Development of PCR-based tests for the identification of North American isolates of epizootic haemorrhagic disease virus.

A serogroup-specific polymerase chain reaction (PCR) assay and isolate identification strategies (restriction endonuclease analysis (REA) and nucleotide sequencing) were developed for the detection of North American isolates of epizootic haemorrhagic disease virus (EHDV). PCR primers (EHDV-pr4, EHDV-pr5) were designed to hybridize to the L3 gene of a North American isolate of EHDV serotype 1. Total nucleic acid was extracted from preparations of infected tissue culture and PCR was performed using a cDNA-PCR kit, according to the manufacturer's specifications. The PCR assay generated a 459 base pair product from North American isolates of EHDV serotypes 1 and 2, while bluetongue virus (BTV) serotypes 10, 11, 13, and 17, and cell controls, failed to demonstrate PCR products. Slight modifications allowed for the PCR detection of EHDV-1 and -2 in white-tailed deer blood (Odocoileus virginiatus); PCR fragments were not amplified from uninfected deer blood. A number of restriction endonucleases and sequencing primers were evaluated for their utility in isolate identification experiments. Specifically, REA employing HincII and cycle sequencing with an internal primer (EHDV-1-pr3) proved most successful for identifying isolate-specific genome markers. The techniques presented are expected to prove valuable for rapid and specific detection of possible future EHDV incursions in wild and domestic animal species.     

Assessment of biodegradability and oxidation stability of mineral, vegetable and synthetic oil samples

This study proposes to evaluate rapid methodologies to estimate oxidative stability and biodegradability for mineral, vegetable and synthetic oil samples, using quick and simple experiments. The synthetic oil sample was obtained from castor oil, so the vegetable oil chosen for this evaluation was a sample of the crude castor oil source. The oxidative stability tests described enabled the comparison of synthetic lubricant samples and showed that their stability was lower than that of petroleum-based oil. However, the physico-chemical properties, such pour points and viscosity index, were improved and potentially interesting for lubricant applications. The biodegradability experiments were carried out using a model of bio-kinetics. These studies proved that synthetic lubricant samples were easily degradable (similar to crude castor oil) and showed half-life significantly lower than those of the mineral oil samples.     

Epizotiology and phylogeny of equine arteritis virus in hucul horses

The aim of the study was to determine the situation of equine arteritis virus (EAV) infections in hucul horses. A total of 176 horses (154 mares and 22 stallions) from the biggest hucul horse stud in Poland were tested. Antibodies against EAV were detected in 97 (55.1%) horses. The EAV seroprevalence among mares was 53.2% while in stallions - 68.2%. The percentage of positive mares increased with their age, thus amongst the mares of less than 2 years of age the percentage was 32.5%, while in the group of 3-5 years old increased to 59.4% and in the mares in the age of 6-10 years and older than 10 years 89.5% and 95% were seropositive, respectively. Among 11 seropositive stallions five were supposed to be shedders of EAV with their semen. It is likely that those persistently infected stallions were the reservoirs of the virus in the stud. Genetic studies using of ORF5 gene showed high homology between the viruses detected in the semen of those stallions what suggested lateral transmission between the stallions sharing the same stable. Persistent infection in an immature stallion, which has not yet been used for breeding, was established as a result of infection via respiratory route. Phylogenetic analysis confirmed that all hucul viruses shared the same ancestor and as most of EAV strains dominating in Polish horse population belonged to the European origin EAV subgroup (EU-1).     

Changes in biochemical properties of cow manure during processing by earthworms (Eisenia andrei, Bouché) and the effects on seedling growth

The biochemical changes in fresh cow manure caused by the earthworm Eisenia andrei (Bouché) were measured over a period of four months, under controlled laboratory conditions. Earthworms were introduced into each of four plastic containers (0.4 × 0.27 × 0.15 m) containing fresh cow manure (2500 g), and four containers containing manure but without earthworms served as controls. Earthworms reduced the pH and decreased the moisture content in the manure. The C:N ratio of the manure with or without earthworms decreased progressively from 36 to 21. The ash and total nitrogen contents increased greatly for a few weeks after the introduction of earthworms, reflecting a rapid breakdown of carbon compounds and mineralization of nitrogen by the earthworms. CO₂ evolution decreased rapidly (44%) one week after the introduction of earthworms, and continued at a lower rate throughout the 17 weeks (51% reduction as compared to 22% without earthworms), indicating increasing stability of the organic matter. Earthworms reduced microbial biomass early in the process, but enhanced nitrogen mineralization and increased the rates of conversion of ammonium-nitrogen into nitrate. The major general effect of earthworms on the organic wastes was to accelerate the maturation of the organic wastes as demonstrated by enhanced growth of lettuce and tomato seedlings.     

Hepatitis E Virus Antibody Prevalence in Wildlife in Poland

Hepatitis E is an important public health problem mostly in developing but occasionally also in industrialized countries. Domestic and wildlife animals are considered reservoirs of the hepatitis E virus (HEV). Since no information on the prevalence of autochthonous HEV infections in human and animal in Poland is available, the aim of the study was to investigate the HEV seroprevalence of different wildlife species as potential virus reservoirs in the country. No HEV antibodies were found in any of the sera collected from the red deer (Cervus elaphus), European bison (Bison bonasus), roe deer (Capreolus capreolus), elk (Alces alces), fallow deer (Dama dama), sika deer (Cervus nippon), Tatra chamois (Rupicapra rupicapra tatrica) or brown bear (Ursus arctos). HEV‐specific antibodies were detected in 44.4% (95% CI 38.3–50.7) serum samples originated only from wild boars. The percentage of seropositive wild boars differed significantly between the provinces and was positively correlated with the wild boar density and rurality of the area. This study showed that HEV circulates among wild boar population in Poland, and this species should be considered as an important reservoir of the virus.     

