Comparative antifungal activities and biochemical effects of monoterpenes on plant pathogenic fungi

The antifungal activity of twelve monoterpenes, camphene, (R)-camphor, (R)-carvone, 1,8-cineole, cuminaldehyde, (S)-fenchone, geraniol, (S)-limonene, (R)-linalool, (1R,2S,5R)-menthol, myrcene and thymol was evaluated against four plant pathogenic fungi Rhizoctonia solani, Fusarium oxysporum, Penecillium digitatum and Asperigallus niger by using mycelial growth inhibitory technique. (S)-limonene and thymol were examined for their inhibitory effects on pectin methyl esterase (PME), cellulase and polyphenol oxidase (PPO) of tested fungi. Thymol was the most potent antifungal compound against the four test fungi with EC₅₀ values of 33.50, 50.35, 20.14 and 23.80 mg/L on R. solani, F. oxysporum, P. digitatum and A. niger, respectively. The antifungal activity of thymol was comparable to a reference fungicide, carbendazim. (S)-limonene and 1,8-cineole exhibited pronounced antifungal activity against the four tested fungi. The most effective antifungal compounds thymol and (S)-limonene showed strong inhibitory effect on the activity of PME and cellulase but revealed no inhibitory effect on PPO. The results showed that PME was more sensitive than cellulase to thymol and (S)-limonene. This is the first report on the inhibitory effects of monoterpenes thymol and (S)-limonene on PME, cellulase and PPO. The results indicated that monoterpenes may cause their antifungal activity by inhibiting PME and cellulase. The strong antifungal activity of thymol, (S)-limonene and 1,8-cineole reported in this study indicated that these compounds have a potential to be used as fungicides.     

Murine typhus in the homeless

Homeless populations are particularly exposed to many vector-borne diseases because of their poor living conditions. We tested sera from 299 homeless people recruited in 2010 and 2011 in Marseilles, France for antibodies to Rickettsia typhi by microimmunofluorescence using a titer of 1:25 as a cut-off titer, and we confirmed the results by Western blot and cross-adsorption studies. Sixty-three persons (22%) had antibodies against R. typhi. The murine typhus seroprevalence rates have significantly increased in homeless populations between the 2000-2003 and 2010-2011 periods. These findings indicate that the homeless are increasingly exposed to flea-borne murine typhus in Marseilles. One might suggest that multiple strikes of sanitation workers resulting in the increase of waste and construction sites combined with the poor living conditions of the homeless expose this population to rodents and their fleas. Further annual studies are necessary to follow rodent-associated diseases among homeless people in Marseille.     

Laparoscopic Anatomy of the Abdomen and Laparoscopic Ligating Loops,Electrocoagulation, and a Novel Modified Electroligation Ovariectomy in Standing Mare

The purpose of this study was to provide a detailed laparoscopic anatomy of the caudal abdominal region of mare in a standing position and to evaluate and modify a technique for standing laparoscopic ovariectomy using combination between hand-tied ligating loop and electrocoagulation techniques, as the ligating loops, electrocoagulation, and modified electroligation laparoscopic ovariectomy were applied using nine adult mares. Laparoscopy was practical and effective for direct visual examination of internal abdominal organs in the mare. Ventral dislocation of abdominal viscera after pneumoperitoneum was established with the mare in standing position, which provided an excellent inspection of the dorsal and ventral structures in the peritoneal cavity on the right and left sides. Standing laparoscopic ovariectomy using an electroligation modified method was considered a safe and effective method for hemostasis of the mesovarium, technically easy, time saving, and economical. The mean surgical time for bilateral ovariectomy was 40 ± 7.63, 60 ± 5.25, and 85 ± 6.43 minutes for electroligation-modified technique, ligating loops technique, and electrocoagulation technique, respectively.     

