Ultrastructural and molecular characteristics of Setaria species based on sequence analysis of genomic and mitochondrial gene markers in cattle (Bos taurus) and buffaloes (Bubalus bubalis) from Iran

ΔΕΝ ΔΙΑΤΙΘΕΤΑΙ ΠΕΡΙΛΗΨΗThe aim of the present study was to investigate the ultrastructural characteristics and genetic diversity of Setaria parasites from cattle (n=696) and buffalo (n= 522) from Khuzestan province of Iran and to compare them with available data from other countries/regions by sequences analysis of the 12S Rdna and the mitochondrial cytochrome C oxidase subunit I (cox1) genes. Based on SEM (Scanning Electron Micrographs) and light microscopy, all the isolated worms were identified as Setaria labiatopapillosa. Our results showed that 12.3% of cattle were infected with Setaria spp., while no infection was found in buffaloes. The maximal prevalence was observed in cattle younger than one year old. The prevalence rate was not influenced by the season of the year or gender. Comparison of the obtained sequences from Setaria with sequences of Setaria spp. from GenBank confirmed that all samples belong to the species S. labiatopapillosa. The phylogenetic tree constructed using cox1 and 12S rDNA genes of several other filarial nematodes showed that the Khuzestan isolates share a common branch with S. labiatopapillosa from other regions. Intra-specific variation was observed in 12S rDNA but not in cox1. In conclusion, our results indicating that S. labiatopapillosa is the main species involved in the spread of setarial infection in south-west of Iran and the identified worms corresponded mostly to worms that reported previously throughout other continents

Ocjena brzog imunoenzimnog testa u otkrivanju onečišćenja mlijeka, vode i izmeta bakterijom Salmonella Typhimurium

Serology-based detection methods have more applicable characteristics than standard bacterial culture methods. The aim of this study was to evaluate a cost-efficient, rapid, sensitive and simple test to diagnose milk, water and feces contaminated with Salmonella enteric subsp. enterica serovar Typhimurium (S. Typhimurium). The method developed was based on the addition of HRP-conjugated anti-Salmonella antibodies to the sample, precipitation of the complexes formed using centrifuge, and evaluation of the peroxidase activity of the sample supernatants. Raw cow’s milk, water and cattle feces samples were spiked with seven concentrations of S. Typhimurium. The optimized test recorded an accurate volume and concentration of HRP-conjugated antibodies, the bacteria, samples, ionic compounds and the precise time of the reaction. Also, the method of the test and standard sample preparation were carefully optimized. The tests were repeated at least ten times, and the minimum level of detectable bacteria was determined for each repetition. The results obtained were compared with the conventional bacterial culture method. The results showed that in an optimal condition, conjugated anti-Salmonella antibodies interacted with the bacteria in any matrix. The developed method can detect at least 30, 30, 3×103, and 3×103 cell/mL of S. Typhimurium in water, PBS, milk and feces samples respectively, in less than 20 minutes. Also, antibody detection using this EIA can detect an antibody concentration range of 4.3-70 μg/mL. The developed method is a rapid, cost-efficient and simple assay, and can be used to detect S. Typhimurium in various samples.Serološke dijagnostičke metode znatno su primjenjivije u odnosu na standardne bakteriološke kulture. Cilj ovoga istraživanja bio je ocijeniti ekonomsku isplativost, brzinu, osjetljivost i jednostavnost testa u otkrivanju onečišćenosti mlijeka, vode i izmeta bakterijom Salmonella enteric subsp. enterica serovar Typhimurium (S. Typhimurium). Metoda se temelji na dodatku HRP-konjugiranih anti-Salmonella protutijela u uzorku, precipitaciji nastalih kompleksa centrifugom i procjeni peroksidazne aktivnosti supernatanta. Uzorci sirovog kravljeg mlijeka, vode i izmeta goveda inokulirani su sa sedam koncentracija bakterije S. Typhimurium. Primjenom optimiziranog testa zabilježeni su precizni podaci o volumenu i koncentraciji HRP-konjugiranih protutijela, bakterija, uzoraka, ionskih spojeva i vremenu reakcije. Test i priprema standardnih uzoraka pažljivo su optimizirani. Test je ponovljen najmanje deset puta i u svakom je ponavljanju određena najmanja razina detektibilnih bakterija te su rezultati uspoređeni s konvencionalnim pretragama kultura. Rezultati su pokazali da su u optimalnim uvjetima konjugirana anti-Salmonella protutijela reagirala s bakterijama u svakom tipu uzoraka. Razvijena metoda može u manje od 20 minuta otkriti barem 30 stanica/mL bakterije S. Typhimurium u vodi, 30 stanica/mL u PBS-u, 3×103 stanica/mL u mlijeku i 3×103 stanica/ mL u izmetu. Također, primjena ove metode mogla bi otkriti koncentraciju protutijela od 4,3 do 70 μg/mL. Metoda je brza, jeftina i jednostavna te može otkriti bakteriju S. Typhimurium u različitim uzorcima

