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Present dataset aims at the inventory data on preparation of Desertification Status Maps (DSM) for the first time in semi-arid region of Anantapur district in the state of Andhra Pradesh, South India by applying Normalized Difference Vegetation Index (NDVI) with acquired Remote Sensing (RS) satellite imageries and processed in ERDAS Imagine and ArcGIS software's. The NDVI has been classified into five such as water body, vegetation, fallow land, degradation land, and desertified land. Further, degradation land has been decreased to 4.87% which lead to desertification in the study region. The current research data will be resourceful to the environmental scientists and planning agencies who can utilize optimum for sustainable development and good governance in land degradation, desertification, and conserve land resources.
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BACKGROUND AND AIMS: Laryngoscopy forms an important part of general anesthesia and endotracheal intubation. The aim of the present study was to compare the hemodynamic responses to Laryngoscopy and Intubation using Macintosh or McCoy or C-MAC Laryngoscope with M-Entropy module monitoring to ensure uniform and adequate depth of anesthesia, during and after intubation. MATERIAL AND METHODS: A prospective, randomised, comparative study was done and patients included were of 18 to 60 years, ASA (American Society of Anesthesiologist) physical status I and II of both sexes undergoing elective surgery under general anesthesia. They were assigned to three groups using simple randomisation, after securing IV (intravenous) access, standard monitoring and Entropy leads were attached. General anesthesia was administered with glycopyrrolate 0.1 mg, fentanyl 2 ug/kg and intravenous thiopentone, 4 mg/kg. Adequate muscle relaxation was achieved with atracurium 0.6 mg/kg IV. By titrating isoflurane concentration, Entropy maintained between 40 and 60, orotracheal intubation done, with Macintosh or McCoy or C-MAC blades according to simple randomisation. Size of laryngoscope blade, time taken for laryngoscopy and intubation were noted. Heart rate, blood pressure, RE (Response Entropy) and SE (State Entropy) were noted before and during induction and laryngoscopy and post intubation up to 5 minutes. Statistical analysis done using NCSS 9 version 9.0.8 statistical software. RESULTS: Hemodynamic responses during laryngoscopy and intubation using Macintosh or McCoy or C-MAC laryngoscope were statistically insignificant (p > 0.05) between the three groups, provided the depth of anesthesia is maintained constant. CONCLUSIONS: It is the depth of anesthesia that decides the magnitude of hemodynamic responses and not the choice of laryngoscope.
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The data deals with the functions that automatically extracted lineaments from the Cartosat, ASTER and SRTM of Digital Elevation Model (DEM) of different spatial resolutions, in the software ArcGIS 10.4. The extracted lineaments result shows the ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) DEM gives the lowest number of lineaments reflects Cartosat and SRTM (Shuttle Radar Topography Mission) DEM shows a medium number of lineaments. Cartosat DEM is most appropriate for extraction of contours precisely rather than ASTER and SRTM. This study reveals the Cartosat DEM data is best to use extraction of lineaments in the Indian provinces, offers at most comprehensive geological structural info amongst all the data sets. The extracted lineaments lengths and densities are determined by the statistical method. Based on the data generated lineament density and rose diagram. Cartosat DEM data are the best suited for studying very small areas as through geological and structural information can be mined by using this data.
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The Geospatial Technologies like Remote Sensing (RS) and Geographic Information System (GIS) have been playing vital role in capable forecasting and management of imperative groundwater resources in the emerging nations. In recent times, the geospatial technologies like RS, GIS and Multi Influence Factor (MIF) methodology are helpful in identifying groundwater potential zone. For the present study, the geospatial technology is used to prepare various thematic maps such as Land Slope, Geomorphology, Geology, Soil, Drainage Density, Lineament Density, Landuse/Landcover, Hydrogeomorphology, and Annual consideration of the valuation of groundwater assets for the semi-arid region in and around Bommanahal Mandal of Anantapur District in Andhra Pradesh, Southern India. As a part of the study eight thematic layers and their functions have been designed applicable weights at the Saaty׳s scale according to their comparative connotation in groundwater occurrence. The designed weights are normalized by using AHP (Analytic Hierarchy Process) MIF techniques and eigenvector method to various thematic layers and their features. Further to create a groundwater potential map the chosen thematic maps are integrated by weighted linear grouping method in a GIS environment. Based on the groundwater potential index values, the study area is classified into four different groundwater potential zones such as 'good', 'moderate to good', 'moderate' and 'poor'. The new recharge structures have proposed to fulfill the demand of groundwater to expand the scope of groundwater for future generations. Considering the overlay analysis of geomorphology and drainage layer execution through GIS technologies, the appropriate sites for artificial recharge structures have been identified.
