Triploid Induction in the Yellowtail Tetra,Astyanax altiparanae,Using Temperature Shock: Tools for Conservation and Aquaculture

Triploidization is an interesting tool to produce sterile fish. In the yellowtail tetra, Astyanax altiparanae, this can be applied for aquaculture and surrogate technologies. In this study, we compared the efficacy of cold (2 C) or heat shock (38 C, 40 C, and 42 C) on triploid induction in the yellowtail tetra. The eggs were treated with cold or heat shock, 2 min postfertilization (30 min in cold shock or 2 min in heat shock). Intact embryos served as the control group. Ploidy status was confirmed by karyotyping, flow cytometry, and nuclear diameter of erythrocytes. The hatching rate decreased after cold shock (12.69 ± 15.76%) and heat shock at 42 C (0.35 ± 0.69%) in comparison with the control group (63.19 ± 16.82%). At 38 C and 40 C, hatching rates (61.29 ± 17.73% and 61.75 ± 22.1%, respectively) were not decreased. Only one triploid arose at 38 C (1/80). At 40 C, a high number of triploids arose (72/78). At 42 C, very few embryos developed into the hatching stage. A large number of haploid individuals arose after cold shock (61/75), with only one triploid. Our results indicate that heat shocking of embryos at 40 C is optimum for triploid production in the yellowtail tetra.     

Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.     

Study of Resistance to Orobanche ramosa in Host (Oilseed Rape and Carrot) and Non-host (Maize) Plants

Orobanche ramosa is a parasitic Angiosperm responsible for severe yield losses in several economical crops. It is a serious threat in oilseed rape in France and Morocco and is appearing in carrot crops in Morocco. In this study, several varieties of oilseed rape and carrot were screened in order to identify resistant cultivars and to characterize the resistance mechanisms involved. All the 15 oilseed rape varieties tested were susceptible. In carrot, the varieties 'Colmar à coeur rouge' and 'Nantaise demi-longue' were susceptible, whereas 'Palaiseau' and 'Buror' were resistant. In the susceptible 'Colmar à coeur rouge' carrot no defence reactions were found and the development of the parasite inhibited carrot tap root formation. In the resistant carrot varieties, the parasite germinated, became attached to the host root but became necrotic before emergence. In 'Buror' carrot, formation of a mechanical barrier was associated with the restriction to the cortex of the parasite. In maize cv. 'Vigni', a non-host of O. ramosa, thickening of xylem vessels, cell divisions in the central cylinder and formation of an encapsulation layer were observed in association with restricted development of Orobanche haustoria.     

Dispersal of conidia of Fusicladium eriobotryae and spatial patterns of scab in loquat orchards in Spain

Dispersal of conidia of Fusicladium eriobotryae, the causal agent of loquat scab, was investigated in two loquat orchards in Spain from 2010 to 2012. A volumetric spore sampler, horizontally and vertically orientated microscope slides, and rain collectors were placed in loquat fields to trap conidia of F. eriobotryae. No conidia were collected in the volumetric sampler, and highly variable numbers of conidia were collected in the rain gatherers. Large numbers of conidia were collected by microscope slides, particularly by those held in a horizontal orientation compared with those held in a vertical orientation. Approximately 90 % of the F. eriobotryae conidia were collected during rainy periods. Based on ROC and Bayesian analysis, using ≥ 0.2 mm rainfall as a cut-off value resulted in a high probability of correctly predicting actual conidial dispersal, and had a low probability of failing to predict actual conidial dispersal. Based on the index of dispersion and the binary power law, the incidence of loquat scab on fruit was highly aggregated in space between and within trees, and aggregation was influenced by disease incidence. Our results demonstrate, for the first time, that F. eriobotryae is dispersed mainly in rain splash. The results will be integrated into a mechanistic, weather-driven, disease prediction model that should help growers to minimize fungicide application for the management of loquat scab.     

SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis

Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.     

Uterine neoplasia in 13 cats.

Thirteen uterine tumors were diagnosed in 13 cats and accounted for 0.29% of all feline neoplasms received during a 9.6-year period. Age at diagnosis ranged from 3 to 16 years; median 9 years. Six were Domestic Shorthair cats, and 7 were purebred cats of 5 different breeds. Eight adenocarcinomas and 1 mixed Müllerian tumor (adenosarcoma) comprised the endometrial tumors. Myometrial tumors included 3 leiomyomas and 1 leiomyosarcoma. One of the adenocarcinomas developed in the uterine stump of an ovariohysterectomized cat; the other cats were sexually intact. Concurrent mammary adenocarcinoma was diagnosed in 1 cat with uterine adenocarcinoma and in another with uterine leiomyoma. Tumors were discovered during elective ovariohysterectomy in 2 cats, but at least 3 others had experienced reproductive problems (infertility or pyometra). Five cats presented for abdominal or pelvic masses. Endometrial adenocarcinomas were positive immunohistochemically for cytokeratins and negative for smooth muscle actin (SMA): 1 of 6 cats was positive for vimentin and 4 of 8 were positive for estrogen receptor-alpha (ER alpha). Adenosarcoma stromal cells were positive for vimentin and ER alpha but negative for cytokeratins and SMA. Smooth muscle tumors were positive for vimentin and SMA and negative for cytokeratins. Leiomyomas, but not the leiomyosarcomas, were positive for ER alpha. Adenocarcinomas in 4 cats had metastasized by the time of ovariohysterectomy. Two other cats were euthanized 5 months after ovariohysterectomy; at least one of these cats had developed an abdominal mass that was not examined histologically. Only 2 cats with endometrial adenocarcinoma had disease-free intervals longer than 5 months after surgery. Metastasis was not detected in any mesenchymal tumor; however, these cats were either euthanized on discovery of the tumor or the tumor was first detected at necropsy.     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

