Application of a multiplex PCR assay for Campylobacter fetus detection and subspecies differentiation in uncultured samples of aborted bovine fetuses     

Gregorio Iraola  Martín Hernández  Lucía Calleros  Fernando Paolicchi  Silvia Silveyra  Alejandra Velilla  Luis Carretto  Eliana Rodríguez  Ruben Pérez 《Journal of veterinary science (Suw?n-si, Korea)》2012,13(4):371-376

Campylobacter (C.) fetus (epsilonproteobacteria) is an important veterinary pathogen. This species is currently divided into C. fetus subspecies (subsp.) fetus (Cff) and C. fetus subsp. venerealis (Cfv). Cfv is the causative agent of bovine genital Campylobacteriosis, an infectious disease that leads to severe reproductive problems in cattle worldwide. Cff is a more general pathogen that causes reproductive problems mainly in sheep although cattle can also be affected. Here we describe a multiplex PCR method to detect C. fetus and differentiate between subspecies in a single step. The assay was standardized using cultured strains and successfully used to analyze the abomasal liquid of aborted bovine fetuses without any pre-enrichment step. Results of our assay were completely consistent with those of traditional bacteriological diagnostic methods. Furthermore, the multiplex PCR technique we developed may be easily adopted by any molecular diagnostic laboratory as a complementary tool for detecting C. fetus subspecies and obtaining epidemiological information about abortion events in cattle.  相似文献   

Rapid methodology for antigenic profiling of FMDV field strains and for the control of identity, purity and viral integrity in commercial virus vaccines using monoclonal antibodies     

Seki C  Robiolo B  Periolo O  Iglesias M  D'Antuono A  Maradei E  Barros V  La Torre J  Mattion N 《Veterinary microbiology》2009,133(3):239-251

Monoclonal antibodies (MAbs) developed against different foot-and-mouth disease virus (FMDV) vaccine strains were extensively used to study any possible antigenic variations during vaccine production in Argentine facilities. Additionally, a typing ELISA using strain specific MAbs was developed to detect potential cross contaminations among FMDV strains in master and working seeds with high specificity and sensitivity and to confirm strains identity in formulated vaccines. This assay was carried out for the South American strains currently in use in production facilities in Argentina (A24/Cruzeiro, A/Argentina/01, O1/Campos and C3/Indaial) and for the strain O/Taiwan, produced only for export to Asia. These non-cross reactive MAbs were also used to analyze the integrity of viral particles belonging to each one of the individual strains, following isolation of 140S virions by means of sucrose density gradients from the aqueous phase of commercial polyvalent vaccines. Antigenic profiles were defined for FMDV reference strains using panels of MAbs, and a coefficient of correlation of reactivity with these panels was calculated to establish consistent identity upon serial passages of master and production seeds. A comparison of vaccine and field strain antigenic profiles performed using coefficients of correlation allowed the rapid identification of two main groups of serotype A viruses collected during the last FMD epidemic in Argentina, whose reactivity matched closely to A/Argentina/2000 and A/Argentina/2001 strains.  相似文献   

Amaranth (Amaranthus hypochondriacus) vicilin subunit structure     

Quiroga A  Martínez EN  Rogniaux H  Geairon A  Añón MC 《Journal of agricultural and food chemistry》2010,58(24):12957-12963

The 7S-globulin fraction is a minor component of the amaranth storage proteins. The present work provides new information about this protein. The amaranth 7S-globulin or vicilin presented a sedimentation coefficient of 8.6 ± 0.6 S and was composed of main subunits of 66, 52, 38, and 16 kDa. On the basis of mass spectrometry (MS) analysis of tryptic fragments, the 52, 38, and 16 kDa subunits presented sequence homology with sesame vicilin, whereas the 66 kDa subunit showed sequence similarity with a putative vicilin. Several characteristics of the 66 kDa subunit were similar to members of the convicilin family. Results support the hypothesis that the 7S-globulin molecules are composed of subunits coming from at least two gene families with primary products of 66 and 52 kDa, respectively. According to the present information, amaranth vicilin may be classified into the vicilin group that includes pea, broad bean, and sesame vicilins, among others.  相似文献   

Foreword: Integrated plant disease management     

Michael Jeger  Maria Ivone da Clara  Alan Phillips  Dale Walters  Piet Boonekamp 《European journal of plant pathology / European Foundation for Plant Pathology》2012,133(1):1

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Prolactin stimulates the internalization of Staphylococcus aureus and modulates the expression of inflammatory response genes in bovine mammary epithelial cells     

Gutiérrez-Barroso A  Anaya-López JL  Lara-Zárate L  Loeza-Lara PD  López-Meza JE  Ochoa-Zarzosa A 《Veterinary immunology and immunopathology》2008,121(1-2):113-122

