Evaluation of Bacillus spp. isolates as potential biocontrol agents against charcoal rot caused by Macrophomina phaseolina on common bean

The fungus Macrophomina phaseolina, the causal agent of charcoal rot of common beans, damages the roots, stems, and leaves of seedlings and plants and forms resistant structures, so that chemicals are not sufficient for disease control. Integrated management systems associated with the use of biological control techniques are a sustainable alternative. Here we collected 37 native bacterial isolates from the common bean rhizosphere and screened them for antagonistic activity against M. phaseolina. Four isolates (BA97, BN17, BN20, and BR20) identified as Bacillus spp. showed antagonism in vitro against M. phaseolina, inhibiting its growth by 62.5–85%. In an in planta antagonistic assay, isolate BN20 reduced disease severity the most. Isolates BA97, BN17, BN20, and BR20 produced volatile compounds as a mechanism of antagonism. They also produced indole acetic acid in vitro (1.98–3.87 μg/ml). These results suggest that seed bacterization with the rhizobacterial isolates for field planting may be an effective means to reduce crop damage by M. phaseolina.

     

Single layer centrifugation-selected boar spermatozoa are capable of fertilization in vitro

Background

Good quality spermatozoa are important to achieve fertilization, viable embryos and offspring. Single Layer Centrifugation (SLC) through a colloid (Androcoll-P) selects good quality spermatozoa. However, it has not been established previously whether porcine spermatozoa selected by this method maintain their fertility.

Methods

The semen was prepared either by SLC or by standard centrifugation (control) and used for in vitro fertilization (IVF) at oocyte:spermatozoa ratios of 1:50; 1:100 and 1:300 (or 4 x 10³, 8 x 10³ and 24 x 10³ spermatozoa/ml) to evaluate their subsequent ability to generate blastocysts. In addition, sperm motility was assessed by computer assisted sperm motility analysis.

Results

Total and progressive motility were significantly higher in sperm samples prepared by SLC compared to uncentrifuged samples. Sperm binding ability, polyspermy, cleavage and blastocyst rates were affected by the oocyte:sperm ratio, but not by sperm treatment.

Conclusion

The use of SLC does not adversely affect the in vitro fertilizing and embryo-generating ability of the selected spermatozoa compared to their unselected counterparts, but further modifications in the IVF conditions would be needed to improve the monospermy in IVF systems. Since SLC did not appear to have a negative effect on sperm fertilizing ability, and may in fact select for spermatozoa with a greater potential for fertilization, an in vivo trial to determine the usefulness of this sperm preparation technique prior to artificial insemination is warranted.     

Diagnostic value of the use of lateral and occlusal radiographic views in comparison with periodontal probing for the assessment of periodontal attachment of the canine teeth in dogs

OBJECTIVE: To determine the diagnostic value of 2 intraoral bisecting angle radiographic views in comparison with periodontal probing for the assessment of periodontal attachment of the canine teeth in dogs. STUDY POPULATION: 466 canine teeth from 117 dogs. PROCEDURE: Periodontal probing measurements were recorded, and clinical attachment levels (CAL) were calculated at the mesial, buccal, distal, and lingual (or palatal) surfaces on each canine tooth. Occlusal and lateral radiographs of the canine teeth were obtained. Alveolar margin height (AMH) was measured at the same 4 surfaces. Values for AMH and CAL were compared on the basis of tooth surface, dental arch, and radiographic view. RESULTS: The AMH at the mesial and distal surfaces of the mandibular canine teeth was measurable on the lateral view and was significantly correlated with CAL. Similar results were found for the mesial and distal surfaces of the maxillary canine teeth. Buccal and lingual AMH were measured on the mandibular occlusal radiographic view, and values were significantly correlated with CAL, but only the buccal AMH could be assessed on the occlusal radiographic view of the maxilla with values that correlated significantly with CAL. CONCLUSIONS AND CLINICAL RELEVANCE: The lateral radiographic view is suitable for evaluating periodontal attachment at the mesial and distal surfaces of the canine teeth in dogs. The occlusal radiographic view is suitable for assessing buccal surfaces as well as the lingual surface of mandibular canine teeth but not the palatal surface of maxillary canine teeth in dogs.     

