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.

     

Soil nitrogen mineralization not affected by grass species traits

Species N use traits was evaluated as a mechanism whereby Bromus inermis (Bromus), an established invasive, might alter soil N supply in a Northern mixed-grass prairie. We compared soils under stands of Bromus with those from three representative native grasses of different litter C/N: Andropogon gerardii (Andropogon), Nassella viridula (Nassella) and Pascopyrum smithii (Pascopyrum); in ascending order of litter quality. Net mineralization (per g soil N) measured in year-long laboratory incubations showed no differences in comparisons of Bromus with two of the three native grasses: Andropogon and Nassella. Higher mineralization in Pascopyrum stands relative to Bromus was consistent with its higher litter quality. However, an unusually high occurrence of an N-fixing legume in Pascopyrum stands, potentially favoring high mineralization rates, confounded any conclusions regarding the effects of plant N use on N mineralization. Instead of an initial flush of net mineralization, as would be expected in laboratory incubation, we observed an initial lag phase. This lag in net N mineralization coincided with high microbial activity (respiration) that suggests strong N limitation of the microbial biomass. Further support for the importance of immobilization initially came from modeling mineralization dynamics, which was explained better when we accounted for microbial growth in our model. The absence of strong differences in net mineralization beneath these grasses suggests that differences in plant N use alone were unlikely to influence soil N mineralization through substrate quality, particularly under strong N control of the microbial biomass.     

Docosahexaenoic Acid Is Naturally Concentrated at the sn-2 Position in Triacylglycerols of the Australian Thraustochytrid Aurantiochytrium sp. Strain TC 20

The Labyrinthulomycetes or Labyrinthulea are a class of protists that produce a network of filaments that enable the cells to glide along and absorb nutrients. One of the main two Labyrinthulea groups is the thraustochytrids, which are becoming an increasingly recognised and commercially used alternate source of long-chain (LC, ≥C₂₀) omega-3 containing oils. This study demonstrates, to our knowledge for the first time, the regiospecificity of the triacylglycerol (TAG) fraction derived from Australian thraustochytrid Aurantiochytrium sp. strain TC 20 obtained using ¹³C nuclear magnetic resonance spectroscopy (¹³C NMR) analysis. The DHA present in the TC 20 TAG fraction was determined to be concentrated in the sn-2 position, with TAG (16:0/22:6/16:0) identified as the main species present. The sn-2 preference is similar to that found in salmon and tuna oil, and differs to seal oil containing largely sn-1,3 LC-PUFA. A higher concentration of sn-2 DHA occurred in the thraustochytrid TC 20 oil compared to that of tuna oil.     

Enhanced molecular identification of Botrytis spp. from New Zealand onions

Botrytis spp. associated with neck rot disease were isolated from New Zealand onions. The fungi were identified using molecular sequences of the ribosomal internal transcribed spacer (ITS) and intergenic spacer (IGS) regions, and the glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene. Analyses of the sequences showed that the majority of the isolates gathered in 2005–07 were B. aclada. A new high resolution melting analysis (HRMA) assay was developed that allowed fast and simple discrimination between B. aclada and other Botrytis spp. causing onion neck rot in New Zealand. To further verify these results, Botrytis isolates from New Zealand onions, stored in the International Collection of Microorganisms from Plants (ICMP), were also examined. Only a single isolate from the ICMP collection was B. aclada while two isolates were B. byssoidea, one B. squamosa and another closely related to Botryotinia porri. Identification of the remaining Botrytis isolates was more difficult; while IGS and ITS sequences indicated a close relationship to B. allii or B. byssoidea, a previously unreported intron insertion was observed at the 3′ end of the ribosomal small subunit gene in these isolates. No evidence of heterogeneity was observed in the G3PDH gene sequences, as might have been expected of the allodiploid B. allii, but the G3PDH sequence ruled out B. byssoidea as the identity of these isolates.     