Axial Strength of Purlins Attached to Standing Seam Roof Panels

The purpose of this research was to determine the axial load capacity of Z-purlins with one flange attached to a standing seam roof system. The axial load capacity was determined by developing a relationship between the flexural uplift buckling strength and the axial buckling strength in the Z-purlin. This relationship was investigated using finite element models and by conducting a parametric study. At the conclusion of the parametric study, confirmatory tests were conducted to verify the finite element results. A relationship has been provided that relates the axial buckling strength in the Z-purlin to the flexural uplift buckling strength. This paper is a condensed version of the MBMA Research Report 99-02 (MBMA 2001)

Effect of Lactobacillus plantarum and Lactobacillus delbrueckii subsp. bulgaricus on growth performance, gut microbial flora and digestive enzymes activities in Tor grypus (Karaman, 1971)

In this study, two main probiotics (Lactobacillus plantarum and Lactobacillus delbrueckii subsp. bulgaricus) were isolated from the intestine of Tor grypus and their effects on growth performance, gut microbial flora and digestive enzymes activities (i.e., α-amylase, lipase, trypsin, chymotrypsin, and alkaline phosphatase) of Tor grypus (Karaman, 1971) were determined. Juveniles of T. grypus (n=480, weighing 45±10 g) were randomly divided into 4 equal groups (with 3 replications) and fed on diets containing 5×10^7 cfu g^-1 of L. plantarum (G1), L. bulgaricus (G2), L. casei (G3) and a control diet (without probiotic) for 60 continuous days. Results showed that probiotic supplementations had generally significant effects on growth performance (p<0.05). The G2 group had the best effect on specific growth rate (SGR) and feed utilization efficiency in T. grypus. The digestive enzyme activities, on day 30 after feeding, significantly increased in G1 when compared with those in the control group (p<0.05). Significant changes in bacterial intestinal flora were observed in all probiotic groups compared with the control (p<0.05). Our results indicated that supplementation of isolated bacteria from the intestine of T. grypus (i.e., G2) can efficiently improve growth performance, intestinal microbiota and some digestive enzyme activities in juvenile T. grypus. Therefore, it can be isolated and used as a growth enhancer just like the commercial probiotics

Formulation for multiple curved crack problem in a finite plate

The formulation for the curved crack in a finite plate is established. The technique is the curved crack in a finite plate is divided into two sub-problems i.e. the curved crack problem in an infinite plate and the finite plate without crack. For the first problem, the curved problem is formulated into Fredholm integral equation, where as for the second problem the complex boundary integral equations based on complex variables are considered. The solution of the coupled boundary integral equations gives the solution on the domain of the boundary

Investigation of Polar and Nonpolar Cyclotides Separation from Violet Extract Through Microfluidic Chip

Cyclotides (CTs) as a cyclic peptide obtained from different groups of plants have been very attractable field of research for scientists because of their specific properties like their natural function as host defense agents. CTs are bioactive peptides from plants that characterized by their head-to-tail cyclic backbone and knotted arrangement of their three conserved disulfide bonds. Their natural function is thought to be as host defense agents and a single plant can express dozens to hundreds of CTs. CTs stand out as a family of antimicrobial peptides (AMPs) because of their exceptional stability, structural plasticity, unique biochemical target, and Gram-negative selective antimicrobial action. These features together with recent advancements in the methods of production of CTs make them an intriguing prospect from a drug development perspective. To accomplish this aim, as part of a separation, detection and research of anti-cancer properties CTs study, we investigate the separation of cyclotides in violets into polar and non-polar groups by microfluidic chips