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We report an unusual balanced translocation involving chromosomes 4 and 21 in a lady who had Down syndrome in her previous child. The most plausible explanation for this event is the 3:1 segregation of chromosomes at meiosis in her gametes leading to interchange trisomy 21.
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Segregación Cromosómica , Cromosomas Humanos Par 4 , Síndrome de Down/genética , Translocación Genética/genética , Adulto , Femenino , Asesoramiento Genético , HumanosRESUMEN
We used thiourea-induced thyroid hormone depletion as a strategy to understand the influence of thyroid hormones on testicular recrudescence of the air-breathing catfish, Clarias gariepinus. Treatment with 0.03% thiourea via immersion for 21 days induced hypothyroidism (thyroid hormone depletion) as evidenced by significantly reduced serum T(3) levels. Thiourea-treated males had narrowed seminiferous lobules with fewer spermatozoa in testis, very little or no secretory fluid, reduced protein and sialic acid levels in seminal vesicles when compared to controls. The histological changes were accompanied by reduction in serum and tissue levels of testosterone (T) and 11-ketotestosterone (11-KT), a potent male specific androgen in fish. Qualitative changes in the localization of catfish gonadotropin-releasing hormone (cfGnRH) and luteinizing hormone (LH, heterologous system) revealed a reduction in the distribution of immunoreactive neuronal cells and fibers in thyroid depleted fish. Interestingly, thiourea-withdrawal group showed physiological and histological signs of recovery after 21 days such as reappearance of spermatozoa and partial restoration of 11-KT and T levels. These data demonstrate that thyroid hormones play a significant role in testicular function of catfish. The mechanism of action includes modulating sex steroids either directly or through the hypothalamo (GnRH)-hypophyseal (LH) axis.
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Bagres/metabolismo , Testículo/efectos de los fármacos , Tiourea/farmacología , Hormonas Tiroideas/metabolismo , Animales , Hormona Liberadora de Gonadotropina/efectos de los fármacos , Hormona Liberadora de Gonadotropina/metabolismo , Hormona Luteinizante/efectos de los fármacos , Hormona Luteinizante/metabolismo , Masculino , Ácido N-Acetilneuramínico/metabolismo , Estaciones del Año , Vesículas Seminales/efectos de los fármacos , Vesículas Seminales/fisiopatología , Especificidad de la Especie , Testículo/fisiopatología , Testosterona/análogos & derivados , Testosterona/sangre , Testosterona/metabolismo , Triyodotironina/sangre , Triyodotironina/efectos de los fármacosRESUMEN
Rinderpest causes a devastating disease, often fatal, in wild and domestic ruminants. It has been eradicated successfully using a live, attenuated vaccine from most part of the world leaving a few foci of disease in parts of Africa, the Middle East, and South Asia. We have developed transgenic peanut (Arachis hypogaea L.) plants expressing hemagglutinin (H) protein of rinderpest virus (RPV), which is antigenically authentic. In this work, we have evaluated the immunogenicity of peanut-expressed H protein using mouse model, administered parenterally as well as orally. Intraperitoneal immunization of mice with the transgenic peanut extract elicited antibody response specific to H. These antibodies neutralized virus infectivity in vitro. Oral immunization of mice with transgenic peanut induced H-specific serum IgG and IgA antibodies. The systemic and oral immunogenicity of plant-derived H in absence of any adjuvant indicates the potential of edible vaccine for rinderpest.
