REP-PCR Analysis of Erwinia Genus Bacteria – Infectious Agents of Apple Trees Diseases in Ukraine

2019 ◽  
Vol 81 (6) ◽  
pp. 45-57
Author(s):  
L.A. Dankevych ◽  
◽  
F.V. Muchnyk ◽  
V.P. Patyka ◽  
◽  
...  
Keyword(s):  
Pcr Analysis ◽  
Infectious Agents ◽  
Apple Trees

scholarly journals Incidence and Molecular Identification of Apple Necrotic Mosaic Virus (ApNMV) in Southwest China

Plants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 415
Author(s):  
Wensen Shi ◽  
Rundong Yao ◽  
Runze Sunwu ◽  
Kui Huang ◽  
Zhibin Liu ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Southwest China ◽  
Great Influence ◽  
Mosaic Disease ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Apple Trees ◽  
Apple Leaves ◽  
Prunus Necrotic Ringspot Virus ◽  
Apple Production

Apple mosaic disease has a great influence on apple production. In this study, an investigation into the incidence of apple mosaic disease in southwest China was performed, and the pathogen associated with the disease was detected. The results show that 2869 apple trees with mosaic disease were found in the Sichuan, Yunnan, and Guizhou Provinces, with an average incidence of 9.6%. Although apple mosaic virus (ApMV) is widespread in apples worldwide, the diseased samples were negative when tested for ApMV. However, a novel ilarvirus (apple necrotic mosaic virus, ApNMV) was identified in mosaic apple leaves which tested negative for ApMV. RT-PCR analysis indicated that ApNMV was detected in 322 out of 357 samples with mosaic symptoms. Phylogenetic analysis of coat protein (CP) sequences of ApNMV isolates suggested that, compared with ApMV, ApNMV was closer to prunus necrotic ringspot virus (PNRSV). The CP sequences of the isolates showed the diversity of ApNMV, which may enable the virus to adapt to the changeable environments. In addition, the pathology of mosaic disease was observed by microscope, and the result showed that the arrangement of the tissue and the shape of the cell, including the organelle, were seriously destroyed or drastically changed.

scholarly journals Post-mortem hemoparasite detection in free-living Brazilian brown brocket deer (Mazama gouazoubira, Fischer 1814)

2014 ◽  
Vol 23 (2) ◽  
pp. 206-215 ◽  
Author(s):  
Júlia Angélica Gonçalves da Silveira ◽  
Élida Mara Leite Rabelo ◽  
Paula Cristina Senra Lima ◽  
Bárbara Neves Chaves ◽  
Múcio Flávio Barbosa Ribeiro
Keyword(s):  
Nested Pcr ◽  
Pcr Analysis ◽  
Post Mortem ◽  
Infectious Agents ◽  
Free Living ◽  
Molecular Assays ◽  
Minas Gerais State ◽  
Mazama Gouazoubira ◽  
Nested Pcr Assays ◽  
Brocket Deer

Tick-borne infections can result in serious health problems for wild ruminants, and some of these infectious agents can be considered zoonosis. The aim of the present study was the post-mortem detection of hemoparasites in free-living Mazama gouazoubira from Minas Gerais state, Brazil. The deer samples consisted of free-living M. gouazoubira (n = 9) individuals that died after capture. Necropsy examinations of the carcasses were performed to search for macroscopic alterations. Organ samples were collected for subsequent imprint slides, and nested PCR assays were performed to detect hemoparasite species. Imprint slide assays from four deer showed erythrocytes infected with Piroplasmida small trophozoites, and A. marginale corpuscles were observed in erythrocytes from two animals. A. marginale and trophozoite co-infections occurred in two deer. A nested PCR analysis of the organs showed that six of the nine samples were positive for Theileria sp., five were positive for A. phagocytophilum and three were positive for A. marginale, with co-infection occurring in four deer. The results of the present study demonstrate that post-mortemdiagnostics using imprint slides and molecular assays are an effective method for detecting hemoparasites in organs.

scholarly journals 138EARLY EMBRYO MORTALITY IN BUFFALO (BUBALUS BUBALIS) DURING THE SPRING PERIOD

2004 ◽  
Vol 16 (2) ◽  
pp. 191
Author(s):  
G. Neglia ◽  
C. Grassi ◽  
A. Prandi ◽  
G. Galiero ◽  
E. De Carlo ◽  
...  
Keyword(s):  
Bubalus Bubalis ◽  
Early Embryo ◽  
Pcr Analysis ◽  
Blood Levels ◽  
Infectious Agents ◽  
National Meeting ◽  
Embryo Mortality ◽  
Pregnancy Diagnosis ◽  
Spring Period ◽  
Group B