Immunohistochemical and bacteriological investigations of mannheimia haemolytica in sheep bronchopneumonia

Mannheimia haemolytica infection is one of the most common etiologic agents of sheep pneumonia almost all over the world. Ovine pneumonic Mannheimiosis is characterized by severe fibrinous pleuropneumonia. Subacute to chronic cases progress to purulent bronchopneumonia and its squeals include abscessation and fibrous pleural adhesions. In the present study, lungs of 8986 sheep were inspected grossly in the Ahvaz abattoir and totally 65 lungs with visible signs of bronchopneumonia were selected for pathological and bacteriological examinations. Mannheimia haemolytica antigens were detected in 63.07% of immunoperoxidase stained tissue sections while 52.30% of the lungs were positive in bacteriological culture. Suppurative, necrotic and fibrinous types of bronchopneumonia were the most abundant lesions and right cranial lobes, specifically their cranial portions, were the most affected areas. McNemar test showed a significant difference between the diagnostic power of immunohistochemistry (IHC) and bacterial culture in detection of M. haemolytica (κ=0.66). Considering IHC as a golden test, sensitivity and specificity of bacterial culture were estimated as 78.05 and 91.67%, respectively. Chi- squared test showed significant correlations between the distribution of the lesions and bacterial isolation (P=0.04), types of lesions and IHC results (P=0.01), and also types of bronchopneumonia and mixed/pure isolation (P=0.008). This study showed the significant role of Mannheimia haemolytica in causing pneumonic lesions of studied sheep

Estradiol attenuates spinal cord injury-induced pain by suppressing microglial activation in thalamic VPL nuclei of rats.

In our previous study we showed that central pain syndrome (CPS) induced by electrolytic injury caused in the unilateral spinothalamic tract (STT) is a concomitant of glial alteration at the site of injury. Here, we investigated the activity of glial cells in thalamic ventral posterolateral nuclei (VPL) and their contribution to CPS. We also examined whether post-injury administration of a pharmacological dose of estradiol can attenuate CPS and associated molecular changes. Based on the results,in the ipsilateral VPL the microglial phenotype switched o hyperactive mode and Iba1 expression was increased significantly on days 21 and 28 post-injury. The same feature was observed in contralateral VPL on day 28 (P<.05). These changes were strongly correlated with the onset of CPS (r(2)=0.670). STT injury did not induce significant astroglial response in both ipsilateral and contralateral VPL. Estradiol attenuated bilateral mechanical hypersensitivity 14 days after STT lesion (P<.05). Estradiol also suppressed microglial activation in the VPL. Taken together, these findings indicate that selective STT lesion induces bilateral microglia activation in VPL which might contribute to mechanical hypersensitivity. Furthermore, a pharmacological dose of estradiol reduces central pain possibly via suppression of glial activity in VPL region

Effect of microchannel dimensions in electrochemical impedance spectroscopy using gold microelectrode

Microfluidic chip systems have been an area of interest for lab-on-a-chip and organ-on-a-chip studies in recent years. These chips have many advantages such as high efficiency, low sample consumption, fast analysis, durability and low cost. Today, electrochemical sensors are frequently applied in microfluidic chips because of their potential for label-free detection and low-cost production. A commonly employed electrochemical technique is electrochemical impedance spectroscopy (EIS), which captures changes in phase and amplitude as signal passes through the system under test. In the utilization of microelectrodes within microfluidic channels, noise becomes a problem in EIS measurements. In this study, EIS measurements were performed using microfluidic chips with various dimensions of width while the properties and dimensions of the microelectrodes were kept constant. It was found that the results of cyclic voltammetry (CV) cleaning and EIS experiments deteriorated when smaller than 1 mm wide-microchannels were integrated onto 100 µm wide microelectrodes. These finding sets the basics for on-chip electrochemistry experiments using microfluidic integrated microelectrodes and therefore is fundamentally important in future on-chip EIS measurements