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Arachis/genética , Glicoproteínas/administración & dosificación , Glicoproteínas/inmunología , Plantas Modificadas Genéticamente , Peste Bovina/prevención & control , Proteínas Virales/administración & dosificación , Proteínas Virales/inmunología , Vacunas Virales/inmunología , Administración Oral , Animales , Anticuerpos Antivirales/sangre , Modelos Animales de Enfermedad , Femenino , Glicoproteínas/genética , Hemaglutininas Virales , Inyecciones Intraperitoneales , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Pruebas de Neutralización , Virus de la Peste Bovina/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Proteínas Virales/genética , Vacunas Virales/administración & dosificación , Vacunas Virales/genéticaRESUMEN
A competitive enzyme-linked immunosorbent assay (C-ELISA) which detects antibodies unique to rinderpest virus (RPV) has been developed. This test can differentiate antibodies against RPV and those against peste des petits ruminants virus. The recombinant RPV hemagglutinin (H)-protein C-ELISA (recH C-ELISA) is based on the ability of a well-characterized monoclonal antibody (MAb) produced with the soluble, secreted form of the H protein (Sec H protein) of RPV made in a baculovirus expression system to compete with the binding of RPV antibodies in the serum of vaccinated or infected, recovered animals to the Sec H protein. The B-cell epitope recognized by the MAb corresponds to amino acids 575 to 583 on the H protein, which is not present on the antigenically closely related peste des petits ruminants virus hemagglutinin-neuraminidase protein. Initially, a positive-negative threshold cutoff value for percent inhibition of 34 was established with 500 known RPV-negative serum samples. The recH C-ELISA was developed with the enzyme immunoassay software of a commercial RPV C-ELISA kit. Comparative analysis of the test results for 700 serum samples obtained with the commercial kit gave a sensitivity of 112.4% and a specificity of 72.4%. Variations in percent inhibition values were observed for the two assay systems. These variations may have been due to the undefined amount of antigen present in the commercial kit as well as the use of a different MAb. The recH C-ELISA detected more positive serum samples compared to the number detected by the commercial kit, with the results confirmed by a virus neutralization test. Thus, recH C-ELISA is a sensitive tool for RPV serosurveillance in disease eradication programs.
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Ensayo de Inmunoadsorción Enzimática/métodos , Glicoproteínas/análisis , Virus de la Peste Bovina/aislamiento & purificación , Proteínas Virales/análisis , Animales , Anticuerpos Monoclonales/inmunología , Bovinos , Células Cultivadas , Glicoproteínas/inmunología , Hemaglutininas/inmunología , Hemaglutininas Virales , Juego de Reactivos para Diagnóstico , Proteínas Recombinantes/inmunología , Virus de la Peste Bovina/química , Virus de la Peste Bovina/inmunología , Spodoptera , Proteínas Virales/inmunologíaRESUMEN
Different doses of ethanolic fraction of fruits of Terminalia pallida were evaluated for hypoglycemic and antihyperglycemic activity in normal and alloxan diabetic rats. The oral administration of ethanolic extract at a dosage of 0.5 g/kg body weight exhibited a significant antihyperglycemic activity in alloxan diabetic rats, whereas in normal rats no hypoglycemic activity was observed.
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Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Fitoterapia , Terminalia , Administración Oral , Animales , Glucemia/análisis , Relación Dosis-Respuesta a Droga , Etanol , Frutas , Masculino , Extractos Vegetales/uso terapéutico , Ratas , Ratas WistarRESUMEN
In India, brucellosis was first recognised in 1942 and is now endemic throughout the country. The disease is reported in cattle, buffalo, sheep, goats, pigs, dogs and humans. B. abortus biotype-1 in cattle and buffaloes and B. melitensis biotype-1 in sheep, goats and man are the predominant infective biotypes. The long-term serological studies have indicated that 5% of cattle and 3% of buffaloes are infected with brucellosis. Economic losses due to brucellosis in livestock are considerable in an agrarian country like India. There is no organised and effective brucellosis control programme in the country. With the indigenous development of serum and milk based ELISA kits, the population survey of the disease has been undertaken on a large scale in several states and plans for the control of the disease through calf-hood vaccination are being worked out. An innovative approach--Bovine Brucellosis Progressive Control Programme (BBPCP) is targeted to overcome the basic problems of ban on cow slaughter, distress sale of animals following the positive serological diagnosis of brucellosis and absence of a disease control strategy. The work plan for the implementation of BBPCP is presented.