Earthworm-Processed Organic Wastes as Components Of Horticultural Potting Media for Growing Marigold and Vegetable Seedlings

We germinated and grew tomato, pepper, lettuce, and marigold seedlings in a standard commercial soilless plant growth medium (Metro-Mix 360), and in coir/perlite and peat/perlite-based container media substituted with 10% or 20%, by volume, of vermicompost derived from pig manure or food wastes. Half of the treatments were watered with liquid inorganic fertilizer while the other half received only water. Germination rates of tomato, pepper, lettuce, and marigold seeds in the coir/perlite mixture did not differ significantly from that in Metro-Mix 360. However, the germination rate of tomato, pepper and lettuce seedlings was very low in the peat/perlite mixture. Substituting some of the peat/perlite mixtures with equal amounts of vermicomposts, particularly pig manure vermicompost, enhanced germination rates greatly, making it comparable to that in the commercial medium (Metro-Mix 360). Pepper, lettuce, and marigold seedlings grown in Metro-Mix 360, which already contains a starter nutrient fertilizer in its formulation, had greater root and shoot dry weights than those grown in the control media (coir/perlite mix and peat/perlite mix). Substituting coir/perlite and peat/perlite mixtures with 10% or 20% of either vermicompost enhanced the growth of seedlings significantly, resulting in an overall plant growth as good as and sometimes better than that in Metro-Mix 360. When the plants were provided daily with a complete fertilizer solution, marigold seedlings in peat-based substrate with 20% pig waste vermicompost, and lettuce seedlings in both coir and peat-based substrates, mixed with 20% food wastes vermicompost, produced greater shoot dry weights than those grown in the commercial potting medium. The growth enhancements tended to be greater in peat/perlite-based mixes than in coir/perlite-based mixes, more so with the addition of pig manure vermicompost than with food waste vermicompost. Earthworm-processed pig manure and food wastes would be suitable materials for inclusion into the formulation of soilless potting media, since substitution of these media with relatively low concentrations of vermicomposts can promote plant growth.     

Application of PCR technique in diagnosis of small ruminant lentivirus infection in sheep and goats

Detection of small ruminant lentiviruses (SRLVs) in sheep and goats usually relies on serological testing. In this study, we evaluated semi-nested PCR and nested PCR techniques applied as a diagnostic tool for detection of maedi-visna virus (MVV) and caprine arthritis-encephalitis virus (CAEV) in naturally infected sheep and goats, respectively. The examination of 193 ovine and 85 caprine serum samples by the ELISA revealed the presence of specific antibodies in 133 (69%) and 18 (21.2%) animals, respectively. Presence of proviral DNA was manifested in 103 (53.4%) sheep and 12 (14.2%) goats. Despite the relatively lower sensitivity of PCR, the fact of detection of proviral DNA in 19 out of 60 ovine samples and 7 out of 67 caprine samples collected from animals previously negative by ELISA was noteworthy. In conclusion, the data demonstrated that combinations of both ELISA and PCR might afford optimal detection of SRLVs infection.     

Effect of Syrian Indigenous Arbuscular Mycorrhizal Fungi in Combination with Manure on the Growth of Cotton (Gossypium hirsutum L.)

A pot experiment was conducted to evaluate the effect of indigenous arbuscular mycorrhizal fungi (AMF) and the synergy of indigenous AMF and sheep manure (SM) on cotton growth and nitrogen and phosphorus uptake. AMF were a mixture of Glomus viscosum, Glomus mosseae, and Glomus intraradices initially isolated from a Syrian cotton field. Dry biomass was enhanced significantly by AMF and was higher at AMF plus SM treatment compared to control. Cotton plants showed a significant dependency to indigenous AMF, which was 52% in the AMF treatment. Plant concentrations of nitrogen (N)and phosphorus (P) were significantly higher in mycorrhizal than nonmycorrhizal plants. Maximum plant N and P uptake was found in the treatment of AMF inoculation with SM, which was significantly higher by 202% and 397% over control, respectively. Indigenous AMF was successful in colonizing cotton roots and when combined with SM resulted in better plant growth and N and P uptake.     

Specific immunoprophylaxis of some viral diseases in cattle

Specific immunoprophylaxis has played a key role in prevention and control of animal infectious diseases for many years. Vaccines are commonly used to diminish economic losses in animal production caused by infectious agents. Currently, respiratory diseases are the main health problem in cattle. BHV1, BRSV and BVD-MD viruses are among the most important pathogens worldwide. They can cause a disease themselves or they can be one of many agents causing a respiratory syndrome. The following article describes the specific immunoprophylaxis of cattle viral diseases caused by such viruses.     

Molecular epizootiology of equine arteritis virus isolates from Poland

Phylogenetic analysis was performed on the sequences of 44 Polish isolates of equine arteritis virus that were isolated from the semen of stallions from national and private studs, collected during 2001--2005. These sequences were also compared with 41 reference strains previously described and commonly used in phylogenesis. On the basis of the nucleotide sequence analysis of the ORF5 gene, encoding the glycoprotein GP5, it was demonstrated that the Polish EAV isolates belonged to two subgroups and showed the closest relationship to the European strains. Similar results were obtained using the nucleotide sequences of the ORF7 gene. The nucleotide identity between the ORF5 and ORF7 sequences of all Polish isolates was in the range of 80.1-99.0% and 93.6-100%, respectively. The analysis of genetic diversity within the ORF5 sequences enabled a retrospective epizootic investigation. This study suggested that some of the EAV shedding stallions were probably infected before they were moved to Poland.