Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Sulfur in soils

Sulfur (S) deficiency of crops, which has been reported with increasing frequency over the past two decades on a worldwide scale, is a factor that reduces yield and affects the quality of harvested products. Especially in Western European countries, incidence of S deficiency has increasingly been reported in Brassicaceae. For this reason, more attention should be paid to the optimization of S‐fertilizer application, in order to cover plant S requirements whilst minimizing environmental impacts. In soils, S exists in inorganic and organic forms. While sulfate (SO ), which is a direct S source for plants, contributes up to 5% of total soil S, generally more than 95% of soil S are organically bound. Organic S is divided into sulfate ester and carbon‐bonded S. Although not directly plant‐available, organically bound S may potentially contribute to the S supply of plants, especially in deficiency situations. Sulfur turnover involves both biochemical and biological mineralization. Biochemical mineralization, which is the release of SO from the ester sulfate pool through enzymatic hydrolysis, is controlled by S supply, while the biological mineralization is driven by the microbial need for organic C to provide energy.     

Energetics of honeybee swarm thermoregulation

Honeybee swarms regulate their core temperature at a high set point (near 36 degrees C) and the mantle at a low set point (near 15 degrees C). The temperature gradient from core to mantle permits considerable energy economy, and it is abolished only shortly before swarm takeoff.     

cDNA-cloning and expression of VP1-specific sequences of foot-and-mouth disease virus types A5 and O1

The RNA genome of foot- and mouth disease virus strains A5 Westerwald and O1 Lausanne has been reverse-transcribed and cloned in lambdaphages or plasmids. Identification of cDNA-clones containing VP1-specific sequences was achieved by hybridization, restriction mapping, and sequence analysis. VP1-coding cDNA-fragments were subcloned into the expression vector pEX which led to synthesis of fusion proteins with beta-galactosidase. These fusion proteins reacted with anti-VP1 antibodies on a Western blot, but were not capable of inducing neutralizing antibodies to mice. This seemed to suggest a tertiary structure of the VP1-epitopes unlike those of native VP1. Other attempts are discussed to construct VP1-fusion proteins folding similarly to the native viral protein structure.     

Temporal dynamics of nutrient and carbohydrate distribution during crop cycles of Rosa spp. ‘Kardinal’ in response to light availability

Rose plants that are flush harvested exhibit episodic growth patterns. During these crop cycles little biomass accumulation occurs immediately following harvest; and as new shoots emerge a period of rapid shoot growth and biomass accumulation occurs. The temporal changes in whole-plant nutrient and carbohydrate distribution during these crop cycles and the role of storage in new shoot growth are not well documented. The objective of this project was to quantify N, P, K, and total non-structural carbohydrates (TNC) distribution in roots, base stems, base leaves, and new shoots during crop cycles in response to light availability. Plants were grown in solution culture under high or low light (mean daily light integral 45.3 or 13.1 mol m⁻² d⁻¹, respectively) during 30–35 day crop cycles. Every five days destructive sampling was used to determine biomass and N, P, K, and TNC concentration of rose plant compartments. N and TNC accumulated in base plant compartments during the first ten days of the crop cycles. N, P, K, and TNC in base plant compartments declined during days 10–25 during a crop cycle concurrent with the rapid growth of flower shoots. N, P, and K storage in base plant parts represents 27, 22, and 24% of the potential N, P, and K required by flower shoots under high light; and 19, 21, and 22% of requirements under low light. TNC storage in base plant parts represents 4–10% of the final biomass of flower shoots. Mobilization of N, P, K, and TNC stored from base plant parts appears to be important during the stage of rapid flower shoot growth when absorption by roots or photosynthesis by shoots was insufficient to meet flower shoot demands. Plant carbohydrate status was improved under high light conditions; storage of N and TNC declined under low light.     

