A promising new ELISA diagnostic test for cattle babesiosis based on Babesia bigemina Apical Membrane Antigen-1.

Babesiosis due to Babesia bigemina is a relevant tick‑borne disease, affecting cattle worldwide. Many surface proteins of the pathogen including the Apical Membrane Antigen 1 (AMA‑1) ‑ have been analysed for vaccine and diagnostic purposes. This study focused on B. bigemina AMA‑1 and on its use for the assessment of diagnostic tests. After bioinformatic analyses, AMA‑1 codifying region was amplified and cloned into an expression vector used to induce protein synthesis in Escherichia coli cells. AMA‑1 was purified by affinity chromatography and used to set up the best condition for an ELISA protocol. Bovine field sera positive to B. bigemina were used to evaluate the presence of anti‑AMA‑1 antibodies. In order to verify the assay specificity, sera positive to Babesia bovis or to the piroplasm Theileria annulata were also included. Significant differences were obtained between sera negative to both B. bigemina and B. bovis and samples positive to B. bigemina, to B. bovis or to both pathogens. No significant reaction was observed with T. annulata positive sera. The results showed that AMA‑1 protein is suitable to be used as antigen in diagnostic assays for babesiosis diagnosis in cattle, as it does not show any cross reaction with anti-T. annulata antibodies

User's manual for the Shuttle Electric Power System analysis computer program (SEPS), volume 2 of program documentation

The Shuttle Electric Power System Analysis SEPS computer program which performs detailed load analysis including predicting energy demands and consumables requirements of the shuttle electric power system along with parameteric and special case studies on the shuttle electric power system is described. The functional flow diagram of the SEPS program is presented along with data base requirements and formats, procedure and activity definitions, and mission timeline input formats. Distribution circuit input and fixed data requirements are included. Run procedures and deck setups are described

Noise Properties of Two Mutually Coupled Spin-Transfer Nanooscillators in the Phase Locking Regime

Introduction. Today, many research endeavors are devoted to the miniaturization of microwave sources. One of the promising approaches is the use of magnetic nanostructures (spintronics elements), providing a wide range of frequency tuning and low power consumption. The main disadvantage of spintronics generators (spintransfer nanoscillators ‒ STNO) is a low output power of generated oscillations (tens of nanowatts and less). A possible solution is to sum up the power of many STNOs in a mutual synchronization mode.Aim. The investigation of noise properties of two connected STNOs with identical and non-identical parameters in a phase synchronization mode.Materials and methods. A model was developed of two STNOs interconnected by spin waves taking into account thermal noises. Spectral power densities of the amplitude and phase noise were obtained by the method of effective linearization.Results. Dependencies were obtained in a general form for attenuation coefficients of the amplitude and phase fluctuations of noise sources for each STNO. Three cases of synchronization were considered: completely identical STNOs, two identical STNOs but with different oscillation frequencies, and two non-identical STNOs, differing in an allowance of self-excitation by frequencies and amplitudes of the oscillations. It was possible to obtain a gain in the amplitude and phase noise for two identical STNOs. In this case, an increase in the allowance of self-excitation led to a decrease in the level of phase and amplitude noise.Conclusion. This analysis of the attenuation coefficients for non-identical STNOs demonstrates the possibility of improving the noise properties of each of the generators. In this case, the best noise value is obtained for an STNO with greater stability in a stand-alone mode.Introduction. Today, many research endeavors are devoted to the miniaturization of microwave sources. One of the promising approaches is the use of magnetic nanostructures (spintronics elements), providing a wide range of frequency tuning and low power consumption. The main disadvantage of spintronics generators (spintransfer nanoscillators ‒ STNO) is a low output power of generated oscillations (tens of nanowatts and less). A possible solution is to sum up the power of many STNOs in a mutual synchronization mode.Aim. The investigation of noise properties of two connected STNOs with identical and non-identical parameters in a phase synchronization mode.Materials and methods. A model was developed of two STNOs interconnected by spin waves taking into account thermal noises. Spectral power densities of the amplitude and phase noise were obtained by the method of effective linearization.Results. Dependencies were obtained in a general form for attenuation coefficients of the amplitude and phase fluctuations of noise sources for each STNO. Three cases of synchronization were considered: completely identical STNOs, two identical STNOs but with different oscillation frequencies, and two non-identical STNOs, differing in an allowance of self-excitation by frequencies and amplitudes of the oscillations. It was possible to obtain a gain in the amplitude and phase noise for two identical STNOs. In this case, an increase in the allowance of self-excitation led to a decrease in the level of phase and amplitude noise.Conclusion. This analysis of the attenuation coefficients for non-identical STNOs demonstrates the possibility of improving the noise properties of each of the generators. In this case, the best noise value is obtained for an STNO with greater stability in a stand-alone mode

