 27 pairs of primers designed from flanking regions of the repeats. The examination of primer pairs, designed from P. khinjuk sequences, showed successful cross-species amplification within the genus Pistacia. A dendrogram constructed on the basis of the Minimum Evolution clustering algorithm revealed that Pistacia vera has closer relationships with P. khinjuk, than with Pistacia integerrima, Pistacia palaestina, Pistacia atlantica and Pistacia mutica. The dendrogram further distinguished the wild Sarakhs pistachio from the rest of P. vera genotypes suggesting that the domesticated genotypes of P. vera are evolved from P. vera var. Sarakhs and then this wild genotype likely develops to other local pistachios. Hence, it seems that the wild Sarakhs pistachio plays an important role in evolutionary trend of the edible pistachios in Iran. The results indicated that microsatellites developed in P. khinjuk are distributed in the genome of indigenous pistachio species including P. vera genotypes and therefore they will be useful in characterization of Iranian pistachio genotypes.     

Statistical optimization of wool dyeing with Alizarin Red S as a natural dye via central composite design

Recently a revival interest in the use of natural dyes in textile coloration has been growing. The major parts of natural dyes are anthraquinones. Depending on substituents, anthraquinone compounds are dyes, as represented by Alizarin Red S (ARS) which is a major constituent of the natural colorant madder. In this study, colorization of wool fiber by ARS as a natural dye was studied. The progress of the colorization process was followed spectrophotometrically at 500 nm as λ _max. The effective factors on the process were investigated using one factor at a time (OFT) method. Then, the central composite design (CCD) method was applied to design experiments for the evaluation of the interactive effects of the four most important operating variables. The values of the optimized factors for OFT and CCD methods in wool dyeing with ARS were respectively as follow: temperature 70 °C and 55 °C, L/R: 40/1 and 40/2, pH 5.0 and 2.5 and time 60.0 min and 42.0 min. The predicted results by CCD had significantly higher exhaustion percentages and relatively better fastness properties than OFT and were found to be in a good agreement (R²=0.9908) with those obtained by performing experiments. Finally, the obtained results were shown a good wash fastness for dyed wool with ARS.     

Cover‐crop seeding‐date influence on fall nitrogen recovery

Planting cover crops after corn‐silage harvest could have a critical role in the recovery of residual N and N from fall‐applied manure, which would otherwise be lost to the environment. Experiments were conducted at the University of Massachusetts Research Farm during the 2004–2006 growing seasons. Treatments consisted of oat and winter rye cover crops, and no cover crop, and four cover‐crop dates of planting. The earliest planting dates of oat and winter rye produced the maximum biomass yield and resulted in the highest nitrate accumulation in both cover‐crop species. The average nitrate accumulation for the 3 years in winter rye and oat at the earliest time of planting was 60 and 48 kg ha^–1, respectively. In 2004 where the residual N level was high, winter rye accumulated 119 kg nitrate ha^–1. While initially soil N levels were relatively high in early September they were almost zero at all sampling depths in all plots with and without cover crops later in the fall before the ground was frozen. However, in plots with cover crops, nitrate was accumulated in the cover‐crop tissue, whereas in plots with no cover crop the nitrate was lost to the environment mainly through leaching. The seeding date of cover crops influenced the contribution of N available to the subsequent crop. Corn plants with no added fertilizer, yielded 41% and 34% more silage when planted after oat and rye, respectively, compared with the no–cover crop treatment. Corn‐silage yield decreased linearly when planting of cover crops was delayed from early September to early or mid‐October. Corn‐ear yield was influenced more than silage by the species of cover crop and planting date. Similar to corn silage, ear yield was higher when corn was planted after oat. This could be attributed in part to the winter‐kill of oat, giving it more time to decompose in the soil and subsequent greater release of N, while the rapidly increasing C : N ratio of rye can lessen availability to corn plants. Early plantings of cover crops increased corn‐ear yield up to 59% compared with corn‐ear yield planted after no cover crop.     

The effects of ketamine‐midazolam anesthesia on intraocular pressure in clinically normal dogs

Objective To determine the effects of intravenous ketamine‐midazolam anesthesia on intraocular pressure (IOP) in ocular normotensive dogs. Animals Thirteen adult mixed‐breed dogs. Procedures Dogs were randomly assigned to treatment (n = 7) and control (n = 6) groups. Dogs in the treatment group received intravenous ketamine 15 mg/kg and midazolam 0.2 mg/kg and dogs in the control group received intravenous saline. The time of intravenous drug injection was recorded (T₀). Measurements of IOP were then repeated 5 min (T₅) and 20 min (T₂₀) following the intravenous administration of ketamine‐midazolam combination and saline in both groups. Results Measurements showed normal IOP values in both groups. The mean ± SD baseline IOP values for treatment and control groups were 13.00 ± 1.47 and 10.33 ± 2.20, respectively. For baseline IOP values, there was no significant difference between treatment and control groups (P = 0.162). In the treatment group, the subsequent post‐treatment mean ± SD values were 15.64 ± 2.17 (5 min), and 14.92 ± 1.98 (20 min). There was no evidence of statistical difference between baseline values and post‐treatment values after treatment with ketamine‐midazolam (P₅ = 0.139; P₂₀ = 0.442). In control eyes, the mean ± SD values at 5 and 20 min were 10.41 ± 2.01 and 10.16 ± 1.69, respectively. There was no significant difference between baseline values and post‐treatment values in control group (P₅ = 1.000; P₂₀ = 1.000). Conclusion Ketamine‐midazolam combination has no clinically significant effect on IOP in the dog.     