D-lactate-induced neurotoxicity in calves

D-lactic acidosis occurs as a complication of short-bowel syndrome in humans and in a variety of other gastrointestinal disorders in monogastrics and ruminants. It is associated with signs of impaired central nervous system (CNS) function including ataxia and coma. The objective of this experiment was to determine if either acidification of nervous tissue or D-lactic acid is responsible for decreased neurological function. Eight Holstein calves (32 ± 11 d) were surgically catheterized with indwelling intravenous jugular and atlanto-occipital space cerebrospinal fluid (CSF) catheters and infused for 6 h in random order with iso-molar DL-lactic acid (DL-LA), L-lactic acid (L-LA), hydrochloric acid (HCl) or saline. DL-LA induced ataxia after 4 h of infusion and produced the greatest obtunding of CNS function (at 7 h, score 8.0 ± 0.4), whereas, the other infusions caused neither ataxia nor scores over 1.5 (p < 0.01 from DL-LA). After 6 h, DL-LA induced significantly less acidemia than HCl (pH 7.13 ± 0.06 and 7.00 ± 0.04, base excess −16 ± 1 and −23 ± 3 mmol/l, bicarbonate 11 ± 1 and 8 ± 1 mmol/L respectively, all p < 0.01), but greater than L-LA and saline (p < 0.01). CSF changes followed a similar but less pronounced pattern. Although HCl infusion produced a severe acidemia and CSF acidosis, only minor effects on neurological function were evident suggesting that D-lactate has a direct neurotoxic effect that is independent of acidosis. Conversely, L-LA produced only minor neurological changes.     

Efficacy of nano-hydroxyapatite prepared by an aqueous solution combustion technique in healing bone defects of goat

The present study was undertaken to evaluate porous hydroxyapatite (HAp), the powder of which was prepared by a novel aqueous solution combustion technique, as a bone substitute in healing bone defects in vivo, as assessed by radiologic and histopathologic methods, oxytetracycline labeling, and angiogenic features in Bengal goat. Bone defects were created in the diaphysis of the radius and either not filled (group I) or filled with a HAp strut (group II). The radiologic study in group II showed the presence of unabsorbed implants which acted as a scaffold for new bone growth across the defect, and the quality of healing of the bone defect was almost indistinguishable from the control group, in which the defect was more or less similar, although the newly formed bony tissue was more organized when HAp was used. Histologic methods showed complete normal ossification with development of Haversian canals and well-defined osteoblasts at the periphery in group II, whereas the control group had moderate fibro-collagenization and an adequate amount of marrow material, fat cells, and blood vessels. An oxytetracycline labeling study showed moderate activity of new bone formation with crossing-over of new bone trabeculae along with the presence of resorption cavities in group II, whereas in the control group, the process of new bone formation was active from both ends and the defect site appeared as a homogenous non-fluoroscent area. Angiograms of the animals in the control group showed uniform angiogenesis in the defect site with establishment of trans-transplant angiogenesis, whereas in group II there was complete trans-transplant shunting of blood vessel communication. Porous HAp ceramic prepared by an aqueous combustion technique promoted bone formation over the defect, confirming their biologic osteoconductive property.     

Multilocus sequence analysis supports a low genetic diversity among ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma australasia’ related strains infecting vegetable crops and periwinkle in Egypt

‘Candidatus Phytoplasma australasia’ causes important damages to the Egyptian vegetable crop production. A prerequisite for controlling the different diseases it causes to eggplant, tomato and squash, is to trace its propagation pathways. To allow the differentiation of ‘Ca. P. australasia’ strains, a multilocus sequence analysis protocol was developed. Four conserved phytoplama genes namely tuf, secY, dnaK and dppA, were selected among the CDS of a ‘Ca. P. aurantifolia’ genome draft. The corresponding genes were PCR amplified from tomato, eggplant and squash collected in 2010 from the governorates Sharkia, Elmynia and Beni sueif, as well as from Catharanthus roseus periwinkles collected in 2013 from Kafrelsheikh governorate. Sequence comparisons showed no diversity among the Egyptian isolates of ‘Ca. P. australasia’ that also constitute a distinct cluster within the 16SrII-D taxonomic subgroup. This low diversity supports a common epidemiology for the different diseases affecting vegetable crops and periwinkle in Egypt and suggests that future investigations on insect vector should focus on polyphagous leafhoppers.     