A study on the synergetic effect of Bacillus amyloliquefaciens and dipotassium phosphate on Alternaria solani causing early blight disease of tomato

European Journal of Plant Pathology - A study of the effect of bioagents and dipotassium phosphate (DPP) and their combination on early blight disease reduction under greenhouse conditions was...     

Rhizobium inoculation of Calopogonium caeruleum

The shade-tolerant cover legume Calopogonium caeruleum is promiscuous in its nodulating habits. In sand culture, symbiotic effectiveness of the strains tested was variable; 6 strains of rhizobia markedly improved shoot yields and 20 increased shoot N content. In pot experiments using cultivated and non-cultivated soils, inoculation gave no significant increase in shoot yields. When grown under rubber in plantation conditions at four localities, shoot dry matter yields, N content and nodulation also were not different from uninoculated plants when sampled for up to 2 yr after planting. This occurred despite the low numbers (< 10 g^?1 soil) of native rhizobia at some sites and an appreciable establishment (> 70% recovery in nodules) by the inoculant strains.     

Rhizobia in tropical legumes—IX. pot and field trials with inoculants for Psophocarpus tetragonolobus (L.) DC.

Antigenically identifiable inoculants for Psophocarpus tetragonolobus were evaluated in three non-sterile soils contained in pots (sandy-clay, Renggam series; a loamy-sand, Sungei Buloh series; silty-clay, Munchong series). Most-probable-numbers of indigenous rhizobia ranged from 4 (Renggam series) to 13 (Munchong series) g^?1. Only two (RRIM 56 and 968) of the eight rhizobia tested formed > 50% of the nodules in all soils. Recovery of two strains (RRIM 968 and UMKL 12) was significantly poorer from the Munchong series soil which had the most indigenous rhizobia and the highest silt plus clay content. In a field trial using a Sungei Buloh series soil containing 700 rhizobia g^?1 capable of nodulating P. tetragonolobus, none of the applied strains formed > 18% of the nodules; two formed no nodules. There were no significant increases in plant yield in response to inoculation in the field trial and in two soils in the pot trials. In Sungei Buloh series soil, RRIM 56 formed 90% of the nodules when the indigenous rhizobia were 5 cells g^?, and 14% when the population was 700 g^?1. This raises the question of the need to inoculate seed sown into soils with high indigenous rhizobial populations, but there was some indication of increasing representation of inoculant strains in nodules with time.     

Acute kidney injury mediated by oxidative stress in Egyptian horses with exertional rhabdomyolysis

The present study was carried out to evaluate the role of oxidative stress in the pathophysiologic process of acute renal failure associated with exertional rhabdomyolysis (ER) in Egyptian horses. ER was tentatively diagnosed in 31 Baladi horses based on case history, physical examination findings and confirmed by elevation of plasma creatine kinase (CK) and urine myoglobin concentrations. According to severity of the condition, the diseased horses were categorized into two main groups; the first group included 18 horses with minimal clinical signs and plasma CK <60 000 IU/L; whereas, the second group included 13 horses with overt clinical signs and plasma CK >100 000 IU/L). It was found that plasma creatol (CTL) was positively correlated (p < 0.01) with plasma malondialdehyde (MDA) (r = 0.775), nitric oxide (NO) (r = 0.768), methyguanididne (MG) (r = 0.995), CK (r = 0.768), urine glucose (r = 0.778), urine protein (r = 0.767), renal failure index (RFI) (r = 0.814) and urine sodium (r = 0.799) and negatively correlated (p < 0.01) with total antioxidant capacity (TAC) (r = −0.795), superoxide dismutase (SOD) (r = −0.815), glutathione peroxidase (GSH-Px) (r = −0.675), Vitamin C (r = −0.830), urine creatinine (r = −0.800), urine/plasma creatinine ratio (r = −0.827) and urine/plasma urea ratio (r = −0.807). The correlation between these biochemical variables might suggest a possible role of oxidative stress in renal injury associated with severe rhabdomyolysis in horses. It is suggested that exaggeration of oxidative stress associated with increased muscle membrane leakage plays a key role in acute kidney injury in Baladi horses with severe rhabdomyolysis.