Bovine monocytes and a macrophage cell line differ in their ability to phagocytose and support the intracellular survival of Mycobacterium avium subsp. paratuberculosis

Bovine monocytes exhibited a greater ability to phagocytose Mycobacterium avium subsp. paratuberculosis (i.e. greater percentage of infected cells, and more bacilli per infected cell), than did a bovine macrophage cell line (BoMac). Phagocytosis of M. paratuberculosis by monocytes, but not the cell line, was significantly enhanced by the addition of autologous serum. Following ingestion, the numbers of viable M. paratuberculosis cells in monocytes increased during the first 4 days and then declined between day 4 and day 8 after infection, as determined by a radiometric method. In contrast, BoMac cells were not permissive for bacillary multiplication; the numbers of M. paratuberculosis remained largely unchanged in the cell line during the 8 day incubation period. The numbers of microscopically visible acid-fast bacilli increased with time in monocytes but not in the macrophage cell line. These observations suggest that replication and inactivation of bacilli may both occur in monocytes. The differing abilities of bovine monocytes and the macrophage cell line to ingest and restrain the intracellular growth of M. paratuberculosis provide contrasting model systems for investigating how M. paratuberculosis enters and persists within its preferred niche, the mononuclear phagocyte.     

ATP release by infected bovine monocytes increases the intracellular survival of Mycobacterium avium subsp. paratuberculosis

Mycobacterium avium subsp. paratuberculosis is the etiologic agent of Johne's disease, a chronic intestinal infection in ruminants. Adenosine 5'-Triphosphate (ATP) has been reported to induce killing of several Mycobacterium species in human and murine macrophages. We investigated whether ATP secreted from M. avium subsp. paratuberculosis-infected bovine monocytes affects intracellular survival of the bacilli. Bovine monocytes constitutively secreted ATP during an 8-day incubation period in vitro; however, M. avium subsp. paratuberculosis infection did not enhance ATP release. Removal of extracellular ATP by the addition of apyrase increased the viability of infected monocytes, but surprisingly decreased the number of viable intracellular bacilli. In contrast to previous reports, addition of extracellular ATP (1mM) increased intracellular survival of M. avium subsp. paratuberculosis in bovine monocytes. Neither apyrase nor ATP altered production of reactive oxygen intermediates (ROI) or reactive nitrogen intermediates (RNI) by bovine monocytes. These results suggest that ATP release from infected bovine monocytes improves, rather than decreases, the intracellular survival of M. avium subsp. paratuberculosis.     

Rapid Identification of Clavibacter michiganensis Subspecies sepedonicus Using Two Primers Random Amplified Polymorphic DNA (TP-RAPD) Fingerprints

Twenty strains of Clavibacter michiganensis subsp. sepedonicus from different geographic origins and other reference strains of the same and different species, including other potato pathogens, were analysed with a new procedure named TP-RAPD that originates fingerprints of bacterial species. This procedure uses two primers to amplify the 16S rDNA gene. At 45 °C of annealing, the PCR product electrophoresed in agarose gels produced a band pattern that was different in all bacterial species studied as well as in the subspecies of C. michiganensis. All strains of C. michiganensis subsp. sepedonicus displayed the same TP-RAPD number of pattern. Unlike Gram negative bacteria, Gram positives of high G + C content, such as Clavibacter, produced low bands in TP-RAPD. By using a different set of two primers also based in the 16S rDNA sequence from Escherichia coli a more adequate amplification of Gram positives of high G + C including a greater number of bands was obtained. TP-RAPD patterns using the new set of primers described in this work is a reliable and fast method to identify C. michiganensis subsp. sepedonicus.     

Authentication of Rice Using Near‐Infrared Reflectance Spectroscopy

The authentication of rice (Korean domestic rice vs. foreign rice) has been attempted using near‐infrared spectroscopy (NIRS). Two sample sets (n₁ = 280 and n₂ = 200) were used to obtain calibration equations and the spectral regions used for this study were 500–600 nm, 700–900nm, and 980–2,498 nm. Modified partial least square (MPLS) regression was used to develop the prediction model. The standard error of cross validation (SECV) and the r² were 0.165 and 0.91 respectively for 1st calibration set and 0.165 and 0.93 for 2nd calibration set respectively. The results of the independent validation (n₃ = 80) showed that all of 80 samples were identified correctly. Even though authentication of rice was performed successfully using NIRS, the calibration statistics in this study showed that further effort is needed for implementation of NIRS for authentication of rice for industry purposes.     

