Trace elements accumulation in soil and rice plants irrigated with the contaminated water

We carried out a study to see the effect of contaminated water of Nullah Dek on fine rice paddy and straw yields and trace elements accumulation in different parts of rice plants and soil. A site was selected near the bank of Nullah Dek at Kot Pindi Das in the District of Sheikhupura, Pakistan. The water of this nullah is contaminated by industrial effluents carrying different micronutrients. This water was employed to grow rice crop. Water samples were collected before transplanting and during the season with 15 days interval for analysis from 20 July to 1 November 2002 from a spot near village Shamke. Three fine rice varieties, viz. Super Basmati, Shaheen Basmati and Basmati 2000 were transplanted. These rice varieties were grown up to maturity. Paddy and straw yields data were recorded. Six composite soil samples from three random spots were collected from the experimental site before the start of the study to see the status of trace elements in soil. After the harvest of rice crop, soil, paddy and straw samples were analysed for Zn, Cu, Fe and Mn. The chemical analysis of Nullah Dek water showed that total salts concentration was greater than the safe limit, i.e. electric conductance (EC) > 1.0 dS m⁻¹. Even sodium adsorption ratio (SAR) was very high, but there was no problem of high residual sodium carbonate (RSC). Zn, Cu, Fe and Mn were present but within safe limits. The water of Nullah Dek remained within permissible limits of irrigation from onset of rainy season till 15 October. There was an increase in EC, SAR and trace elements concentrations after 15 October but within safe limits. Soil analysis revealed its saline nature, devoid of sodicity. Among trace elements, the zinc ranged between deficiency (<0.5 mg kg⁻¹) and adequate limits (>1.0 mg kg⁻¹). Copper, Mn and Fe were present in adequate amounts. After the harvest of rice crop there was a slight decrease in pH, EC_e and SAR at both the depths, while the concentrations of all trace elements were slightly increased with more in upper layer than the lower layer. Shaheen Basmati produced the maximum paddy yield followed by Basmati 2000 and then Super Basmati. The chemical analysis of paddy samples indicated a sufficient accumulation of zinc (1.68–1.78 mg kg⁻¹), copper (1.38–1.45 mg kg⁻¹), iron (6.12–6.37 mg kg⁻¹) and manganese (2.22–2.42 mg kg⁻¹). Analysis of rice straw also showed sufficient accumulation of zinc (27.50–28.50 mg kg⁻¹), copper (20.0–20.50 mg kg⁻¹), iron (270–280 mg kg⁻¹) and manganese (2.38–2.41 mg kg⁻¹).     

Influence of Plant Growth Promoting Rhizobacteria,Compost, and Biochar of Azolla on Rosemary (Rosmarinus Officinalis L.) Growth and Some Soil Quality Indicators in a Calcareous Soil

In order to study the effect of plant growth promoting rhizobacteria (PGPR), Azolla compost and Azolla biochar on some soil quality indicators and rosemary growth, a greenhouse experiment was conducted in a completely randomized design with six replications. Treatments consisted of T₁ (control), T₂ (1% Azolla compost), T₃ (1% Azolla biochar), T₄ (PGPR (P. fluorescens), T₅ (1% compost + PGPR) and T₆ (1% biochar + PGPR). Rosemary growth parameters and nutrients concentration increased in all treatments compared to control. Treatments increased soil nutrient concentrations, soil microbial respiration (SMR) and microbial biomass C (MBC) but decreased soil metabolic quotient (qCO₂) compared to control treatment. A significant enhancement in rosemary growth occurred due to the improved soil quality as a result of organic fertilizers application, particularly by co-application of P. fluorescens and compost or biochar of Azolla.     

Oocyte maturation,embryo development and gene expression following two different methods of bovine cumulus-oocyte complexes vitrification

Objective

To examine the maturational competence, embryo development and expression of genes involved in oocyte maturation and cumulus expansion (GDF9, BMP15, HAS2, TNFAIP6, FGF17 and FSHr) following two standard methods of bovine COCs vitrification.

Methods

Bovine cumulus-oocyte complexes (COCs) were aspirated from slaughtered ovaries and then distributed into three groups: non-vitrified COCs (control), vitrification 1 group (V1); vitrification was performed by 15% ethylene glycol (EG) and 15% DMSO in holding media (TCM-199 with 20% FCS); and vitrification 2 group (V2); vitrification was performed by 40% EG in holding media. After vitrification, COCs were warmed in two steps and cultured and then evaluated for nuclear maturation, embryo development and gene expressions.

Results

The mean (±SD) percentages of nuclear maturation and blastocyst/cleaved were higher in control group (79.5 ± 8.0 and 31.0 ± 5.1%) than the V1 (34.8 ± 9.1 and 4.4 ± 5.1%) and V2 (47.8 ± 11.7 and 7.1 ± 5.8%) groups (P < 0.05), respectively. Further, COCs in V2 group showed higher mean (±SD) percentages of cleavage compared to V1 group (31.8 ± 1.0 vs 21.7 ± 2.8%; P < 0.05). GDF9 and BMP15 expression levels were higher in COCs in the control than of the vitrification groups (P < 0.05). In addition, expression level of GDF9 and BMP15 was higher in V2 group than in V1group (P < 0.05). The expression of HAS2 and FGF17 in V1 group was lower (P < 0.05) than that of the V2 groups.

