Anatomicohistological Characteristics of the Ovary of the Coypu (Myocastor coypus)

Using histological, histochemical and macroscopic and microscopic measurement techniques, the macroscopic and microscopic structures of coypu ovaries were studied in sexually mature virgin females. The mature ovaries of the coypu were ovoid or elongated bodies, not encapsulated, covered by a single layer of epithelium. They had a parenchyma formed by follicles at different stages of evolution and true and accessory corpora lutea. The interstitial tissue was a prominent and permanent structure in the ovaries. Some ovaries contained a few rete ovarii in the hilus.     

Serological profile of Toxoplasma gondii and Neospora caninum infection in commercial sheep from São Paulo State, Brazil

Neosporosis and toxoplasmosis are two important infections in young and adult sheep, leading to low production and abortion. This study aimed to determine the frequency of antibodies to Toxoplasma gondii and Neospora caninum in sheep from the eastern region of S?o Paulo State, Brazil. Serum samples (382) were collected from the sheep and assayed for T. gondii through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT), and for N. caninum antibodies, through IFAT, with cut-off titers equal to 16 (T. gondii) and 25 (N. caninum). All frozen samples were sent to the Center for Zoonoses Research (NUPEZO), Department of Veterinary Hygiene and Public Health (DHSVP), FMVZ, UNESP, for serological tests. A total of 71/382 (18.6%) samples reacted to T. gondii, especially at titers 16 (28; 39.4%), 64 (15; 21.1%), 256 (21; 29.6%) and 1024 (6; 8.5%) by MAT, and 16 (34; 47.9%), 64 (18; 25.4%), 256 (14; 19.7%) and 1024 (5; 7%) by IFAT. As regards N. caninum, 49/382 (12.8%) samples reacted at titers 25 (17; 34.7%), 50 (11; 22.5%), 100 (11; 22.5%), and ≥ 200 (10; 20.4%). These animals presented infection but no clinical signs. Six and ten animals had high titers for toxoplasmosis and neosporosis. No significant association was observed between antibodies for both parasites (P=0.535) according to Fisher's exact test, and no correlation was found between T. gondii (MAT) and N. caninum antibody titers (r=-0.0068; P=0.895), T. gondii (IFAT) and N. caninum antibody titers (r=-0.0025; P=0.961). Thus, T. gondii and N. caninum infections were observed in farms located in S?o Paulo State, where sheep play an important economical role for the national and regional business.     

Meta-analysis of the prevalence and risk factors associated with bovine neosporosis

Tropical Animal Health and Production - Neospora caninum is one of the main infectious agents that cause reproductive disorders in cattle. However, knowledge about the prevalence and causal factors...     

Tissue‐print and squash real‐time PCR for direct detection of ‘Candidatus Liberibacter’ species in citrus plants and psyllid vectors

Huanglongbing (HLB) disease is seriously threatening and/or damaging the citrus industry worldwide. Accurate detection of the three species associated with HLB disease, ‘Candidatus Liberibacter asiaticus’, ‘Candidatus Liberibacter africanus’ and ‘Candidatus Liberibacter americanus’, is essential for the preventive control of the disease. Real‐time PCR is a useful tool for bacterial detection. However, nucleic acid purification steps limit the number of samples that can be processed by PCR. Universal detection of ‘Ca. Liberibacter’ species was achieved by direct tissue‐printing and spotting of plant leaf petiole extracts or squashing of individual psyllids onto paper or nylon membranes. Primers were designed and used with TaqMan chemistry for accurate detection of the bacterium in immobilized targets (prints of 10 overlapping leaf pedicels per tree, or squashed single vectors), by extraction with water and direct use for real‐time PCR. This simplified method was validated and could detect HLB‐liberibacters in 100% of leaves with symptoms and 59% of symptomless leaves collected from HLB‐infected trees. The use of direct assays as template showed good agreement with use of purified DNA (κ = 0·76 ± 0·052). The squash assay allowed detection of the bacterium in 40% of mature Diaphorina citri that fed on leaves of HLB‐infected trees with or without symptoms. A commercial ready‐made kit based on this technology showed 96% accuracy in intra‐laboratory performance studies. The simplified direct methods of sample preparation presented herein can be effectively adopted for use in rapid screening of HLB agents in extensive surveys, certification schemes or for epidemiological and research studies.     

