Expression of chicken LEAP-2 in the reproductive organs and embryos and in response to <Emphasis Type="Italic">Salmonella enterica</Emphasis> infection

In recent years host antimicrobial peptides and proteins have been recognised as key mediators of the innate immune response in many vertebrate species, providing the first line of defense against potential pathogens. In chickens a number of cationic antimicrobial peptides have been recently identified. However, although these peptides have been studied extensively in the avian gastrointestinal tract, little is known about their function in the chicken reproductive organs and embryos. Chicken Liver Expressed Antimicrobial Peptide-2 (cLEAP-2) has been previously reported to function in protecting birds against microbial attack. The aim of this study was to investigate the expression of cLEAP-2 gene in the chicken reproductive organs, as well as in chicken embryos during embryonic development, and to determine whether cLEAP-2 expression in the chicken reproductive organs was constitutive or induced as a response to Salmonella enteritidis infection. RNA was extracted from ovary, oviduct, testis and epididymis of sexually mature healthy and Salmonella infected birds, as well as from chicken embryos until day ten of embryonic development. Expression analysis data revealed that cLEAP-2 was expressed in the chicken ovary, testis and epididymis as well as in embryos during early embryonic development. Quantitative real-time PCR analysis revealed that cLEAP-2 expression was constitutive in the chicken epididymis, but was significantly up regulated in the chicken gonads, following Salmonella infection. In addition, expression of cLEAP-2 during chicken embryogenesis appeared to be developmentally regulated. These data provide evidence to suggest a key role of cLEAP-2 in the protection of the chicken reproductive organs and the developing embryos from Salmonella colonization.     

Comparative efficacy of standard AGID and precipitinogen inhibition test with monoclonal antibodies based competitive ELISA for the serology of <Emphasis Type="Italic">Peste des Petits Ruminants</Emphasis> in sheep and goats

This project was conducted to investigate the comparative efficiency of competitive ELISA (cELISA), standard Agar Gel Immunodiffusion Test (AGID) and Precipitinogen Inhibition Test (PIT) for the diagnosis of Peste des Petits Ruminants (PPR) in Pakistan. To deal with this, serum samples from 198 sheep and 82 goats were collected from three different government livestock farms and all the samples were run simultaneously with the three serological tests. The samples found positive for PPR antibodies through cELISA, AGID and PIT were 96 (34.2%), 60 (21.4%) and 72 (25.7%), respectively. Kappa statistics were applied to evaluate the concordance between the laboratory-based test (cELISA) and field-based tests (AGID and PIT). Kappa statistics scores for cELISA versus AGID and PIT were 0.6343 (95% Confidence Interval CI 0.5231–0.7456) and 0.7134 (95% Confidence Interval CI 0.5987–0.8281), respectively, which indicate a “substantial” agreement between cELISA and AGID and “significant” agreement between cELISA and PIT. AGID and PIT revealed relative diagnostic sensitivities with cELISA of 59.3% and 69.7% and relative diagnostic specificities of 98.3% and 97.2%, respectively. The data suggested that for mass screening and control of PPR, these serological tests proved practical in the absence of cELISA since they have high relative diagnostic specificities and a satisfactory relative diagnostic sensitivities.     

Asexual fungus <Emphasis Type="Italic">Verticillium dahliae</Emphasis> is potentially heterothallic

Verticillium dahliae, a soilborne plant pathogen, causes wilt disease in many important crops. We reported previously that the mating type gene MAT1-2-1 is spread to isolates of this asexual fungus. However, we did not determine whether V. dahliae is homothallic or heterothallic because the opposite mating type gene, MAT1-1-1, had not been identified. In the present study, we identified the MAT1-1-1 gene from an isolate lacking MAT1-2-1 and the mating type idiomorphs of V. dahliae. Each isolate we tested contained either the MAT1-1 or MAT1-2 idiomorph, indicating that the asexual fungus V. dahliae is potentially heterothallic.     

