Determination of antibiotic resistance pattern and virulence genes in Escherichia coli isolated from bovine with subclinical mastitis in southwest of Iran

The aims of the present study were to investigate the prevalence of some virulence genes and also determine the antimicrobial resistance pattern of E. coli isolated from bovine with subclinical mastitis. The milk of 502 cows was collected from 8 dairy herds in the southwest of Iran. Conventional biochemical tests were used for identification of E. coli at the species level. Antimicrobial susceptibility patterns of E. coli isolates were determined by disc agar diffusion method and polymerase chain reaction (PCR) was used for detection of seven virulence genes including f17A, afaE-8, afaD-8, eaeA, cnf1, cnf2, and iucD. Seventy (13.94%) isolates of E. coli were identified in 502 milk samples. The highest rate of resistance was observed against tetracycline (18.6%), while none of the isolates were resistant to streptomycin. Eight (11.5%) out of 70 E. coli isolates carried at least one of the virulence genes. The afaD-8 was the most prevalent gene detected in 5 (7.1%) isolates. The afaE-8, iucD, and eaeA were detected in 3, 3, and 2 isolates respectively. Low prevalence of virulence factors may be indicating that most of the E. coli isolates originated from the commensal flora of cows and enter to the udders via environment contamination with feces.

     

Effects of probiotic lactic acid bacteria on growth performance,carcass characteristics,hematological indices,humoral immunity,and IGF-I gene expression in broiler chicken

The effects of screened lactic acid bacteria strains were evaluated on growth performance, humoral immunity, and IGF-1 gene expression in broiler chickens. The three dietary groups of negative control fed basal diet, the native LAB probiotic group (NP), and PrimaLac commercial LAB probiotic (PC) were studied. The results revealed that NP and PC diets significantly improved feed conversion ratio and increased body weight, as well as relative weight of carcass compared with group fed NC diet (P?<?0.05). Lymphocyte level was significantly increased in birds fed NP and PC (P?<?0.01), while serum triglycerides and total cholesterol levels were significantly decreased compared with the NC (P?<?0.05). Significant increases were observed in antibody titers against Newcastle disease virus of vaccinated birds (P?<?0.03), and morphological analysis of ileum revealed significant increases (P?<?0.05) in the villus height and villus height/crypt depth in birds fed NP and PC compared with the NC. The dietary significantly increased Lactobacillus spp. (P?<?0.05), while Escherichia coli (P?<?0.04) populations were significantly decreased, and also, the expression of IGF-1 gene in liver tissue of broilers fed NP and PC was significantly increased compared with the NC (P?<?0.05). These results indicated that the identified native LAB strains can be used commercially as a low-cost probiotic in poultry industry of Iran.

     

Influence of Stocking Density on Growth and Physiological Responses of Beluga,Huso huso (Brandt, 1869), and Ship Sturgeon,Acipenser nudiventris (Lovetsky, 1828), Juveniles in a Flow‐through System

The influence of three different initial stocking densities (low stocking density [LSD] = 1.5 kg/m²; medium stocking density [MSD] = 3 kg/m²; and high stocking density [HSD] = 6 kg/m²) in flow‐through systems was evaluated on growth and welfare in beluga, Huso huso, and ship sturgeon, Acipenser nudiventris, juveniles for 2 mo. Fish were kept in 18 concrete square tanks (2.0 × 1.0 × 1.2 m³) at 22.3 ± 0.4 C and under a natural photoperiod. In both species, the growth performance in terms of final body weight, weight gain, specific growth rate, and feed intake significantly decreased with increasing stocking density (P < 0.05). In both species, the percent of neutrophils increased after 60 d of trial (P < 0.05). Moreover, hematocrit and white blood cell counts increased after 60 d of trial in ship sturgeon (P < 0.05). Plasma immunoglobulin significantly decreased with increasing stocking density in both species. Plasma insulin‐like growth factor I decreased with increasing stocking density in beluga; however, it was not affected in ship sturgeon. Overall, these results showed that the LSD group in both species demonstrated more homogeneous and higher growth rate than the MSD and HSD groups.     