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Brucelosis/veterinaria , Vacunación/veterinaria , Animales , Animales Domésticos , Brucella abortus/clasificación , Brucella melitensis/clasificación , Brucelosis/diagnóstico , Brucelosis/epidemiología , Brucelosis/prevención & control , Brucelosis Bovina/diagnóstico , Brucelosis Bovina/epidemiología , Brucelosis Bovina/microbiología , Brucelosis Bovina/prevención & control , Bovinos , Ensayo de Inmunoadsorción Enzimática , Geografía , Humanos , India/epidemiología , Prevalencia , Juego de Reactivos para Diagnóstico , Vacunación/métodos , Zoonosis/epidemiologíaRESUMEN
A recombinant baculovirus expressing membrane bound form of hemagglutinin-neuraminidase (HN) protein of peste des petits ruminants virus (PPRV) was employed to generate monoclonal antibodies (mAbs) against PPRV-HN protein. Four different mAbs were employed for mapping of regions on HN carrying B-cell epitopes using deletion mutants of PPRV-HN and RPV-H proteins expressed in Escherichia coli as well as PPRV-HN deletion proteins expressed transiently in mammalian cells. The immuno-reactivity pattern indicated that all mAbs bind to two discontinuous regions of amino acid sequence 263-368 and 538-609 and hence the epitopes identified are conformation-dependent. The binding regions for three mAbs were shown to be immunodominant employing competitive ELISA with vaccinated sheep sera. Delineation of functional domains on PPRV-HN was carried out by assessing the ability of these mAbs to inhibit neuramindase activity and hemagglutination activity. Two mAbs inhibited NA activity by more than 63% with substrate N-acetyl neuraminolactose, while with Fetuin one mAb showed inhibition of NA activity (95%). Of the three antigenic sites identified based on competitive inhibition assay, site 2 could be antigenically separated into 2a and 2b based on inhibition properties. All the four mAbs are virus neutralizing and recognized PPRV-HN in immunofluorescence assay.
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Anticuerpos Monoclonales/inmunología , Mapeo Epitopo , Epítopos de Linfocito B/inmunología , Proteína HN/química , Proteína HN/metabolismo , Virus de la Peste de los Pequeños Rumiantes/inmunología , Animales , Linfocitos B/inmunología , Células Cultivadas , Pollos , Chlorocebus aethiops , Eliminación de Gen , Proteína HN/inmunología , Pruebas de Inhibición de Hemaglutinación , Pruebas de Hemaglutinación , Hibridomas , Epítopos Inmunodominantes , Conformación Proteica , Spodoptera , Células VeroRESUMEN
Monoclonal antibodies (mAbs) against secreted hemagglutinin (H) protein of rinderpest virus (RPV) expressed by a recombinant baculovirus were generated to characterize the antigenic sites on H protein and regions of functional significance. Three of the mAbs displayed hemagglutination inhibition activity and these mAbs were unable to neutralize virus infectivity. Western immunoblot analysis of overlapping deletion mutants indicated that three mAbs recognize antigenic regions at the extreme carboxy terminus (between amino acids 569 and 609) and the fourth mAb between amino acids 512 and 568. Using synthetic peptides, aa 569-577 and 575-583 were identified as the epitopes for E2G4 and D2F4, respectively. The epitopic domains of A12A9 and E2B6 mAbs were mapped to regions encompassing aa 527-554 and 588-609. Two epitopes spanning the extreme carboxy terminal region of aa 573 to 587 and 588 to 609 were shown to be immunodominant employing a competitive ELISA with polyclonal sera form vaccinated cattle. The D2F4 mAb which recognizes a unique epitope on RPV-H is not present on the closely related peste des petits ruminant virus HN protein and this mAb could serve as a tool in the seromonitoring program after rinderpest vaccination.