Virulence factors in Escherichia coli isolated from the blood of bacteremic neonatal calves

Twenty-five Escherichia coli isolates from the blood of critically ill bacteremic calves sampled in two separate studies on a calf-rearing farm housing over 15,000 calves, in the San Joaquin Valley, California were studied.Isolates were characterized for O serogroups and for pathotypes as determined by the presence of specific virulence factors including heat-labile enterotoxin (LT), heat-stable enterotoxins a and b (STa, STb), verotoxins 1 and 2 (VT1, VT2), cytotoxic necrotizing factor (CNF), aerobactin, intimin Eae and P, F17 and CS31A fimbrial adhesins, and resistance to bactericidal effects of serum.These isolates constituted a heterogeneous group. However, isolates were mostly aerobactin positive and often resistant to the bactericidal effects of serum. Isolates of pathotypes O78 (n=6), O119:CS31a (n=3), and P positive but O non-typeable (n=3) were associated with a high mortality rate. The remaining isolates belonged to diverse pathotypes, often possessing the adhesins P, F17, CS31A and Eae but belonging to O serogroups other than O78 and O119, and were less frequently associated with mortality.Although no virulence factor common to all isolates was identified, the capacity to use iron by the presence of aerobactin which is important to the capture of iron was a predominant factor. Moreover, certain pathotypes appear to be associated with primary colisepticemia whereas other pathotypes may cause a bacteremia without necessarily leading to septicemia.     

Open reduction of sacral fractures using transarticular implants at the articular facets of L7-S1: 8 consecutive canine patients (1995-1999)

OBJECTIVE: To describe and evaluate facet stabilization as an aid for open reduction and fixation of sacral fractures. STUDY DESIGN: A clinical case series. STUDY POPULATION: Eight consecutive client-owned dogs with sacral fractures. METHODS: A combination of a standard lateral approach to the ilium and a dorsal approach to the articular facets of L7-S1 was performed. The articular facets were reduced and stabilized with a Kirschner wire or a combination of a Kirschner wire and a cortical screw. Primary stabilization of the sacrum was then accomplished using a cortical screw inserted from the lateral aspect of the ilium, through the sacral fracture into the body of the sacrum. Accuracy of reduction, position of the implants, percent screw depth/sacral width, pelvic canal diameter ratio, and complications were assessed using radiographs and physical examination. RESULTS: Mean sacral fracture reduction was 91% immediately after surgery, and was 87% when measured at follow-up examinations. Mean screw depth/sacral width was 66%. Mean pelvic canal diameter ratio was 1.12 postoperatively and 1.03 at the 4- to 6-week reevaluation. Six of the 8 dogs were using the leg well 10 to 12 days after surgery; none had clinically detectable neurologic deficits. The long-term (mean, 2.7 years) outcome for these dogs was good (normal to grade I lameness). CONCLUSION: Fixation of the L7-S1 facet joints facilitates repair of sacral fractures.     

Does reverse growth dominance develop in old plantations of Eucalyptus saligna?

Patterns of stand growth and dominance among individual trees change through stand development. We tested a prediction that large, old plantations of Eucalyptus saligna (Sm.) in Hawaii and Brazil would show a pattern of “reverse” growth dominance, with the largest trees contributing more to stand biomass than to current stand increment. This pattern might develop if the largest trees accounted for a large portion of the total resource use of the stand, but were less efficient at utilizing resources to produce stemwood. The 70-year-old plantation in Hawaii had a stem mass of 610 Mg/ha, compared with 325 Mg/ha in the 66-year-old Brazil plantation. Cumulative frequency plots for both sites showed that dominant trees comprised a very large proportion of total stand mass; the largest 20% of the trees accounted for 50% and 30% of the total stand mass in Hawaii and Brazil, respectively. Contrary to expectations, the largest trees continued to produce a disproportionately large amount of total stand growth, with the largest 20% of trees accounting for 60% (Hawaii) and 40% (Brazil) of stand growth. The frequency distributions resulted in positive growth dominance coefficients (similar to Gini coefficients) of 0.14 for Hawaii and 0.09 for Brazil, rather than negative values that would indicate that reverse growth dominance. The failure of these monoculture Eucalyptus stands to develop reverse growth dominance contrasts starkly with a variety of studies with pine species, which tend to show neutral-to-reverse growth dominance. Indeed, continued dominance of large trees suggests the trees might respond well to silvicultural treatments such as thinning or fertilization. Future work that contrasts patterns of growth dominance among species will be important as a foundation for developing generalizations about trends in growth dominance with stand development.