Ancient DNA evidence for Old World origin of New World dogs

Mitochondrial DNA sequences isolated from ancient dog remains from Latin America and Alaska showed that native American dogs originated from multiple Old World lineages of dogs that accompanied late Pleistocene humans across the Bering Strait. One clade of dog sequences was unique to the New World, which is consistent with a period of geographic isolation. This unique clade was absent from a large sample of modern dogs, which implies that European colonists systematically discouraged the breeding of native American dogs.     

Behavior of Impatiens wallerana Hook. F in Alternative Pot Substrates: Mechanisms Involved and Research Perspectives

The approach to select new growing media, has been focused on selecting materials only from the physical point of view. The objective of this study was to describe the physiological mechanisms involved in I. wallerana growth when cropped on a broad range of growing media created from alternative components. Results showed a close relationship between I. wallerana growth and fine particle size at the beginning of the experiments. Shoot fresh weight was determined mainly by the root system size. There were small differences in the relative growth rate (RGR) between the control substrate and the thirty alternative substrates tested. The lower RGR values resulted from a decrease in the net assimilation rate and the leaf area ratio. The mechanism involved would be associated with a change in photosynthate partitioning, which favored root growth. A close relationship between growth (as total dry weight) and nitrogen content was found as well.     

Quantification of dwarfing effect of different rootstocks in ‘Picual’ olive cultivar using UAV-photogrammetry

Precision Agriculture - Hedgerow orchard is an olive growing system where trees are planted at a super high-density higher than 20-fold (i.e., 1200–2500 trees ha?1) compared to the...     

Association between major histocompatibility complex microsatellites, fecal egg count, blood packed cell volume and blood eosinophilia in Pelibuey sheep infected with Haemonchus contortus

The objective of this study was to assess the correlation among traits associated with resistance or susceptibility to Haemonchus contortus infestation and to evaluate the participation of the ovine major histocompatibility complex (MHC) in Pelibuey sheep, a prevalent breed in tropical and sub-tropical regions in Mexico and elsewhere. Association among the fecal egg count (FEC), blood packed cell volume (PCV), antibody (AB) levels, serum proteins (SP) and blood eosinophil count (EOS) was assessed in 52 lambs experimentally infected with H. contortus, and the participation of the MHC was evaluated using polymorphisms in three microsatellites, located at the class I (OMHC1) and class II (OLADRB1, OLADRB2) regions of the MHC. Spearman correlation analysis among the traits showed a negative association (P<0.01) between FEC and PCV (-0.35), EOS (-0.50), SP (-0.30) and AB (-0.57), and a positive correlation of antibodies with EOS (0.50). The homozygotes for the OMHC1-188 and OLADRB2-282 alleles were associated with a reduction in FEC (-813 and -551, respectively). Conversely, the OMHC1-200 and OMHC1-206 alleles were associated with an increase in FEC (1704 and 1008, respectively). Furthermore, the OLADRB1-482 allele was associated with an increase of 163 EOS by allele copy, while the OMHC1-200 allele showed a reduction of 95 EOS in homozygotes. The associations among microsatellite MHC loci and the remaining variables were not significant. These results reinforce the evidence that MHC polymorphisms have an important role in parasite resistance or susceptibility in Pelibuey sheep and could be used as genetic markers to assist selection and improve parasite resistance to H. contortus.     

Prevalence of Leishmania spp. infection in domestic dogs in Chapare, Bolivia

Data on Leishmania spp. infection in dogs in Bolivia is scarce. Dogs from an area where 90% of human cutaneous leishmaniasis (CL) cases are due to Leishmania (Viannia) braziliensis were screened for Leishmania infection using established enzyme-linked immunosorbent antibody test (ELISA) protocols. Although none of the 51 dogs surveyed had clinical lesions indicative of CL, 6 out of 51 (11.8%) sampled dogs tested positive by ELISA.