Embryo development is linked to several factors, such as intrinsic defects within the embryo, an inadequate maternal environment or failure of maternal response to embryonic signals. Reproductive seasonality in buffalo species (Bubalus bubalis) is responsible for the lower reliability of the pregnancy diagnosis by progesterone assay in spring, probably due to the high incidence of early embryo mortality (EEM) for hormonal reasons (Campanile et al., 1989; Proc. 2th National Meeting ‘Studio dell’efficienza riproduttiva degli animali di interesse zootecnico’: 51–57). The aim of the study was to identify the causes of EEM in buffaloes inseminated during the spring period. Italian Mediterranean Buffalo cows (n=209) in a good state of health were synchronized using the Ovsynch-TAI Program (Pursley et al., 1995 Theriogenology 44, 915–923) and inseminated 16 and 40 hours after the second administration of GnRH. Artificial inseminations were performed by the same technician. Ten to 20 days after the first insemination two blood samples were collected from each animal for assessing P4 levels by RIA. Days 24 and 40 after the first insemination, pregnancy diagnosis was assessed by rectal ultrasonography (Aloka SSD-500) with a 5MHz sector scanner. In animals with EEM a uterine flushing was carried out and recovered fluid was analyzed for the principal infectious agents. Microbiological analyses were carried out according to Quinn et al. (1994 Clinical Veterinary Microbiology, Wolfe Publishing, Mosby, Europe) and a PCR analysis was performed only for Bubaline Herpes Virus (BuHV1), Toxoplasma and Neospora. Statistical analysis was assessed by Student’s t-test. The incidence of cyclic buffaloes after synchronization treatment (94%) and pregnancy rate at 40 days (34.4%) were similar to those reported in a previous study (Neglia et al., 2003 Theriogenology 60, 125–133). EEM between 24 and 40 days was 45% and only in 9 buffaloes (15.3%), these results correlated with the presence of infectious agents (Streptococcus spp. group B and G, Bacillus licheniformis, Proteus mirabilis, Staphylococcus spp.). P4 levels on Day 10 were higher (P<0.05) in pregnant (P) buffaloes than in those with EEM. On Day 20, P4 levels were higher (P<0.01) in P buffaloes than in those with EEM and those not pregnant. It is hypothesized that the reduced activity of hypophysis, correlated with an increase in hours of light, may be responsible for decreased P4 concentrations and EEM in buffalo. Table 1 P4 blood levels in pregnant (P) and non pregnant (NP) buffaloes and in buffaloes with EEM 10 and 20 days after insemination

scholarly journals A Nested PCR Assay for Detecting Valsa mali var. mali in Different Tissues of Apple Trees

Plant Disease ◽  
2012 ◽  
Vol 96 (11) ◽  
pp. 1645-1652 ◽  
Author(s):  
Rui Zang ◽  
Zhiyuan Yin ◽  
Xiwang Ke ◽  
Xiaojie Wang ◽  
Zhengli Li ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Detection Rate ◽  
Pcr Analysis ◽  
Pcr Assay ◽  
Apple Trees ◽  
Specific Primers ◽  
Fungal Isolation ◽  
Species Specific ◽  
Valsa Mali ◽  
Different Tissues

A nested polymerase chain reaction (PCR) assay for detecting Valsa mali var. mali, the causal agent of apple tree Valsa canker, was developed. One pair of genus-specific primers was designed based on the ribosomal DNA internal transcribed spacer conservative sequence of the Valsa genus and one pair of species-specific primers was designed based on the specific sequence of V. mali var. mali. The specificity of the genus-specific and species-specific primers was evaluated against 10 V. mali var. mali isolates, 10 V. mali var. pyri isolates, 4 isolates from closely related Valsa spp., and 8 isolates from fungal species that are commonly isolated from naturally infected apple bark tissue. A distinct band of 348 bp in length was detected in all V. mali var. mali isolates but not in other tested species and the V. mali var. pyri variety. The sensitivity of this assay was evaluated by serial dilutions of DNA extracted from V. mali var. mali pure cultures and apple bark tissues with or without visible symptoms. The results showed that the assay was able to detect as little as 100 fg of DNA in mycelial samples and apple bark tissues with visible symptoms, whereas the lowest detectable concentration was 10 pg of DNA in symptomless apple bark tissues. The efficiency of the nested PCR assay was compared with that of fungal isolation assays. All symptomless and symptomatic samples from which the pathogen was successfully isolated yielded a PCR product of the expected size. The detection rate of nested PCR for symptomless samples was 64.7%, which was much higher than the detection rate of 20.6% by fungal isolation. The PCR analysis of different symptomless tissues showed that the incidence of V. mali var. mali was different in different tissues of apple trees. The average incidence of V. mali var. mali was 89% in terminal buds, 71% in internodes, and 48% in bud scale scars. Moreover, the incidence of V. mali var. mali in nonsymptomatic tissues was higher in orchards where more trees were infected. Taken together, the assay developed in this study can be used for rapid and reliable detection of V. mali var. mali in tissues of apple trees with or without symptoms and also for monitoring the presence of the pathogen at an early stage of disease development.