Conclusions

Expression of oocyte maturation genes was affected by vitrification procedure and conditions. Using EG alone for vitrification of bovine immature COCs, resulted in higher expression of GDF9, BMP15 and production of more in vitro matured and cleaved oocytes.

     

The effect of feeding rations with different ratios of concentrate to alfalfa hay on blood hematological and biochemical parameters of farmed ostriches (Struthio camelus)

A two-month trial was conducted to investigate the effects of pelleting rations with 70:30, 65:35, and 60:40 concentrate to high-quality roughage (alfalfa hay) ratios on hematological and biochemical parameters of ostriches. A total of 18 seven-month-old male ostrich chicks, initially weighing an average of 60–70 kg, were distributed into three different outdoor paddocks at a stocking density of six birds per paddock. In the morning, blood collection was made from the wing vein after about 12 h of fasting. All rations resulted in no significant change in hematological parameters. However, with increasing alfalfa inclusion rate, the heterophil/lymphocyte ratio tended to be lower (P?=?0.089); whereas the values of hemoglobin, packed cell volume (PCV), and mean corpuscular hemoglobin (MCH) tended to be higher (P values 0.069, 0.072 and 0.094, respectively). In terms of plasma biochemical parameters, increasing the ratio of concentrate to alfalfa hay in ostrich diets resulted in significant (P?<?0.05) depression in the values of glucose (up to 20.2 %), total cholesterol (up to 19.2 %), creatine kinase (up to 27.9 %), and aspartate aminotransferase (up to 29.9 %). Additionally, the 65:35 or 60:40 rations caused a decrease in low-density lipoprotein cholesterol level (27.9 and 42 %, respectively; P?=?0.008) compared to the 70:40 ration. Without exception, no rations affected the values of leukocyte subsets, total protein, albumin, globulin, uric acid, alanine aminotransferase, gamma glutamyl transferase, triglycerides, high-density lipoprotein cholesterol, and very low density lipoprotein. Based on our results, the concentrate to alfalfa ratio of 60:40 could be suggested as optimum ratio for good health conditions of juvenile ostriches.     

Humoral and cellular immune responses against Type I cysteine proteinase of Leishmania infantum are higher in asymptomatic than symptomatic dogs selected from a naturally infected population

Canids are natural reservoirs of Leishmania infantum and have been promoted as experimental hosts to decipher the pathogenesis of human visceral leishmaniasis (VL). In this study, the presence of IgG antibodies as well as the presence of mononuclear leukocytes reactive to different cysteine proteinases (CPs) were examined in 13 L. infantum-infected dogs (six with symptoms, seven asymptomatic). Cysteine proteinases which belong to papain-like enzymes known as clan CA are the most studied CPs of parasite protozoa. These molecules are expressed by the intracellular stages of the parasite and could be immunogenic. We studied Type II CP (CPA) and Type I CP (CPB) with its long C-terminal extension (CTE) which could be highly immunogenic. We showed that the level of antibodies reactive to rCPA is low in both symptomatic and asymptomatic dogs. In contrast, when CPB and CTE were used as antigens, the level of total IgG (with IgG2 superior to IgG1) reached higher values in asymptomatic dogs than in dogs with VL. While the peripheral blood mononuclear cell (PBMC) reactivity was significant when cultured in the presence of freezed/thawed (F/T) lysate, it remained low in presence of CP although always higher for PBMC recovered from asymptomatic dogs. We showed the importance of CPB and CTE in particular as a target of immune response and their potential use for serodiagnosis in asymptomatic dogs.     

Epidemiological aspects of canine visceral leishmaniosis in the Islamic Republic of Iran

An epidemiological study to examine the sero-prevalence of zoonotic visceral leishmaniosis (ZVL) among domestic and wild canines in endemic foci of Iran was carried out during 1999-2003 to assess the distribution of the disease and the possible association between infection in dogs, wild canines and people. Anti-leishmanial antibodies were detected by the direct agglutination test (DAT). Parasitological study was performed for all captured wild canines and were detected in some of the seropositive dogs with specific clinical signs (n=107). Serum samples (n=1568) were collected from domestic dogs in villages that are known endemic foci of human visceral leishmaniosis (HVL). Wild canine sera were collected from jackals (Canis aureus, n=10), foxes (Vulpes vulpes, n=10) and wolves (Canis lupus, n=10). Of the 1568 serum sampled collected from domestic dogs, 222 (14.2%) were positive by DAT (1:320 and above). No statistically significant difference was found between male (15.2%) and female (11.8%) sero-prevalence (P=0.083). Dogs of 8 years and above showed the highest sero-prevalence (40.6%). Only 23.9% of the seropositive domestic dogs had clinical signs. Parasitology and serology tests that were performed in 30 wild canines showed 10% these animals were infected by Leishmania infantum. Ten out of 11 Leishmania spp. isolated from the dogs and wild canines were identified as L. infantum and one other as L. tropica by molecular and biochemical techniques. For the first time in Iran, L. infantum and L. tropica were isolated from viscera of both a wolf and a domestic dog.     