Ovine Catarrhal fever (bluetongue): Analysis of Culicoides species in seropositive farms

Bluetongue (BT) is an orbiviral disease of wild and domestic ruminants, mainly sheep. In Sicily, the first Bluetongue outbreak occurred in October 2000; there have been 76 recorded outbreaks so far. The National Surveillance Plan, based on European Union Commission Decision 138/2001/CE, establishes serological and entomological surveys. This plan consists of controls of seronegative cattle, called 'sentry' as indicators for the presence and circulation of virus in defined areas. To check the seroconversions, the regional territory has been subdivided in 400 km2 areas including 58 seronegative cattle, periodically checked by serological tests. All positive sera have been tested to detect the specific serotype by the National Reference Centre for Exotic Diseases (CESME) at the Istituto Zooprofilattico Sperimentale Abruzzo e Molise in Teramo (IZS Teramo). Moreover, entomological surveillance has been implemented in seropositive herds, to investigate the presence of insect vectors belonging to Culicoides genus. The goal of the present communication is to report on the different species of Culicoides found in the farms with Bluetongue virus and to investigate on the probable role of new competent vectors. This paper concerns data analysis of 581 light-trap catches collected in 321 farms from 2003 to 2008. We observed that 82% of checked farms were positive for Culicoides spp., and only 10% of the farms were positive for Culicoides imicola. © 2010 Blackwell Verlag GmbH

Reversible switch from hemoglobin A to C in sheep and recovery from anemia following experimental infection with Anaplasma ovis

Anemia causes a change in the type of circulating hemoglobin (Hb) in sheep carrying the βA-globin haplotype, where the Hb A is replaced with Hb C, unlike Hb B. The effect of the substitution of Hb A with Hb C on the recovery from anemia was investigated by comparing the hematological picture of sheep, following experimental infection with Anaplasma ovis. The blood values were obtained from 3 AB and 3 BB Hb sheep after the development of the disease where anemia is a pathognomonic symptom. The expression of the silent gene encoding for Hb C was detected by isoelectric focusing and quantified by high performance liquid chromatography. Both Hb AB genotype and Hb C occurrence were involved in the lower recovery from anemia in the trial

Hemoprotozoa of domestic animals in France: Prevalence and molecular characterization

Very limited information is available on epizootiology of haematozoan infections in French domestic animals. In an attempt to address this issue, prevalence of piroplasmida was studied in carnivores and ruminants, whereas prevalence of Hepatozoon spp. was only investigated in carnivores. In total, 383 animals were included in the survey (namely 116 cats, 108 dogs, 91 sheep and 68 cows). Parasite diagnosis was carried out using molecular methods such as PCR and sequencing of the 18S rRNA gene. In addition, ruminant samples were analyzed with the reverse line blotting technique (RLB). Results of RLB and PCR plus sequencing were in total agreement. In carnivores, haematozoan prevalence was close to 1%. Two cats were infected by H. canis (1.7% prevalence) and one of them was co-infected by Cytauxzoon sp. (0.8%). This represents the first finding of both pathogens in French cats. One dog was infected by H. canis (0.9%) and another by Babesia canis vogeli (0.9%). In ruminants, haematozoan prevalence (piroplasmida) was significantly higher than in carnivores (4.8% in sheep and 8.8% in cow). Theileria ovis was found in 1 sheep, Theileria sp. in 2 sheep, Theileria buffeli in 5 cows and B. major in 1 cow. Evidence presented in this contribution indicates that haematic protozoa are not widely distributed in domestic mammal populations of France

Program manual for the Shuttle Electric Power System analysis computer program (SEPS), volume 1 of program documentation

The Shuttle Electric Power System (SEPS) computer program is considered in terms of the program manual, programmer guide, and program utilization. The main objective is to provide the information necessary to interpret and use the routines comprising the SEPS program. Subroutine descriptions including the name, purpose, method, variable definitions, and logic flow are presented

Serological survey and molecular characterization of theileria annulata in sicilian cattle