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Epítopos de Linfocito B/inmunología , Glicoproteínas/inmunología , Virus de la Peste Bovina/inmunología , Proteínas Virales/inmunología , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/inmunología , Bovinos , Mapeo Epitopo , Eliminación de Gen , Glicoproteínas/genética , Hemaglutininas Virales , Sueros Inmunes , Ratones , Ratones Endogámicos BALB C , Pruebas de Neutralización , Péptidos/síntesis química , Péptidos/inmunología , Proteínas Recombinantes/inmunología , Proteínas Virales/genéticaRESUMEN
Because previous authorities had suggested that small ruminants were playing a part in the dissemination of rinderpest, and a rinderpest-eradication campaign was about to begin, it was necessary to make precise virus identifications from a number of small-ruminant "rinderpest" outbreaks. When this was done using a database created from passive disease reports, we found that epidemics-reportedly due to rinderpest-were in fact due to peste des petits ruminants (PPRs). Although such cases had been common in India for a number of years, earlier clinical and laboratory reports no longer should be regarded as definitive. PPR outbreaks have been frequent in recent years. Further, we suggest that PPR is not a recent invader of India.
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Brotes de Enfermedades/veterinaria , Peste de los Pequeños Rumiantes/epidemiología , Virus de la Peste de los Pequeños Rumiantes/aislamiento & purificación , Animales , Cabras , India/epidemiología , Estaciones del Año , OvinosRESUMEN
The avidin-biotin enzyme-linked immunosorbent assay (A-B ELISA), for use in surveillance for bovine brucellosis in India was developed and calibrated using the indirect brucellosis ELISA kit of the International Atomic Energy Agency (IAEA) as a reference. The reagents used in the A-B ELISA were as follows: the smooth lipopolysaccharide of Brucella abortus strain 99 (antigen); biotinylated anti-bovine immunoglobulin G (detection antibody); avidin-horseradish peroxidase (conjugate); and O-phenylenediamine dihydrochloride (chromogen). The test results were interpreted using the IAEA software EDI version 2.1.1, which was modified for use in the A-B ELISA. The cut-off percentage positivity value was established using 500 brucellosis-positive and 500 brucellosis-negative serum samples, confirmed with reference to the sample data using the indirect ELISA kit. The overall specificity of A-B ELISA was 98.8% and overall sensitivity was 98.2%. Field validation of the A-B ELISA kit was undertaken in six laboratories in India. Screening of 7,040 cattle and 678 buffalo serum samples from 12 states revealed serological evidence of brucellosis in 8.7% of cattle and 10.2% of buffalo. This kit proved to be robust and performed with a similar sensitivity and specificity to the indirect ELISA. The kit can be supplied at a lower cost than current commercial ELISA kits.
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Anticuerpos Antibacterianos/sangre , Brucella abortus/inmunología , Brucelosis Bovina/diagnóstico , Búfalos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Animales , Avidina , Biotina , Brucelosis Bovina/epidemiología , Bovinos , Sueros Inmunes/inmunología , Lipopolisacáridos/inmunologíaRESUMEN
Peste des petits ruminants virus (PPRV), a member of the genus Morbillivirus within the family Paramyxoviridae, causes a fatal disease 'peste des petits ruminants' in goats and sheep. This enveloped virus is antigenically closely related to rinderpest virus (RPV), which causes a similar but distinct disease in large ruminants. PPRV harbors two major surface glycoproteins, the hemagglutinin-neuraminidase (HN) and the fusion (F) proteins. The surface glycoproteins of morbilliviruses are highly immunogenic and confer protective immunity. In this study, we investigated the immune responses generated in goats immunized with low doses of purified recombinant extracellular baculovirus carrying a membrane bound form of the HN protein of PPRV without any adjuvant. We report that the immunized goats develop both humoral and cell-mediated immune responses. Antibodies generated in the immunized animals could neutralize both PPRV and RPV in vitro. Further, using a combination of Escherichia coli expressed deletion mutants of PPRV-HN and RPV-H proteins, and synthetic peptides corresponding to the highly conserved N-terminal sequences of MV-H protein, we have mapped an N-terminal T cell determinant (amino acids 123-137) and a C-terminal domain (amino acids 242-609) harboring potential T cell determinant(s) in goats.