Pathology of Bone Cells in Pigs with Experimental Turbinate Osteoporosis (Atrophic Rhinitis)

1971 ◽  
Vol 29 ◽  
pp. 304-305
Author(s):  
A. W. Fetter ◽  
C. C. Capen
Keyword(s):  
Mucous Membrane ◽  
Bone Cells ◽  
Atrophic Rhinitis ◽  
Infectious Agents ◽  
Metabolic Disturbances ◽  
Progressive Reduction ◽  
Naturally Occurring ◽  
Fundamental Cause ◽  
Osteocytic Osteolysis ◽  
Uncertain Etiology

Atrophic rhinitis in swine is a disease of uncertain etiology in which infectious agents, hereditary predisposition, and metabolic disturbances have been reported to be of primary etiologic importance. It shares many similarities, both clinically and pathologically, with ozena in man. The disease is characterized by deformity and reduction in volume of the nasal turbinates. The fundamental cause for the localized lesion of bone in the nasal turbinates has not been established. Reduced osteogenesis, increased resorption related to inflammation of the nasal mucous membrane, and excessive resorption due to osteocytic osteolysis stimulated by hyperparathyroidism have been suggested as possible pathogenetic mechanisms.The objectives of this investigation were to evaluate ultrastructurally bone cells in the nasal turbinates of pigs with experimentally induced atrophic rhinitis, and to compare these findings to those in control pigs of the same age and pigs with the naturally occurring disease, in order to define the fundamental lesion responsible for the progressive reduction in volume of the osseous core.

Characterization of Partial Gene Deletions in Type III von Willebrand Disease with Alloantibody Inhibitors

1994 ◽  
Vol 72 (02) ◽  
pp. 180-185 ◽  
Author(s):  
David J Mancuso ◽  
Elodee A Tuley ◽  
Ricardo Castillo ◽  
Norma de Bosch ◽  
Pler M Mannucci ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Pcr Analysis ◽  
Gene Deletions ◽  
Factor Gene ◽  
Type Iii ◽  
Large Deletions ◽  
Von Willebrand ◽  
Willebrand Factor

Summaryvon Willebrand factor gene deletions were characterized in four patients with severe type III von Willebrand disease and alloantibodies to von Willebrand factor. A PCR-based strategy was used to characterize the boundaries of the deletions. Identical 30 kb von Willebrand factor gene deletions which include exons 33 through 38 were identified in two siblings of one family by this method. A small 5 base pair insertion (CCTGG) was sequenced at the deletion breakpoint. PCR analysis was used to detect the deletion in three generations of the family, including two family members who are heterozygous for the deletion. In a second family, two type III vWD patients, who are distant cousins, share an -56 kb deletion of exons 22 through 43. The identification and characterization of large vWF gene deletions in these type III vWD patients provides further support for the association between large deletions in both von Willebrand factor alleles and the development of inhibitory alloantibodies.

Exposure of the Gulf of St. Lawrence grey seal Halichoerus grypus population to potentially zoonotic infectious agents

2020 ◽  
Vol 142 ◽  
pp. 105-118
Author(s):  
CC Sauvé ◽  
A Hernández-Ortiz ◽  
E Jenkins ◽  
F Mavrot ◽  
A Schneider ◽  
...  
Keyword(s):  
Meat Products ◽  
Halichoerus Grypus ◽  
Human Consumption ◽  
Infectious Agents ◽  
Erysipelothrix Rhusiopathiae ◽  
Public Health Significance ◽  
Domestic Livestock ◽  
Health Significance ◽  
Commercial Source ◽  
Polymerase Chain

The population of grey seals Halichoerus grypus in Canadian waters is currently used as a commercial source of meat for human consumption. As with domestic livestock, it is important to understand the occurrence in these seals of infectious agents that may be of public health significance and thus ensure appropriate measures are in place to avoid zoonotic transmission. This study examined the prevalence of antibodies against Brucella spp., Erysipelothrix rhusiopathiae, 6 serovars of Leptospira interrogans, and Toxoplasma gondii in 59 grey seals and determined by polymerase chain reaction (PCR) the presence of these potentially zoonotic agents in specific organs and tissues of seropositive animals. The presence of encysted Trichinella spp. larvae was also investigated by digestion of tongue, diaphragm and other muscle samples, but none were detected. Seroprevalence against Brucella spp. and E. rhusiopathiae was low (5 and 3%, respectively). All 59 seals tested had antibodies against L. interrogans, but no carrier of this bacterium was detected by PCR. Seroprevalence against T. gondii was 53%, and DNA of this protozoan was detected by PCR in 11/30 (37%) seropositive animals. Standard sanitary measures mandatory for commercialization of meat products for human consumption should greatly reduce the potential for exposure to these infectious agents. However, special consideration should be given to freezing seal meat for at least 3 d to ensure destruction of tissue cysts of T. gondii.

Multiplex PCR Analysis of Ultraviolet A and B Induced Delayed and Early Mutations in V79 Chinese Hamster Cells

2004 ◽  
Author(s):  
Jostein Dahle ◽  
Paul Noordhuis ◽  
Trond Stokke ◽  
Debbie Hege Svendsrud ◽  
Egil Kvam
Keyword(s):  
Multiplex Pcr ◽  
Chinese Hamster ◽  
Pcr Analysis ◽  
Ultraviolet A ◽  
Chinese Hamster Cells
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