Tropical theileriosis is a tick-borne disease caused by hemoprotozoan parasites with considerable veterinary and economic impact worldwide. Ticks transmitting the disease belong to the Haemaphysalis, Rhipicephalus, and Hyalomma genera. The Hyalomma genus is very common in Sicily (Italy) and represents the main Theileria annulata vector in the island. Data concerning the molecular epidemiology of this pathogen are missing in the region. In 2018–2019, blood and serum samples were collected from 480 cows in seven Sicilian farms from four different provinces. Seroprevalence in the farms ranged from 22% to 71%. Three farms were selected for molecular analysis consisting of real-time PCR targeting the almost complete 18S ribosomal RNA (rRNA). Four amplicons per farm were sequenced and phylogenetic analyses were carried out. The four sequences were identical within each farm and showed 92–99% identity with the other farms and with sequences from Genbank. According to the phylogenetic analysis, these three sequences and an additional one from a laboratory-cultured Theileria annulata strain obtained in 1999 belonged to a single T. annulata clade with good bootstrap support with other sequences from Italy, India, and Iran, indicating limited geographical and temporal genetic variability of the parasite. This study represents the first phylogenetic analysis of T. annulata in Sicily, which will be useful to improve the strategies for theileriosis control and prevention

Detection of natural killer T cells in mice infected with Rickettsia conorii

Little information is available regarding the role of natural killer T (NKT) cells during the early stage of Rickettsia conorii infection. Herein, C3H/HeN mice were infected with the Malish 7 strain of R. conorii. Splenocytes from these mice were analysed in the early stage of the infection by flow cytometry and compared with uninfected controls. Our results showed an increase in NKT cells in infected mice. Additionally, NKT interleukin (IL)-17(+) cells increased three days after infection, together with a concurrent decrease in the relative amount of NKT interferon (IFN)-\u3b3(+) cells. We also confirmed a higher amount of NK IFN-\u3b3(+) cells in infected mice. Taken together, our data showed that NKT cells producing Il-17 increased during the early stage of rickettsial infection. These results suggest a connection between IL-17(+) NKT cells and vasculitis, which is the main clinical symptom of rickettsiosi

Acute febrile illness is associated with Rickettsia spp infection in dogs

BACKGROUND: Rickettsia conorii is transmitted by Rhipicephalus sanguineus ticks and causes Mediterranean Spotted Fever (MSF) in humans. Although dogs are considered the natural host of the vector, the clinical and epidemiological significance of R. conorii infection in dogs remains unclear. The aim of this prospective study was to investigate whether Rickettsia infection causes febrile illness in dogs living in areas endemic for human MSF. METHODS: Dogs from southern Italy with acute fever (n = 99) were compared with case–control dogs with normal body temperatures (n = 72). Serology and real-time PCR were performed for Rickettsia spp., Ehrlichia canis, Anaplasma phagocytophilum/A. platys and Leishmania infantum. Conventional PCR was performed for Babesia spp. and Hepatozoon spp. Acute and convalescent antibodies to R. conorii, E. canis and A. phagocytophilum were determined. RESULTS: The seroprevalence rates at first visit for R. conorii, E. canis, A. phagocytophilum and L. infantum were 44.8%, 48.5%, 37.8% and 17.6%, respectively. The seroconversion rates for R. conorii, E. canis and A. phagocytophilum were 20.7%, 14.3% and 8.8%, respectively. The molecular positive rates at first visit for Rickettsia spp., E. canis, A. phagocytophilum, A. platys, L. infantum, Babesia spp. and Hepatozoon spp. were 1.8%, 4.1%, 0%, 2.3%, 11.1%, 2.3% and 0.6%, respectively. Positive PCR for E. canis (7%), Rickettsia spp. (3%), Babesia spp. (4.0%) and Hepatozoon spp. (1.0%) were found only in febrile dogs. The DNA sequences obtained from Rickettsia and Babesia PCRs positive samples were 100% identical to the R. conorii and Babesia vogeli sequences in GenBank®, respectively. Febrile illness was statistically associated with acute and convalescent positive R. conorii antibodies, seroconversion to R. conorii, E. canis positive PCR, and positivity to any tick pathogen PCRs. Fourteen febrile dogs (31.8%) were diagnosed with Rickettsia spp. infection based on seroconversion and/or PCR while only six afebrile dogs (12.5%) seroconverted (P = 0.0248). The most common clinical findings of dogs with Rickettsia infection diagnosed by seroconversion and/or PCR were fever, myalgia, lameness, elevation of C-reactive protein, thrombocytopenia and hypoalbuminemia. CONCLUSIONS: This study demonstrates acute febrile illness associated with Rickettsia infection in dogs living in endemic areas of human MSF based on seroconversion alone or in combination with PCR