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Antígenos Virales/inmunología , Enfermedades de las Cabras/prevención & control , Cabras/inmunología , Proteína HN/inmunología , Peste de los Pequeños Rumiantes/prevención & control , Virus de la Peste de los Pequeños Rumiantes/inmunología , Subgrupos de Linfocitos T/inmunología , Vacunación/veterinaria , Secuencia de Aminoácidos , Animales , Anticuerpos Antivirales/biosíntesis , Anticuerpos Antivirales/inmunología , Antígenos Virales/genética , Reacciones Cruzadas , ADN Complementario/genética , Ensayo de Inmunoadsorción Enzimática , Escherichia coli , Glicoproteínas/genética , Glicoproteínas/inmunología , Enfermedades de las Cabras/inmunología , Proteína HN/genética , Inmunidad Celular , Activación de Linfocitos , Proteínas de la Membrana , Datos de Secuencia Molecular , Peste de los Pequeños Rumiantes/inmunología , Proteínas Recombinantes/inmunología , Virus de la Peste Bovina/inmunología , Eliminación de Secuencia , Proteínas Virales de Fusión/genética , Proteínas Virales de Fusión/inmunologíaRESUMEN
Rinderpest virus causes a highly contagious and often fatal disease in domestic and wild ruminants. The surface glycoproteins, hemagglutinin (H) and fusion (F) proteins of this enveloped virus are known to confer protective immunity in cattle. We have reported the generation of a recombinant baculovirus expressing H protein and studied its protective properties in cattle. In this report, we demonstrate that the recombinant baculovirus encoded H protein expressed in insect cells gets incorporated into extracellular baculovirus. Single administration of low doses of purified recombinant extracellular virus with or without adjuvant induces virus neutralizing antibody responses and bovine leukocyte antigen (BoLA) class II restricted helper T cell responses in cattle.
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Glicoproteínas/inmunología , Virus de la Peste Bovina/inmunología , Proteínas Virales/inmunología , Animales , Anticuerpos Antivirales/biosíntesis , Baculoviridae/genética , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/prevención & control , Línea Celular , Glicoproteínas/genética , Hemaglutininas Virales , Antígenos de Histocompatibilidad Clase II/metabolismo , Inmunidad Celular , Activación de Linfocitos , Pruebas de Neutralización , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Peste Bovina/inmunología , Peste Bovina/prevención & control , Virus de la Peste Bovina/genética , Spodoptera , Linfocitos T Colaboradores-Inductores/inmunología , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/farmacología , Proteínas Virales/genética , Vacunas Virales/genética , Vacunas Virales/inmunología , Vacunas Virales/farmacologíaRESUMEN
Rinderpest virus (RPV), a member of the genus Morbillivirus within the Paramyxoviridae family, causes a highly contagious and often fatal disease known as rinderpest in wild and domestic ruminants. The envelope of the virus contains two surface glycoproteins, namely the hemagglutinin (H) and the fusion (F) proteins, both of which have been shown to confer protective immunity in animals. In this paper, we demonstrate that single administration of low doses of recombinant H protein of RPV expressed in insect cells in the form of extracellular virus induces long lasting bovine leukocyte antigen class I restricted cytotoxic T-cell (CTL) responses in cattle in the absence of adjuvant. This is the first report of CTL responses in cattle against one of the protective antigens of RPV.