The incidence of mastitis in dairy cattle is highest at the drying off period and parturition, which are characterized by high levels of the lactogenic hormone prolactin (PRL). One of the most frequently isolated contagious pathogens causing mastitis is Staphylococcus aureus. However, the role of PRL on S. aureus infection in mammary epithelium has not been studied. In this work we evaluated the effect of bovine PRL (bPRL) on S. aureus internalization in a primary culture of bovine mammary epithelial cells (bMEC) and on the expression of cytokine and innate immune response genes. Our data show that 5ng/mL bPRL enhances approximately 3-fold the internalization of S. aureus (ATCC 27543) into bMEC. By RT-PCR analysis, we showed that bPRL is able to up-regulate the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and inducible nitric oxide synthase (iNOS) mRNAs. However, bPRL together with S. aureus did not modify the expression of TNF-alpha and iNOS mRNAs, while it down-regulated the expression of beta-defensin and IL-1beta mRNAs, as well as nitric oxide production, suggesting that infection and bPRL together can inhibit elements of the host immune response. To our knowledge, this is the first report that shows a role of bPRL during the internalization of S. aureus into bMEC.  相似文献   

Temporal and quantitative analyses of stem lesion development and foliar disease progression of peach rust in california     

Soto-Estrada A  Adaskaveg JE 《Phytopathology》2004,94(1):52-60

ABSTRACT The development of rust epidemics caused by Tranzschelia discolor on leaves and stems of cling peach was studied in California orchards. Sporulating stems lesions were only detected from late March until July in 1997 and 1998. When rust was present in the fall, the quadratic equation Y = -82.51 + 1.97JD - 0.01JD(2) using Julian day (JD) described the incidence of sporulating lesions on stems of cv. Andross (R(2) = 0.73; P /=117.3 mm of total precipitation and maximum temperatures of 相似文献   

Effects of biotic and abiotic factors on δ15N in young Pinus radiata     

Alejandra Couto-Vázquez  Serafín J. González-Prieto 《European Journal of Forest Research》2014,133(4):631-637

In a 12-year-old Pinus radiata plantation, three dominant and three suppressed trees removed by thinning were randomly selected, and needles, annual rings from basal stem disks and bark were collected and analyzed to study the relationships of climate, tree age, dominance and growth with tree δ¹⁵N. The high foliar N concentration (1.35–2.73 % N, dw) suggested that N was not limiting tree growth, therefore allowing plants to fractionate versus δ¹⁵N, leading to differences in δ¹⁵N among trees. Most wood δ¹⁵N_air values were below the δ¹⁵N_air natural abundance in the dominant pines (?2.43 to +1.69 ‰) and above it in the suppressed trees (+0.73 to +3.35 ‰), likely due to the access of dominants to exogenous N sources with lower δ¹⁵N_air than those of suppressed. However, no dominance effect was detected in δ¹⁵N_air of bark and needles that decreased in the order: buds (+1.20 to +2.44 ‰) > needles 1 year (?0.27 to +1.43 ‰) > needles 2 years (?0.97 to +0.41 ‰) > bark (?1.18 to +0.15 ‰). Compared with the soil N in the 0–15 cm layer (δ¹⁵N_air = +4.8 ‰), all plant material was ¹⁵N-depleted. Results suggest that seedlings and foliar buds have a less efficient system for N conservation and recycling, with higher losses. The linear regression models showed that both biotic (dominance and tree age) and abiotic factors (temperature in spring–summer and annual precipitation) are needed to explain the wood δ¹⁵N_air satisfactorily.  相似文献   

Neospora caninum is a cause of perinatal mortality in axis deer (Axis axis)     

Walter Basso  Gastón Moré  Maria Alejandra Quiroga  Diego Balducchi  Gereon Schares  Maria Cecilia Venturini 《Veterinary parasitology》2014,199(3-4):255-258

