Sensibilidade e especificidade do "California Mastitis Test" como recurso diagnóstico da mastite subclínica em relação à contagem de células somáticas

O "California Mastitis Test" (CMT) estima o conteúdo de células somáticas no leite e é interpretado subjetivamente, estabelecendo-se escores que, na maioria dos casos, variam de 1 a 5. O escore 1 indica uma reação completamente negativa e os de 2-5 indicam graus crescentes de resposta inflamatória do úbere, sendo normalmente considerados como indicativos de mastite subclínica. Dependendo da interpretação dos escores, o CMT pode produzir resultados falso-positivos ou falso-negativos. Esse trabalho teve o objetivo de avaliar a sensibilidade e a especificidade do CMT em relação à contagem de células somáticas (CCS). Foram utilizadas 3.012 amostras de leite provenientes de 760 vacas Holandesas ou mestiças Holandês-Zebu, pertencentes a 15 rebanhos. Todas as amostras foram submetidas ao CMT e processadas para CCS em equipamento Fossomatic 90. Os valores médios de CCS (x 1.000 células/ml) obtidos para os escores de CMT foram 1 (79,9), 2 (333,5), 3 (670,3), 4 (1.354,0) e 5 (4.455,6). Três opções de interpretação (doente/não-doente) para o CMT foram testadas, em relação aos valores de CCS, iniciando com 100.000 células/ml: (a) 1 versus 2, 3, 4, e 5; (b) 1 e 2 versus 3, 4 e 5; (c) 1, 2, 3 versus 4 e 5. As sensibilidades do CMT em identificar corretamente quartos mamários acima de 200.000 células/ml foram 79%, 61% e 34%, para as opções a, b e c, respectivamente. Para identificar corretamente contagens acima de 500.000 células/ml, as sensibilidades do CMT, para as opções a, b e c, foram, respectivamente: 93%, 82% e 54%. A sensibilidade do CMT em identificar quartos mamários com mastite subclínica foi adequada (acima de 80%) quando a interpretação do teste foi mais rigorosa (opções a e b). A interpretação da reação 3 como negativa (opção c) só alcançou sensibilidade de 80% para contagens entre 1.200.000 e 1.400.000 células/ml. As especificidades do CMT, para CCS de 200.000 e 500.000 foram, respectivamente, 90% e 80% (opção a), 97% e 90% (opção b) e 99% e 97% (opção c).

A conglutination test for rapid detection of antibodies against Babesia bigemina

A rapid conglutination test (RCT) with performance comparable to the indirect fluorescent antibody technique (IFAT) was developed to detect antibodies against Babesia bigemina (B. bigemina-RCT). The B. bigemina-RCT is a sensitive, specific, economical, and rapidly performed serological test suitable for field application or minimally equipped laboratories. This test had a sensitivity of 90.9%, and specificity of 97.6%, compared to IFAT, which showed for the same parameters respectively, 98.3% and 99.7%. The early detection of anti- B. bigemina immunoglobulins by RCT in experimental infections was nearly parallel to that of IFAT. Cross reactions were observed with sera from calves experimentally infected with Babesia bovis (1.8%) and with Anaplasma marginale (1.2%). RCT antigen prepared with non parasitized erythrocytes (negative antigen) showed 1.5%, 3.5% and 2.2% of positive reactions with sera from animals experimentally infected with B. bigemina, B. bovis and A. marginale. However, none of the sera from animals of endemic areas for babesia infection resulted in positive reactions with the negative antigen. Considering these results and shelf life over six months, the B. bigemina-RCT could be used for epidemiological surveys and evaluation of control measures against this species of Babesia.

Multibody Simulation Model of the Roller Test Rig

Multibody simulation model of the roller test rig is presented in this work. The roller test rig consists of a paper machine’s tube roll supported with a hard bearing type balancing machine. The simulation model includes non-idealities that are measured from the physical structure. These non-idealities are the shell thickness variation of the roll and roundness errors of the shafts of the roll. These kinds of non-idealities are harmful since they can cause subharmonic resonances of the rotor system. In this case, the natural vibration mode of the rotor is excited when the rotation speed is a fraction of the natural frequency of the system. With the simulation model, the half critical resonance is studied in detail and a sensitivity analysis is performed by simulating several analyses with slightly different input parameters. The model is verified by comparing the simulation results with those obtained by measuring the real structure. Comparison shows that good accuracy is achieved, since equivalent responses are achieved within the error limit of the input parameters.