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Bovinos/inmunología , Citotoxicidad Inmunológica , Glicoproteínas/inmunología , Virus de la Peste Bovina/inmunología , Linfocitos T/inmunología , Proteínas Virales/inmunología , Vacunas Virales/inmunología , Animales , Baculoviridae/genética , Células Cultivadas , Glicoproteínas/administración & dosificación , Glicoproteínas/biosíntesis , Hemaglutininas Virales , Antígenos de Histocompatibilidad Clase I/inmunología , Proteínas Recombinantes/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Proteínas Virales/administración & dosificación , Proteínas Virales/biosíntesis , Vacunas Virales/administración & dosificaciónRESUMEN
A multiplex amplification and detection platform for the diagnosis of Mycobacterium bovis and Brucella abortus infection simultaneously in bovine milk and nasal secretions was developed. This system (designated the bovine pathogen detection assay [BPDA]-PCR) consists of duplex amplification of species-specific targets (a region of the BCSP31K gene of B. abortus and a repeat-sequence region in the hsp65 gene of M. bovis, respectively). This is followed by a solid-phase probe capture hybridization of amplicons for detection. On the basis of spiking experiments with normal milk, the analytical sensitivity of the assay was 800 CFU equivalents/ml of milk for B. abortus and as low as 4 CFU equivalents per ml of milk for M. bovis. BPDA-PCR was validated with 45 liver samples from lemmings experimentally infected with B. abortus. The assay sensitivity, based on culture status as a "gold standard," was 93.9%. In this experiment, BPDA-PCR also identified five culture-negative liver samples as positive (41.7%). Field studies for the evaluation of BPDA-PCR were performed with samples from dairy animals from geographically distinct regions (India, Mexico, and Argentina). A high prevalence of shedding of B. abortus (samples from India) and M. bovis (samples from Mexico) was identified by BPDA-PCR. In samples from India, B. abortus shedding was identified in 86% of milk ring test-positive animals (n = 15) and 80% of milk ring test-negative cows (n = 5). In samples from Mexico, M. bovis was identified by PCR in 32.6% of pools (n = 46) of milk that each contained milk from 10 animals and in 56.2% of nasal swabs (n = 121) from cattle from tuberculin test-positive herds. In contrast, the Argentine cattle (n = 70) had a modest prevalence of M. bovis shedding in nasal swabs (2.9%) and milk (1.4%) and of B. abortus in milk (11.4%). On the basis of these analyses, we identify BPDA-PCR as an optimal tool for both screening of herds and testing of individual animals in a disease eradication program. A combination of the duplex assay, screening of milk samples in pools, and the proposed algorithm provides a highly sensitive, cost-effective, and economically viable alternative to serological testing.
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Brucella abortus/aislamiento & purificación , Brucelosis Bovina/diagnóstico , Mycobacterium bovis/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Tuberculosis Bovina/diagnóstico , Animales , Proteínas Bacterianas/genética , Brucella abortus/genética , Brucelosis Bovina/microbiología , Bovinos , Chaperonina 60 , Chaperoninas/genética , ADN Bacteriano/genética , Leche/microbiología , Mycobacterium bovis/genética , Cavidad Nasal/microbiología , Sensibilidad y Especificidad , Tuberculosis Bovina/microbiologíaRESUMEN
Isolation of Leptospira from the kidneys of Rattus rattus wroughtoni hinton, Rattus rattus rufescens, Bandicota bengalensis and Bandicota indica was attempted in Bangalore in southern India. In total, 296 spirochaetes were isolated from 1,348 kidney cultures (an isolation rate of 22%). A batch of fifty-six isolates from India was identified, based on serological and polymerase chain reaction analysis, of which twenty-three isolates were identified as L. inadai by the World Health Organization/Food and Agriculture Organization Collaborating Centre for Reference and Research on Leptospirosis, in Brisbane. This is the first record of isolation of L. inadai from rodents. The preponderance of L. inadai in four different species of rodents suggests that these animals could be the natural reservoir hosts of L. inadai, and raises a critical question as to the likely impact of this species of Leptospira on the renal carrier status of other Leptospira pathogenic to humans and animals in this part of India. Virulence studies conducted at the University of Trieste in Italy, revealed that isolates of L. inadai from India were moderately or totally serum resistant when subjected to a serum killing test. To establish the possible seroprevalence of this species in the population, the inclusion of L. inadai in the battery of leptospiral antigens used for sero-epidemiological studies is recommended.