An investigation into the bacterial contamination of goniolenses in use in clinical practice

Objectives To report the incidence and evaluate the clinical significance of goniolens bacterial contamination in clinical use in dogs with three different usage protocols and one with an added cleaning protocol. Animals Studied and Methods Three groups of twenty dogs undergoing gonioscopy at a private practice in the UK had the goniolenses swabbed for bacteriology culture and identification prior to placement on the cornea. Three protocols of lens use, with 2 different types of goniolens, were studied. One protocol was then repeated with 21 dogs with a lens cleaning protocol prior to storage. Results Low levels of bacterial contamination were found in all 3 initial groups (10‐15%). No correlation was found between usage protocol used and rate of contamination and no correlation was found between length of storage between use and contamination. All bacteria cultured were considered naturally occurring commensals for the canine eye and environment. The group with a cleaning protocol had a 4.7% contamination rate. This was not statistically different from the non‐cleaning groups. Conclusions The rate of bacterial contamination of goniolenses in clinical practice is low and the bacterial contaminants consist of commensal bacteria, unlikely to be of detriment to the eye. Minimal contamination of the goniolenses was found and this did not appear to be of clinical significance. The introduction of a simple cleaning protocol did not produce a statistically significant reduction in bacterial contamination.     

Carnivorous fungi from Cretaceous amber

Carnivorous fungi dating back to the age of the dinosaurs have been found fossilized in circa-100-million-year-old amber. The fossil fungi used hyphal rings as trapping devices and are preserved together with their prey, small nematodes. The excellent preservation in amber allowed comparison with extant groups: On the basis of the mode of ring formation and the dimorphic mode of life, the fossils cannot be assigned to any recent carnivorous fungus, providing evidence that different groups occupied this ecological niche in the Cretaceous and that trapping devices were developed independently multiple times in the course of Earth history.     

Cesium-137 Monitoring Using Mosses from Macedonia,N. Greece

137Cs activities in mosses collected from four sites in Macedonia, N. Greece were measured for regular periods of time during the last decade following the reactor accident in Chernobyl. Samples taken in 1985 allow to determine the 137Cs contamination before the accident. A clear increase of the cesium interception was observed for about two years after the accident. Later a gradual decrease was detected which depended on various factors such as moss species, location, growth rate and substrate. Higher 137Cs amounts were observed in epilithic (Grimmia) in comparison to epiphytic or epigeic ones ( Encalypta, Tortula). From the studied biotopes Mt. Vermion presented the highest 137Cs activities. The high 137Cs values found 10 yr after Chernobyl suggest that mosses are effective and inexpensive biological detectors of the distribution and burden of fallout radionuclides.     

Variability in plasma membrane integrity, mitochondrial membrane potential and acrosomal status before and after cryopreservation of bull sperm

In the present study variability of bull sperm concerning percentages of sperm with intact plasma membranes (PMI), high mitochondrial membrane potential (HMMP) and positive acrosomal status (PAS) before and after cryopreservation (vKK; nKK) between bulls and between ejaculates within bulls was examined. Studies were performed on 4 semen samples each of 20 Deutsche Fleckvieh bulls. VKK-Values were 76.5% +/- 9.6% (PMI) 68.3% +/- 8.9% (HMMP) and 9.8 +/- 5.1% (PAS) and nKK-values were 38.1 +/- 14.0% (PMI), 38.2 +/- 14.0% (HMMP) and 30.9 +/- 12.1% (PAS). After freezing, variabilities in sperm parameter values between bulls (nKK: PMI: 49.8%, HMMP 52.1% and PAS: 56.6%) were nearly quite as high or higher than variabilities between ejaculates (nKK: 50.2%, 47.9% and 43.4%). VKK-values of PMI, HMMP and PAS were only fairly to moderately related (0.36 < r < 0.53; P < 0.05) to nKK-values. The results show that PMI, HMMP and PAS did not only vary between bulls, but also between ejaculates within bulls. As there are no high relationships in these sperm parameters between times before and after cryopreservation, each ejaculate should be examined after cryopreservation in order to receive a reliable information about the quality of cryopreserved sperm.