An in vitro comparative study of follicle stimulating hormone (FSH) and activin A effects on the maturation of preantral follicle-enclosed oocytes from immature Syrian mice

Objectives: It was aimed to investigate the effects of different doses of follicle stimulating hormone (FSH) and activin A on the growth and maturation of preantral mouse follicles during the in vitro culture. Methods: Preantral follicles (90-100 mum in diameter) were harvested from 6-8 week-old Syrian mice and cultured in TCM199 culture medium for 6 days to see the effect of FSH and Activin A. Activin A concentrations in the range of 10-200 ng/ml were used, while 10, 50, 100, 150 and 200 mIU/ml FSH were used in the experiment. Results: Activin A concentration of 100 ng/ml resulted in a significant increase in follicle diameter (170 mum) with the survival rate of 73% as compared to the control (100 mum and 25%, P<0.05). The number of oocytes matured and the percentage of germinal vesicle breakdown (GVBD) was 61 and 70%, respectively as compared to the control (20 and 29%, P<0.05). Follicle diameter (190 mum) and survival rate (85%) increased significantly in the presence of 100 mIU/ml of FSH as compared to the control (P<0.05). But, the administration of activin A+FSH increased the effect of both factors on follicular diameter (205 mum as compared to 100 mum in control, P<0.01). Follicle survival, oocyte maturation and GVBD rates were 91, 81 and 89%, respectively (P<0.01). Conclusion: These results have suggested that exposure to FSH and activin A before the formation of antral cavity had positive effect on follicle survival and oocyte robustness.     

Comparison of distribution of virulence determinants in clinical and environmental isolates of Vibrio cholera

Background: The virulence of a pathogenic Vibrio cholerae is dependent on a discrete set of genetic determinants. In this study, we determined the distribution of virulence determinants among the clinical and environmental isolates of V. cholerae. Methods: The antibiotic resistance profiles of the isolates were determined using standard disk diffusion assay. PCR assay was performed to analyze the presence of toxin genes of ctx, zot and ace. The composition of cholera toxin encoding element (CTX) region flanking of the V. cholerae isolates was also analyzed. Results: All of the clinical isolates (100%) showed a complete set of virulence genes and also the attachment site of the filamentous bacteriophage CTXphi. None of the environmental isolates contained the virulence genes and the attachment site of the CTXphi. Analysis of the flanking regions including the toxin-linked cryptic element and repeat in toxin genes revealed their integrity in the clinical isolates while in the environmental isolates they were absent or contained incomplete sequences. Comparison of the antibiotic resistance assay of the environmental and clinical isolates showed a significant difference in the resistance profiles of the isolates obtained from the two sites. High rates of resistance to co-trimoxosol, streptomycin and chloramphenicol were found with clinical isolates. Conclusion: The absence of all virulence determinants in the environmental strains may suggest that certain ecological features must be present for V. cholerae to acquire a complete set of virulence determinants and to turn them into pathogenic strains.     

Rapid screening of toxigenic vibrio cholerae O1 strains from south Iran by PCR-ELISA

BACKGROUND: The ability to sensitively detect Vibrio cholera with PCR-ELISA method represents a considerable advancement over alternative more time-consuming methods for detection of this pathogen. The aim of this research is to evaluate the suitability of a PCR-enzyme-linked immunosorbent assay for sensitive and rapid detection of V. cholera O1. METHODS: The 398-bp sequence of a gene that codes for the cholera toxin B subunit was amplified by PCR. The digoxigenin-labeled amplified products were coated on microplates and detected by ELISA. The PCR product was also hybridized with biotin labelled probe and detected by ELISA using streptavidin. RESULTS AND CONCLUSION: The specificity of the PCR was determined using 10 bacterial strains and 50 samples from south Iran. The detection limit was 0.5 pg of the genomic DNA and five bacterial cells. Adaptation of PCR into PCR-ELISA assay format facilitates specific and sensitive detection and diagnosis of human cholera disease. We conclude that this PCR-ELISA is a diagnostic method that specifically detects toxin genes in V. cholera O1 strains. It is more rapid and less cumbersome than other diagnostic methods for detection of toxicity in these strains.     