Phylogenetic and Molecular Characterization of the Equine Influenza Virus A (H3N8) Causing the 1997 and 2004 Outbreaks in Morocco

Reported here are the results of antigenic and genetic characterization of equine influenza strains causing local outbreaks reported in Morocco, respectively, in 1997 and 2004. The antigenic and genetic characterizations of the equine influenza virus H3N8 are reported here. The highest similarity between the HA1 nucleotide sequences of A/equine/Nador/1/1997 and those of A/equine/Rome/5/1991 and A/equine/Italy/1199/1992 demonstrate that A/equine/Nador/1/1997 belongs to the European lineage. On the other hand, A/equine/Essaouira/2/2004 and A/equine/Essaouira/3/2004 were classified in the predivergent lineage. The present work emphasizes the importance of a national influenza survey program, which requires a collaborative laboratory network to promote the collection and characterization (antigenic and genetic) of equine influenza viruses in real time.     

Isolation by phage display of recombinant antibodies able to block adherence of Escherichia coli mediated by the K99 colonisation factor

K99 fimbriae are important for intestinal colonisation by bovine strains of enterotoxigenic Escherichia coli. The mode of action of this colonisation factor is well understood and specific immune responses are protective. K99 was therefore chosen for this study as a model to test if antibodies with anti-adhesion activity could be isolated from recombinant libraries using phage display techniques. Potentially, this strategy could be used to understand better the action of bacterial colonisation factors and aid the design of therapies (e.g. vaccines, purified protein products or bacteria bearing colonisation-blocking antibodies) to inhibit bacterial adherence. The major fimbrial subunit from K99, FanC, was purified from a clinical E. coli isolate. The protein was coated to plastic immunotubes and used as a target for selection of antibodies from the Tomlinson I and J libraries of single chain (scFv) antibodies. Clones able to recognise K99 were isolated by iterative rounds of binding, elution and amplification. scFv antibodies chosen from the resulting panel were purified and their specificity confirmed by ELISA. Pre-incubation of several scFvs with bacteria expressing K99 fimbriae inhibited the agglutination of erythrocytes. Further investigation by microscopy confirmed that when E. coli expressing K99 were exposed to scFv antibodies, the binding of bacteria to erythrocytes was blocked with high efficiency. The study showed that recombinant antibodies were able to block the action of a bacterial colonisation factor and hence that phage display techniques might be applied to the identification of less well-characterised virulence factors and the analysis of their structure and function.     

Changes of vitamins A, E, and C and lipid peroxidation status of breeding and pregnant sheep during dry seasons on medium-to-low quality forages

This study assessed the changes of plasma vitamin A, E, and C and the lipid peroxidation status of sheep during breeding and pregnancy under drought conditions. The study was conducted on 105 cross-bred fat tailed ewes, 3–5 years old with body condition scores (BCS) of 2.5 to 3.5. The ewes were grazing on medium-to-low quality forages during summer and low quality forages within the succeeding months and had ad libitum access to a mixture of alfalfa hay (40%) and wheat straw (60%) in the afternoons. From 3 weeks before breeding till 1 month after the introduction of rams, 300 g of barley grain/head/day was offered to the ewes and then the supplemental grain was reduced to 100 g/head/day. For better synchronization of estrus cycles in ewes, they were isolated from the rams for at least 2 months and then kept in close proximity of the rams for 1 week before the introduction of the rams to the ewe flock. Then, whole blood samples were collected on days 1, 7, 21, and 120 after ram introduction. Vitamins A, E, and C were measured in plasma. Malondialdehyde (MDA) was measured in the hemolysate as a biomarker of lipid peroxidation. Plasma progesterone (P4) was measured in the samples of day 120 for assessing pregnancy status of the ewes. Vitamins A and C showed continuous and significant declines (P < 0.05) through days 1 to 120. Vitamin E declined only during the first 21 days of the study and remained almost constant till day 120. MDA concentration increased significantly at day 21. An elevated concentration of MDA was also detected at day 120. The difference between days 21 and 120 was not significant (P > 0.05). A positive correlation between vitamins E and C was detected at day 120 (r = 0.349, P < 0.01). Age and BCS did not affect the patterns of changes. Assuming that the ewes with P4 concentrations ≥2.5 ng/ml were pregnant, 95 out of 105 ewes (90.5%) were pregnant at day 120 of the study. Under the conditions of the present study with medium-to-low quality pastures as the main sources of feed, ewes of various ages and body conditions may suffer from oxidative stress during breeding and pregnancy.