Physio-biochemical parameters: a potential tool for target-selective treatment of haemonchosis in the small ruminants

This study aims to evaluate the conjunctiva colour-based FAMACHA score (FS) coupled with a body condition score (BCS), haemogram and stressor hormone level estimation, in identifying post-mortem (PM)/coproscopically proven individuals wanting therapy for economically important gastrointestinal (GI) helminths, Haemonchus contortus, in the small ruminants. The incidence of haemonchosis was significantly (p < 0.05) higher (60.81%) in the ruminants with FS = 3. The H. contortus count in the animals with FS 2, 3 and 4 was 23.2 ± 0.37, 62 ± 2.5 and 74 ± 3.2 (p < 0.05) [positive correlation (r = 0.841 in goats; r = 0.828 in sheep, p < 0.05)], respectively, with corresponding 2.8 ± 0.15, 2 ± 0.3 and 2 ± 0.16 BCS (negative correlation, p > 0.05). The infected animals of FS 2, 3 and 4 measured 8.2 ± 0.0, 7.5 ± 0.23 and 6.7 ± 0.34 g/dl Hb (r = ?0.452, p = 0.01) in goats/9.3 ± 0.8, 8.6 ± 0.5 and 7.6 ± 0.3 g/dl Hb (r = ?0.511, p = 0.05) in sheep with 21.2, 19.8 ± 1.8 and 17.8 ± 0.2% PCV (r = ?0.369, p = 0.05) in goats/26.7 ± 1.2, 22.2 ± 0.2 and 20.9 ± 0.6% PCV (r = ?0.251, p = 0.03) in sheep, respectively. The FS 2, 3 and 4 infected goats/sheep measured 6.1 ± 0, 7.9 ± 1.0 and 9.5 ± 0.9 (p < 0.05)/5.8 ± 2.3, 6.9 ± 1.2 and 7.8 ± 0.2% (p < 0.05) mid-granulocyte [(r = 0.928 (goats)/0.834 (sheep), p < 0.05], while the cortisol level was 15.6, 23 ± 4.5 and 42 ± 2.3 (p = 0.23)/12.1 ± 0, 15.9 ± 1.2 and 24 ± 3.4 (p = 0.29) μg/dl, respectively. The infected ruminants recorded low (p < 0.05) level of Hb/PCV while high level of mid-granulocytes/cortisol. Specificity of FAMACHA test was maximized (100%) when FS = 4 was considered anaemic, but sensitivity was low (35.29% in goats; 25% in sheep). The false negatives was 5.9 (goat)/12.5 (sheep)% when FS ≥ 3 was considered anaemic. The small ruminants with FS ≥ 3, BCS ≤ 2.5, Hb ≤ 7.5 g/dl (goats)/8.6 g/dl (sheep), PCV ≤ 19.8% (goats)/22.2% (sheep) and mid-granulocyte ≥7.9% (goats)/6.9 ± 1.2% (sheep) can be subjected to target-selective treatment for haemonchosis in the field simultaneously maximizing the economic benefit to the farmers.     

Land contamination by toxic elements in abandoned mine areas in Italy

Purpose

The present paper concerns the distribution and mobility of heavy metals (Cu, Pb, Zn and Fe) in the soils of some abandoned mine sites in Italy and their transfer to wild flora.

Materials and methods

Soils and plants were sampled from mixed sulphide mine dumps in different parts of Italy, and the concentrations of heavy metals were determined.

Results and discussion

The phytoremediation ability of Salix species (Salix eleagnos, Salix purpurea and Salix caprea), Taraxacum officinale and P?lantago major for heavy metals and, in particular, zinc was estimated. The results showed that soils affected by mining activities presented total Zn, Cu, Pb and Fe concentrations above the internationally recommended permissible limits. A highly significant correlation occurred between metal concentrations in soils.

Conclusions

The obtained results confirmed the environmental effects of mine waste; exploring wild flora ability to absorb metals, besides metal exploitation, proved a useful tool for planning possible remediation projects.

     

Determinants of the biodiversity patterns of ammonia-oxidizing archaea community in two contrasting forest stands

Purpose

The variation in soil microbial community patterns is primarily influenced by ecological processes associated with spatial distance and environmental heterogeneities. However, the relative importance of these processes in determining the patterns of soil microbial biodiversity in different successional forests remains unclear.

Materials and methods

Based on the species data from denaturing gradient gel electrophoresis (DGGE) analysis, we described the composition and beta diversity of ammonia-oxidizing archaea (AOA) community, an important functional microbial group in regulating nitrogen cycle, in a middle-succeed stand (60 years of secondary succession) and an undisturbed native stand in a subtropical forest in southern China. The composition pattern was examined using a multi-response permutation procedure (MRPP), and the beta diversity was described using the Sørensen index. The relative influence of edaphic, vegetational, spatial, and topographical factors on AOA composition and beta diversity was assessed by variation partitioning and multiple regression on distance matrices (MRM), respectively.