<Emphasis Type="Italic">Sunflower Chlorotic Mottle Virus</Emphasis> in Compatible Interactions with Sunflower: ROS Generation and Antioxidant Response

Sunflower chlorotic mottle virus (SuCMoV) is a recently described potyvirus that causes systemic infections in sunflower plants leading to chlorotic mottling and important growth reductions and yield losses. Oxidative damage is expressed after symptom development in this host-pathogen combination. The involvement of antioxidant enzyme activities in disease susceptibility was studied in two sunflower lines differing in the intensity and rate of development of SuCMoV infections: L2 is more susceptible than L33. A transient superoxide production peak was detected in leaves of both lines before symptom development. H₂O₂ accumulation increased before symptom expression in infected plants of L33 but in L2 such increase was registered only after symptoms became evident. In healthy plants of both lines, superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) showed similar activity levels. In inoculated plants of line L2, but not in L33, SOD and CAT activities increased significantly before the appearance of symptoms, and APX increases were detected later. A 1 mM SA treatment effectively decreased SuCMoV accumulation in plants of L2 but it did not affect it in L33. This treatment increased H₂O₂ accumulation and prevented the increase in antioxidant enzyme activities in infected plants of L2. It is suggested that increases in antioxidant enzyme activities interrupted the signals generated by the increase in ROS, which may have otherwise triggered defence reactions in the host and thus, resulted in a compatible interaction.     

Paraneoplastic hypereosinophilia in a dog with intestinal T-cell lymphoma

A 9-year-old, intact male Doberman Pinscher was examined because of anorexia and weakness. Results of a CBC showed severe, microcytic, hypochromic anemia with mild eosinophilia (2944 cells/microL, reference interval 100-1250/microL) and thrombocytosis. Hypoferremia, hypoferritinemia, and a positive fecal occult blood test supported a diagnosis of iron deficiency anemia secondary to chronic intestinal hemorrhage. Abdominal ultrasound evaluation showed a thickened small intestinal loop, of which representative specimens were obtained during exploratory laparotomy. Histologically, the intestinal wall was infiltrated by a neoplastic population of large, round, lymphoid cells with vesicular chromatin, 1 or more prominent nucleoli, and a high number of mitotic figures. The cells were closely admixed with mature eosinophils, but were negative for metachromatic granules with toluidine blue. Immunohistochemically, tumor cells were positive for CD3, and negative for CD21, Pan B, and CD79a. A diagnosis of intestinal T-cell lymphoma was made. Chemotherapy was begun, with 30 mg/m;2 of doxorubicin administered intravenously every 3 weeks. Eosinophil concentration was 880/microL 2 weeks after surgery (on day 15 after presentation) but increased markedly to 62,914/microL on day 30, 62,400/microL on day 37, and 39,444/microL on day 58 after presentation. An association between hypereosinophilia and T-cell lymphoma is well established in human patients, in whom production of IL-5 by neoplastic T cells has been demonstrated. Hypereosinophilia has been reported only rarely with intestinal lymphoma in cats and horses, and with T-cell lymphoma in dogs.     

Serum biochemistry in Andean flamingos (Phoenicoparrus andinus): natural versus artificial diet.

A study of 10 clinical pathology values in four groups of Andean flamingo chicks (Phoenicoparrus andinus) was conducted to evaluate an artificial feeding program in Chile. Three groups were fed controlled diets (groups 2000, 2001, and 2002) with quantitative differences in their nutritional content. A fourth group of free-living Andean flamingo chicks was used as normal controls. Nutritional management techniques used in 2002 resulted in hematologic values with similar levels of total protein, globulins, albumin, cholesterol, urea, phosphorus, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, and calcium to those obtained in free-living chicks. In addition, final weight, physical condition, and plumage in flamingo chicks of group 2002 were considered satisfactory to face local climatic conditions and nomadic activity. These results may be useful as reference values and help to improve conservation management and veterinary care of this species.     

Suppression of seed-borne Alternaria arborescens and growth enhancement of wheat with biorational fungicides

Alternaria spp. are among the major fungal contaminants of wheat grain under postharvest and storage conditions, where A. arborescens was recently detected as a new member of this complex in Argentina causing black point. The aim of this study was to assess the potential of some biorational agents to control A. arborescens and their plant growth promoting of wheat. Seed treatments with spore suspensions of Trichoderma harzianum and Eppicoccum nigrum, extracts from Lippia alba and garlic, sodium bicarbonate, salicylic acid (SA), potassium chloride and dibasic sodium phosphate (SP) were applied to grains of wheat cultivar BIOINTA 1004 before their inoculation with the pathogen. After 7 days, seed germination and infection, necrotic symptoms on emerged seedlings and fresh weight were evaluated. Remarkable results were obtained with L. alba, SA and SP treatments that reduced symptoms markedly compared with the control. Interestingly, necrosis of radicles was significantly reduced by the application of all treatments tested. Moreover, fresh weight of seedlings was significantly increased with the application of the two antagonists, diluted garlic juice and the three tested salts in comparison with controls. Therefore, a positive role as growth promoters can be elucidated. It is concluded that compounds here tested have potential as ecofriendly alternatives to control seed-borne Alternaria fungi of wheat.