Comparative study of genetic diversity of Clavibacter michiganensis subsp. michiganensis isolates from the Canary Islands by RAPD-PCR, BOX-PCR and AFLP

Molecular characterization of seedborne pathogens is an important issue when discerning their origin and tracking the spread of a disease. In the Canary Islands (Spain), Clavibacter michiganensis subsp. michiganensis (Cmm) was first detected in 2002, causing severe losses in many tomato-growing areas. Fifty four strains of this bacterium isolated from 2002 to 2007 and 19 strains from different countries were characterized for genetic diversity. RAPD-PCR, BOX-PCR and AFLP provided differentiation among Cmm strains whereas no differences were observed with ERIC-PCR, REP-PCR and 16S-23S ITS PCR-RFLP. RAPD-PCR and BOX-PCR revealed high homogeneity among the Canary Island strains (>80 and >75% of similarity, respectively) which could not be grouped based on tomato cultivar, location or year of isolation. By contrast, strains of Cmm from other countries displayed high diversity, providing several clusters, most of which were composed of a single strain. Similarly, AFLP analysis of 29 selected strains of Cmm gave the same profile for the Canarian ones (>90% of similarity) whereas high polymorphism was obtained with strains from different countries. Moreover, two strains, one from the USA and another from Spain, were related to the Canarian strains, according to RAPD-PCR (>60% of similarity), BOX-PCR (>75%) and AFLP analysis (>90%), suggesting a common origin. The circumstances under which the Cmm outbreaks occurred in the Canary Islands and the high homogeneity observed among the Canarian strains would suggest that the bacterium was introduced into the region from only one origin.     

Pewenomyces kutranfy gen. nov. et sp. nov. causal agent of an important canker disease on Araucaria araucana in Chile

Araucaria araucana, (commonly referred to as araucaria, pewen, or monkey puzzle tree) is an ancient conifer endemic to the Chilean and Argentinian mountain ranges where it has a sacred relevance to indigenous communities. During 2015, a serious disease was noticed on trees of all ages in most of the natural distribution of this iconic tree. Four areas were surveyed, and the most important symptoms of the disease were cankers on branches and stems resulting in copious resin exudation. Trees were monitored for a period of two years and isolations were made from the cankers. Field observations showed that the disease typically begins on the leaves or at the leaf bases and progresses downwards to initiate cankers that can girdle branches or stems within a two-year period. Black ascomata, resembling those of Caliciopsis species previously described from A. araucana, were consistently found developing in the cankers from which isolations were made. Phylogenetic analyses of the ITS, nucSSU, and nucLSU gene regions showed that the fungus resides in the Coryneliaceae but is distinct from other genera in that family. The morphological characteristics and phylogenetic position of the fungus show that it represents a new genus and species, described here as Pewenomyces kutranfy gen. nov. et sp. nov. Pathogenicity trials on trees under field conditions confirmed that this newly described fungus is able to cause cankers on A. araucana similar to those found under natural conditions.     

Detection of Bemisia tabaci Mediterranean cryptic species on soybean in São Paulo and Paraná States (Brazil) and interaction of cowpea mild mottle virus with whiteflies

Since the detection of Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) species in Brazil in the early 1990s, this whitefly has been the predominant species and the main viral vector in the country. In this work, whiteflies were collected from commercial soybean fields near and far from greenhouses where Mediterranean (MED) species had previously been detected infesting vegetable crops. Results indicated that MEAM1 was the predominant whitefly species in soybean in most sampling sites, while MED was found colonizing soybean plants in open field conditions alone and/or with MEAM1 in several places. Among the tested insects, MED species was mostly detected harbouring the facultative endosymbiont Hamiltonella. We also detected cowpea mild mottle virus (CPMMV) infecting soybean plants. Transmission assays demonstrated that MED species was more efficient transmitting CPMMV from beans to beans and from soybean to soybean plants than MEAM1, while MEAM1 was more efficient in transmitting the virus from soybean to bean than MED. Performance assays indicated that adult emergence of both species on CPMMV-infected soybean plants was higher when compared with the emergence on healthy plants. Moreover, nonviruliferous MED and MEAM1 adults preferred to settle more often on healthy plants, while viruliferous adults settled more often on CPMMV-infected soybean plants. As MED has already been found in soybean open fields in São Paulo and Paraná States, Brazil, and it is a good vector of CPMMV, we anticipate problems related to whitefly management and to increase in the incidence of the virus in soybean.