Results and discussion

We did not find any stand-specific patterns in AOA community composition in the two stands; however, the influential variables were different between the two stands; 7.3 and 4.5 % of the total variation in AOA community composition could be explained by edaphic (i.e., available potassium and total phosphorus) and spatial variables, respectively, in the middle-succeed stand, while 3.7 and 2.8 % of the variation were explained by spatial variable and available phosphorus, respectively, in the native stand. Soil total phosphorus influenced the beta diversity of AOA community most in the middle-succeed stand, while genetic distance of tree species was found to be the most important factor in driving the beta diversity pattern in the native stand.

Conclusions

Soil nutrients influenced the beta diversity of AOA community in the middle-succeed stand more than that in the native stand, while vegetation is more important in the native stand. The substantial unexplained variations were possibly due to the effects of other unmeasured variables. Nevertheless, dispersal process is more important in controlling AOA community composition in the native stand, while processes associated with environmental heterogeneities are more important in the middle-succeed stand in this subtropical forest.

     

Stability of soil organic carbon and potential carbon sequestration at eroding and deposition sites

Purpose

This study aims to explore the dynamics of the factors influencing soil organic carbon (SOC) sequestration and stability at erosion and deposition sites.

Materials and methods

Thermal properties and dissolved aromatic carbon concentration along with Al, Fe concentration and soil specific surface area (SSA) were studied to 1 meter depth at two contrasting sites.

Results and discussion

Fe, Al concentrations and SSA size increased with depth and were negatively correlated with SOC concentration at the erosion site (P?<?0.05), while at the deposition site, these values decreased with increasing depth and were positively correlated with SOC concentration (P?<?0.05). TG mass loss showed that SOC components in the two contrasting sites were similar, but the soils in deposition site contained a larger proportion of labile organic carbon and smaller quantities of stable organic carbon compared to the erosion site. SOC stability increased with soil depth at the erosion site. However, it was slightly variable in the depositional zone. Changes in SUVA₂₅₄ spectroscopy values indicated that aromatic moieties of DOC at the erosion site were more concentrated in the superficial soil layer (0–20 cm), but at the deposition site they changed little with depth and the SUVA₂₅₄ values less than those at the erosion site.

Conclusions

Though large amounts of SOC accumulated in the deposition site, SOC may be vulnerable to severe losses if environmental conditions become more favorable for mineralization in the future due to accretion of more labile carbon. Deep soil layers at the erosion site (>30 cm deep) had a large carbon sink potential.

     

Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Prevalence of antimicrobial residues in raw table eggs from farms and retail outlets in Enugu State,Nigeria

The use of antimicrobial agents in poultry production results in their accumulation in the body tissues and products such as milk and egg. The subsequent accumulation of these drugs and their metabolites in body cells is known as drug residue. This study was designed to determine the prevalence of antimicrobial residues in eggs from poultry farms and retail outlets in Enugu State, Nigeria. Eggs from 25 selected commercial farms and ten retail outlets were screened for the prevalence of antimicrobial residue. Also, structured questionnaires were administered to 25 commercial farms in the state to determine the management practices and the most widely used antimicrobial drugs in farms and possible association between the management practices and the occurrence of antimicrobial residues in eggs from these farms. All the 25 farms surveyed use oxytetracycline. Eggs from nine of the surveyed farms tested positive for antimicrobial residue and three of the ten surveyed farms also tested positive for antimicrobial residue. No association was observed (p 0.05; Fisher’s exact test) between the occurrence of antibiotic residues in eggs and farm size, feed source and housing systems. This study was able to demonstrate the presence of antimicrobial residues in eggs destined for human consumption. Drugs like nitrofurans which has been banned for use in food animals are still very much in use in Enugu State, Nigeria. Antibiotics given as feed additives may give rise to drug residues in food animals.     

Induced morphological mutants in soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill]

The seeds of two soybean cultivars viz., Pusa-16 and PK-1042 were treated with 15, 30, and 45 kR of gamma rays, three concentrations of ethyl methane sulphonate (EMS) (0.1%, 0.2%, and 0.3%), and their combinations. Various morphological mutants were recorded in M₂ and M₃ generations. These morphological mutants were named on the basis of the part of the plant body affected. Eight different morphological mutants, i.e., tall, dwarf, gigas, black pod, smooth pod, black spotted seed, bold seeded, and early maturing were isolated. Among them, early maturing and bold seeded mutants had the potential to be incorporated into breeding programs.