Effects of wastewater application on saturated hydraulic conductivity of different soil textures

As metropolitan areas expand, the municipal and industrial uses of freshwater increase. Therefore, water resources for irrigation become limited and wastewater reuse for irrigation becomes a good alternative. For this purpose, the effects of suspended solids in wastewater on the soil physical properties, i.e., saturated hydraulic conductivity, K_s, have to be considered. The objectives of this research were to study the effects of applying freshwater and differently treated wastewater on K_s in the surface and subsurface layers of sandy‐loam, loam, and clay‐loam soils. This effect was studied by investigating the ratio of K_s for wastewater to K_s for fresh water in soil surface as K_r1 and in soil subsurface as K_r2. The results showed that the application of freshwater did not reduce the K_r1 considerably. However, the reduction in K_r1 mainly occurred in soil depth of 0–50 mm due to the application of wastewater. This effect is more pronounced in clay‐loam soil than in loam and sandy‐loam soils. It is concluded that application of wastewater with TSS (total suspended solid) of ≥ 40 mg L^–1 resulted in K_r1 reduction of >50% in different soil textures. However, the K_r2 reduction at soil depth of 100–300 mm is not considerable by application of wastewater for different soil textures. Further, it is concluded that less purified wastewater can be used in light‐texture soils resulting in less reduction in K_r1. Empirical models were developed for predicting the value of K_r1 as a function of amounts of wastewater application and TSS for different soil textures that can be used in management of wastewater application for preventing deterioration of soil hydraulic conductivity.     

A sensitive biosensor based on gold nanoparticles to detect <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> in soil

Ralstonia solanacearum, the devastating causal agent of potato bacterial wilt, is a soil-borne bacterium that can survive in the soil for a long time. The development of sensitive on-field detection methods for this pathogen is highly desirable due to its widespread host range and distribution. A novel nanobiosensor was thus developed to detect unamplified genomic DNA of R. solanacearum in farm soil. Gold nanoparticles functionalized with single-stranded oligonucleotides served as a probe to detect R. solanacearum genomic DNA. The advantages of this strategy include rapidity, facile usage and being a visual colorimetric method.     

Comparison of rapid diagnostic tests to detect <Emphasis Type="Italic">Mycobacterium avium</Emphasis> subsp. <Emphasis Type="Italic">paratuberculosis</Emphasis> disseminated infection in bovine liver

Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s disease, a chronic enteritis in cattle and other domestic and wild ruminants. The presence of MAP in tissues other than intestines and associated lymph nodes, such as meat and liver, is a potential public health concern. In the present study, the relationship between the results of rapid diagnostic tests of the Johne’s disease, such as serum ELISA, rectal scraping PCR, and acid-fast staining, and the presence of MAP in liver was evaluated. Blood, liver, and rectal scraping samples were collected from 200 slaughtered cattle with unknown Johne’s disease status. ELISA was performed to determine the MAP antibody activity in the serum. Acid-fast staining was performed on rectal scraping samples, and PCR was performed on rectal scraping and liver samples. PCR-positive liver samples were used for mycobacterial culture. Overall, the results of this study demonstrated that MAP can be detected and cultured from liver of slaughtered cattle and rapid diagnostic tests of Johne’s disease have limited value in detecting cattle with MAP infection in liver. These findings show that the presence of MAP in liver tissue may occur in cows with negative results for rapid diagnostic tests and vice versa. Hence, liver might represent another possible risk of human exposure to MAP. Given concerns about a potential zoonotic role for MAP, these results show the necessity to find new methods for detecting cattle with MAP disseminated infection.     

Results of Schirmer tear test in clinically normal llamas (Lama glama)

Purpose To determine the normal reference range for Schirmer tear test (STT) values in clinically normal llamas (Lama glama) Animals Nine captive llamas (Lama glama) (seven females and two males) were used in this study. Procedure Complete ophthalmic examinations were performed without chemical restraint. STT I values were evaluated in both eyes of all llamas using a commercial STT strip of a single lot number (Schirmer‐Tränentest^®, Germany). STT II value was also measured in both eyes of seven female llamas. Results No statistically significant differences among ages or between right and left eyes were found for any of the results. The mean ± SD STT I of 18 eyes of nine llamas was 17.3 ± 1.1 mm/min (Range 15–19 mm/min). The mean ± SD STT II of 14 eyes of seven llamas was 15.4 ± 1.7 mm/min (Range 12.5–17.5 mm/min). A paired samples t‐test demonstrated that there was a significant difference between the STT I and II values (P = 0.001). Conclusion This study provides novel data for normal reference ranges of STT I and II values in healthy llamas. Results of this study may assist veterinarians in the diagnosis of ocular surface disease and syndromes affecting the tear film in these species.