Physical soil properties and slope treatments effects on hydraulic excavator productivity for forest road construction

Effects of moisture, porosity and soil bulk density properties, grubbing time and terrain side slopes on pc 220 komatsu hydraulic excavator productivity were investigated in Miana forests road construction project which located in the northern forest of Iran. Soil moisture and porosity determined by samples were taken from undisturbed soil. The elements of daily works were measured with a digital stop watch and video camera in 14 observations (days). The road length and cross section profiles after each 20 m were selected to estimate earthworks volume. Results showed that the mean production rates for the pc 220 komatsu excavators were 60.13 m3 h(-1) and earthwork 14.76 m h(-1) when the mean depth of excavation or cutting was 4.27 m3 m(-1), respectively. There was no significant effects (p = 0.5288) from the slope classes' treatments on productivity, whereas grubbing time, soil moisture, bulk density and porosity had significantly affected on excavator earthworks volume (p < 0.0001). Clear difference was showed between the earthwork length by slope classes (p = 0.0060). Grubbing time (p = 0.2180), soil moisture (p = 0.1622), bulk density (p = 0.2490) and porosity (p = 0.2159) had no significant effect on the excavator earthworks length.     

Trophic status and primary production in Lake Choghakhor, Chaharmahal-Bakhtiyari Province, Islamic Republic of Iran

In this study, trophic state indicators, total phosphate, algal chlorophyll and Secchi disk transparency and zooplankton community of Lake Choghakhor was studied monthly between May 2003 and April 2004. This lake is a shallow ecologically and economically important water body in eastern part of Iran. Crop farming and recreational activities are examples of the human impact around and within the lake, leading to a loading of DIN (Dissolved Inorganic Nitrogen) and TP (Total Phosphate) into the lake. Now submerged plants especially Myriophyllum spicatum has covered almost the entire lake and dense macrophyte beds (Polygonom amphibium), located on the East Southern end of the lake appear to act as a sink for these nutrients. Lake Choghakhor appeared to be in a macrophyte dominated clear water state with low TP (annual mean: 24 +/- 15 microg L(-1)) and chlorophyll a (annual mean: 3 +/- 1.28 microg L(-1)) concentrations and very high Secchi depth. The grazing pressure of dominant pelagic filtering zooplankton Daphnia longespina did not seem to be significant in determining the low phytoplankton crop expressed as chlorophyll a. We expect that sequestering of nutrients by submerged plants and associated epiphytes are the dominant stabilizing mechanisms suppressing the phytoplankton crop of Lake Choghakhor.     

Study of water stress effects in different growth stages on yield and yield components of different rice (Oryza sativa L.) cultivars

A field experiment was conducted during 2001-2003 to evaluate the effect of water stress on the yield and yield components of four rice cultivars commonly grown in Mazandaran province, Iran. In northern Iran irrigated lowland rice usually experiences water deficit during the growing season include of land preparation time, planting, tillering stage, flowering and grain filing period. Recently drought affected 20 of 28 provinces in Iran; with the southeastern, central and eastern parts of the country being most severely affected. The local and improved cultivars used were Tarom, Khazar, Fajr and Nemat. The different water stress conditions were water stress during vegetative, flowering and grain filling stages and well watered was the control. Water stress at vegetative stage significantly reduced plant height of all cultivars. Water stress at flowering stage had a greater grain yield reduction than water stress at other times. The reduction of grain yield largely resulted from the reduction in fertile panicle and filled grain percentage. Water deficit during vegetative, flowering and grain filling stages reduced mean grain yield by 21, 50 and 21% on average in comparison to control respectively. The yield advantage of two semidwarf varieties, Fajr and Nemat, were not maintained under drought stress. Total biomass, harvest index, plant height, filled grain, unfilled grain and 1000 grain weight were reduced under water stress in all cultivars. Water stress at vegetative stage effectively reduced total biomass due to decrease of photosynthesis rate and dry matter accumulation.