Neospora caninum is a worldwide distributed protozoan that may cause neuromuscular disease in dogs and reproductive failure in domestic and wild ruminants. One axis fawn (Axis axis) and four neonates from the same deer herd died at a zoo in Argentina within a four-month period. The fawn presented with dilatation of the anal sphincter at birth and incontinence, developed weakness and ataxia and died at 14 days of age. At necropsy, a mega formation of the distal large intestine was observed. Microscopically, non-suppurative encephalitis, suppurative bronchopneumonia, fibrin necrotic enteritis and degenerative changes in the liver were observed in hematoxilin and eosin-stained tissue sections, and thick-walled N. caninum-like cysts were observed in fresh brain samples. Serologic studies for N. caninum revealed an IFAT titer of 1:6400 in the fawn and 1:25, 1:400, 1:3200 and 1:6400 in the neonates. N. caninum DNA was detected in brain samples from the fawn and from one neonate by PCR, and the parasite was isolated in vitro from the fawn’ brain after passage through gerbils (Meriones unguiculatus) and gamma-interferon knock-out mice. N. caninum DNA obtained from the fawn, neonate and isolated parasites showed the same microsatellite pattern. This suggests a common infection source for both animals.The diagnosis of N. caninum infection was confirmed, suggesting its association with perinatal mortality in captive axis deer. To the best of our knowledge, this is the first report of clinical disease associated to N. caninum infection in axis deer and of isolation of the parasite from this wild ruminant species.  相似文献   

Effect of olive-derived antioxidants (3,4-dihydroxyphenylethanol and 3,4 dihydroxyphenylglycol) on sperm motility and fertility in liquid ram sperm stored at 15°C or 5°C     

Ander Arando  Juan Vicente Delgado  Alejandra Bermúdez-Oria  José Manuel León  África Fernández-Prior  Sergio Nogales  Carlos C. Pérez-Marín 《Reproduction in domestic animals》2020,55(3):325-332

The aim of the present study was to evaluate the effect of the addition of two olive oil-derived antioxidants, hydroxytyrosol (3,4-dihydroxyphenylethanol, HT) and 3,4-dihydroxyphenylglycol (DHPG), on ovine semen during liquid storage at 5°C and 15°C. Semen was collected, pooled, diluted and then divided into aliquots supplemented with different concentrations (5 μg/ml, 10 μg/ml, 50 μg/ml and 100 μg/ml) of HT, DHPG and a mixture (MIX) of both antioxidants. Sperm motility characteristics were assessed in the different samples at 0, 6, 24, 48, 72 and 96 hr after cooling, and a fertility trial was also conducted. The results showed that the antioxidant addition did not significantly improve total and progressive motility in ovine cooled sperm maintained at 15° or 5°C. However, in samples stored at 5°C, LIN (48, 72, 96 hr), STR (0 hr) and WOB (0, 48, 72, 96 hr) values significantly decreased in comparison with control treatment when high antioxidant concentrations were added (MIX100 or HT100). When samples were maintained at 15°C, MIX50 showed significantly higher VCL values than the control treatment after 6 hr cooling, and MIX100 showed significantly lower VCL values at 96 hr after cooling. According to the artificial insemination trial, no significant differences were observed when antioxidants were added. In conclusion, the use of HT and DHPG showed small impact in sperm motility and fertility was not affected (nor detrimentally nor positively) when insemination was carried out using antioxidant-supplemented liquid sperm.  相似文献   

Distribution of whale shark (Rhincodon typus) off northern Peru based on habitat suitability     

Adriana Gonzalez‐Pestana  Rossana Maguio  Alejandra Mendoza  Shaleyla Kelez  Dení Ramírez‐Macías 《水产资源保护:海洋与淡水生态系统》2020,30(7):1325-1336

1. In the south‐eastern Pacific Ocean, few studies of whale sharks (Rhincodon typus) exist. In Peru, the northern coast has been identified as the area with the highest presence of whale sharks, yet their ecology in this area is poorly defined.
2. This study predicts the spatial distribution of whale sharks off coastal northern Peru (03°00′S–04°30′S) during La Niña and El Niño seasonal conditions, utilizing maximum entropy modelling. Between 2009 and 2018 (except for 2011), 347 whale sharks were geo‐referenced in northern Peru with greatest data recordings in the austral summer and spring during La Niña events.
3. Depth was the most important predictive variable for spatial distribution of whale sharks, followed by chlorophyll‐a. Sharks were predicted in shallower coastal waters in which chlorophyll‐a values are higher.
4. Habitat suitability was higher in the northern coastal part of the study area. Spring presents the most suitable environmental conditions for whale sharks, both during La Niña and El Niño conditions. The probability of whale shark presence in the north of Peru increases at higher chlorophyll‐a and sea surface temperature values. Therefore, whale sharks appear to aggregate seasonally in northern Peru, potentially exploiting rich foraging grounds.
5. In these areas of high suitability, whale sharks are susceptible to fisheries, bycatch, ship collisions, unmanaged tourism, and pollution; thus, management actions should focus in these areas.
6. This study represents a first step to understand the distribution and habitat suitability of whale shark in Peruvian waters. Further studies should identify suitable habitat for whale sharks in offshore areas. Also, these should focus on the connectivity of these aggregations with other localities in the south‐eastern Pacific in order to contribute to regional strategies for the conservation of this iconic species in this particular region.

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