Effect of Cinnamon (Cinnamomum zeylanicum) on Growth Performance,Feed Utilization,Whole-Body Composition,and Resistance to Aeromonas hydrophila in Nile Tilapia

Four isonitrogenous (30% crude protein) and isocaloric (4.40 kcal/g) experimental diets were formulated to contain 0.0% (control), 0.5%, 1.0%, or 1.5% cinnamon. The diet containing 1% cinnamon resulted in significantly greater (P < 0.05) specific growth rate (SGR), feed conversion ratio (FCR), feed efficiency ratio (FER), protein efficiency ratio (PER), apparent protein utilization (APU), and energy utilization (EU). In regard to body composition, there was no significant difference in dry matter, protein, lipids, and ash (P > 0.05) contents due to cinnamon supplementation. Blood plasma profile showed an improvement in hemoglobin (HB), red blood cell (RBCS), hematocrite (PCV), total protein, and total lipids, while there was a decrease in creatinin, urea, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and glucose in fish fed 1% cinnamon. Cinnamon was found to have an antibacterial activity antagonistic to Aeromonas hydrophila infection in fish. Performance and FCR improvement translated into a 10% decrease in feed costs associated with the 1% cinnamon diet.     

X-ray diffraction analysis of Yemeni cotton fibers

The lowland and coastal regions are the areas where cotton is cultivated in Yemen. The land used for this purpose exceeds 120 acres and expandable in the upcoming seasons. We have selected the earlier two varieties of cotton fibers cultivated in two different areas Abyan and Zabid. Wide Angle X-ray Scattering (WAXS) data from these fibers have been recorded and analysed to obtain the micro-structural parameters with the application of Line Profile Analysis (LPA). Linked Atom Least Squares (LALS) program has been used to obtain molecular structure and packing in these fibers. For the first time micro-structural parameters of these cotton fibers cultivated in Yemen are computed and reported. This study will be of help to understand the structure-property relation in these cotton fibers.     

Prevalence of brucellosis and infectious bovine rhinotracheitis in organized dairy farms in India

This study was carried out to investigate the prevalence of bovine brucellosis and infectious bovine rhinotracheitis (IBR) in organized dairy farms with history of abortion in India. ELISA and Rose Bengal Plate Test (RBPT) were used to detect the seropositive animals and the test results indicated that 22.18% and 13.78% animals were declared as sero-positive by ELISA and RBPT, respectively. Milk Ring Test (MRT) was carried out only in one farm and 12.82% of the tested animals were turned positive. Culture examination analysis of milk samples, two animals revealed the presence of organisms indistinguishable from Brucella spp. The organism was confirmed as brucella by morphological characteristics and biochemical tests. An overall sero-prevalence of antibodies against IBR was found to be 60.84%. None of the genital and nasal swab samples was found to be positive for presence of bovine herpesvirus -1 (BHV-1) on repeated passage in Madin-Darby Bovine Kidney (MDBK) cell lines. Brucella and IBR considered as the causal agent for abortions in these farms. The present study indicates the urgent need and the necessity for control of these infectious diseases which cause heavy economic losses to the organized farms.     

An efficient adventitious shoot regeneration system on excised leaves of micropropagated lingonberry (Vaccinium vitis-idaea L.)

Summary

An efficient system to regenerate shoots in vitro on excised leaves of lingonberry (Vaccinium vitis-idaea L.) was developed. Leaf explants from shoot-proliferating cultures produced multiple shoots without an intermediary callus phase on zeatin (ZN)-containing shoot induction media within 3–4 weeks of culture initiation. Cultivars Regal and Splendor, and one clone from a natural stand in Estonia (ECL1), were used in the first experiment. Young expanding leaves with the adaxial side touching the culture medium, and maintained for 7 d in darkness, produced the best results. There were significant genotypic differences in adventitious shoot formation. A second experiment studied the effects of ten concentrations of three cytokinins: ZN at 5, 10, 20, 30 and 40 μM; 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (thidiazuron, TDZ) at 0.1, 1, 5 and 10 μm; and 6-(γ-γ-dimethylallylamino) purine (2ip) at 25 μM were compared with leaf segments of different polarity in ‘ECL1’. Zeatin was found to be more effective than TDZ or 2iP as an inductive signal for regenerating many vigorous shoots. Zeatin induced multiple shoot formation at all concentrations tested, but maximum morphogenic response was observed at 20 to 30 μM. The media containing TDZ generally promoted more callus formation and suppresed shoot elongation. In a third experiment with the lingonberry cultivar Erntedank and the clone ‘ECL1’, a new medium developed for lingonberry shoot culture proved more effective than the modified Murashige and Skoog medium for regenerating shoots on leaf explants. Elongated shoots were excised and rooted directly on a 2 peat:1 perlite (v/v) medium after dipping in 0.8% indole-3-butyric acid. Rooted plantlets were acclimatized under greenhouse conditions to evaluate somaclonal variation.