Evaluation of the indirect fluorescent antibody test and modified agglutination test for detection of antibodies against Toxoplasma gondii in experimentally infected pigs

The study determined the sensitivity and specificity of the indirect fluorescent antibody test (IFAT) and modified agglutination test (MAT) for anti-Toxoplasma gondii antibody detection by analyzing sera from 46 experimentally infected pigs. Values for sensitivity were 95.7% (confidence interval 95%: 84.0-99.2%) and for specificity 97.8% (confidence interval 95%: 87.0-99.9%) in both tests. There was an optimum agreement of results between IFAT and MAT evidenced by a Kappa test of 0.86. These results validate these tests for the detection of T. gondii infection in pigs. IFAT and MAT despite methodologies with different characteristics and readings have similar accuracy in pig serum samples.

Evaluation of tear production with modified Schirmer tear test-1 during the neonatal period in cats

The study aimed to evaluate tear production by means of modified Schirmer tear tes-1 (mSTT-1) in neonate cats. Likewise, correlation between mSTT-1 and STT-1 was assessed in vitro. Standard SST strips were cut in half and after eye lid opening, tear production of neonates (n=15) was daily measured in both eyes (mSTT-1), until the 7th day, and at day 14, 21, and 28. Animals were daily weighted until 28 days of age. Results were compared statistically (P<0.05). During the first 7 days, the overall mSTT-1 mean was 0.76 wetting/minute. Significant differences between right and left eyes were not observed at any time point (P=1.00). Tear secretion increased significantly, from the 14th to 28th day, in comparison with 7 first days (P<0.05). Positive correlation between maturity parameters and tear secretion was observed (P<0.0001). Distance between slopes of each strip changed significantly (P<0.0001). It was concluded that tear secretion in the neonatal period of cats is very below the reference values described for young and adults of the same species. It is not possible to extrapolate results obtained with mSTT-1 to standard STT-1.

Low dose of dichloroacetate infusion reduces blood lactate after submaximal exercise in horses

The acute administration of an indirect activator of the enzyme pyruvate dehydrogenase (PDH) in human athletes causes a reduction in blood lactate level during and after exercise. A single IV dose (2.5m.kg-1) of dichloroacetate (DCA) was administered before a submaximal incremental exercise test (IET) with five velocity steps, from 5.0 m.s-1 for 1 min to 6.0, 6.5, 7.0 and 7.5m.s-1 every 30s in four untrained mares. The blood collections were done in the period after exercise, at times 1, 3, 5, 10, 15 and 20 min. Blood lactate and glucose (mM) were determined electro-enzymatically utilizing a YSI 2300 automated analyzer. There was a 15.3% decrease in mean total blood lactate determined from the values obtained at all assessment times in both trials after the exercise. There was a decrease in blood lactate 1, 3, 5, 10, 15 and 20 min after exercise for the mares that received prior DCA treatment, with respective mean values of 6.31±0.90 vs 5.81±0.50, 6.45±1.19 vs 5.58±1.06, 6.07±1.56 vs 5.26±1.12, 4.88±1.61 vs 3.95±1.00, 3.66±1.41 vs 2.86±0.75 and 2.75±0.51 vs 2.04±0.30. There was no difference in glucose concentrations. By means of linear regression analysis, V140, V160, V180 and V200 were determined (velocity at which the rate heart is 140, 160, 180, and 200 beats/minute, respectively). The velocities related to heart rate did not differ, indicating that there was no ergogenic effect, but prior administration of a relatively low dose of DCA in mares reduced lactatemia after an IET.

Introduction of the T-10 static line parachuting capability to the NH90 helicopter

The purpose of this study was to investigate the suitability of the Finnish Defence Forces’ NH90 helicopter for parachuting operations with the T-10 static line parachute system. The work was based on the Army Command’s need to compensate for the reduction in the outsourced flight hours for the military static line parachuting training. The aim of the research was to find out the procedures and limitations with which the NH90 IOC+ or FOC version helicopter could be used for static line parachutist training with the T-10B/MC1-1C parachutes. The research area was highly complicated and non-linear. Thus analytical methods could not be applied with sufficient confidence, even with present-day computing power. Therefore an empirical research method was selected, concentrating on flight testing supported with literature study and some calculated estimations. During three flights and 4.5 flight hours in Utti, Finland on 1720 September 2012, a total of 44 parachute drops were made. These consisted of 16 dummy drops and 28 paratrooper jumps. The test results showed that when equipped with the floor mounted PASI-1 anchor line, the deflector bar of the NHIndustries’ Parachuting Kit and Patria’s floor protection panels the Finnish NH90 variant could be safely used for T-10B/MC1-1C static line parachuting operations from the right cabin door at airspeed range of 5080 KIAS (90–150 km/h). The ceiling mounted anchor lines of the NHI’s Parachuting Kit were not usable with the T-10 system. This was due to the static lines’ unsafe behaviour in slipstream when connected to the cabin ceiling level. In conclusion, the NH90 helicopter can be used to meet the Army Command’s requirement for an additional platform for T-10 static line parachutist training. Material dropping, the effect of additional equipment and jumping from the rear ramp should be further studied.

Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test

Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM) and moxidectin (MOX) effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT). Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL). The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50) dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL), identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001) and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA) guidance.

Development and application of polymerase chain reaction test for detection of Conidiobolus lamprauges

Conidiobolomycosis is a granulomatous disease caused by the fungus Conidiobolus spp. in humans and animals. Traditional technique for diagnosis of the disease is isolation of the agent associated with the presence of typical clinical signs and pathological conditions. The aim of this study was to describe the development of a specific polymerase chain reaction (PCR) test for Conidiobolus lamprauges to detect the fungus in clinical samples. Samples from suspected animals were collected and submitted to isolation, histopathological analysis and amplification by PCR. DNA from tissues was subjected to PCR with fungi universal primers 18S rDNA gene, and specific primers were designed based on the same gene in C. lamprauges that generated products of about 540 bp and 222 bp respectively. The culture was positive in 26.6% of clinical samples. The PCR technique for C. lamprauges showed amplification of DNA from fresh tissues (80%) and paraffin sections (44.4%). In conclusion, the PCR technique described here demonstrated a high sensitivity and specificity for detection of fungal DNA in tissue samples, providing a tool for the rapid diagnosis of C. lamprauges.

Development of a Real-time PCR test for porcine group A rotavirus diagnosis

Group A Rotavirus (RVA) is one of the most common causes of diarrhea in humans and several animal species. A SYBR-Green Real-Time polymerase chain reaction (PCR) was developed to diagnose RVA from porcine fecal samples, targeting amplification of a 137-bp fragment of nonstructural protein 5 (NSP5) gene using mRNA of bovine NADH-desidrogenase-5 as exogenous internal control. Sixty-five samples were tested (25 tested positive for conventional PCR and genetic sequencing). The overall agreement (kappa) was 0.843, indicating 'very good' concordance between tests, presenting 100% of relative sensitivity (25+ Real Time PCR/25+ Conventional PCR) and 87.5% of relative sensitivity (35- Real Time PCR/40- Conventional PCR). The results also demonstrated high intra- and inter-assay reproducibility (coefficient of variation ≤1.42%); thus, this method proved to be a fast and sensitive approach for the diagnosis of RVA in pigs.

A survey of the generation of ocean waves in a test basin

At present stage the analytical design of wave tolerance for floating structures and vessels is still imperfect due to the mutually complex and nonlinear phenomena between structures and waves. Wave tolerance design is usually carried out through iterative evaluations of results from model tests in a wave basin, and this is done in order to reach a final structural design. The wave generation has then become an important technology in the field of the coastal and ocean engineering. This paper summarizes the facilities of a test basin and a wave maker in Japan and also surveys the methodology of the generation of ocean waves in a test basin.

A Test Bed Applied Mobile Robot Navigation

It is presented a test bed applied to studies on dynamics, control, and navigation of mobile robots. A cargo ship scale model was chosen, which can be radio-controlled or operated autonomously through an embedded control system. A control program, which manages on board mission execution, is implemented on a microcontroller. Navigation is based on an electronic compass, which includes automatic compensation for pitch and roll motions. Heading control loop is based on this sensor, and on a rudder positioning system. A propulsion control system is also implemented. Typical manoeuvres as the turning test and "zig-zag", were implemented and tested. They are included on a manoeuvre library, and can be accessed independently or in combined modes. The embedded system is also in charge of signal acquisition and storing during the missions. It is possible to analyse experiments on identification of ship dynamics, control, and navigation, through the data transferred to a PC by serial communication. Navigation is going to be improved by including inertial sensors on board, and a DGPS. Preliminary tests are aimed to ship identification, and manoeuvrability, using free model tests. Future steps include extending this system for developing other mobile robots as, ROVs, AUVs, and aerial vehicles.

Design of a Test Stand for a Centrifugal Compressor

The goal of the Master’s thesis is to design a test stand for a centrifugal compressor. Different theoretical aspects of flow parameters measurements and test rigs built for the similar purposes in other research units are described in the theoretical part of the work. The process of components selection and the description of chosen components are given in the second part of the thesis. Besides measuring and control stages, the designed test stand has a closed-loop piping, an aftercooler and a surge tank. Overview and layout of the test rig is presented in the last chapter of the work.

Preparation for ramp-up production process in metal manufacturin

Collaboration is essential for successful new product development. In the preparation for ramp-up production collaboration between R&D and supply chain functions is crucial. This thesis examines the meaning of collaboration and the effects of collaboration between R&D and supply chain. The aim of this thesis is to analyse and advice on how to improve the collaboration between the research and development department and supply chain within the preparation for rampup process. This thesis begins by introducing the reader to the product development methodologies and collaboration literature. The following part of the thesis describes the current situation and the results of the qualitative research. The last part of the thesis will explain the improvement suggestions. The main improvement suggestions are clarification of the processes and responsibilities and the introduction of a kick-off meeting.