Identification of unique DNA sequences present in highly virulent 2009 Alabama isolates of Aeromonas hydrophila

In 2009, a disease outbreak caused by Aeromonas hydrophila occurred in 48 catfish farms in West Alabama, causing an estimated loss of more than 3 million pounds of food size channel catfish. Virulence studies have revealed that the 2009 isolates of A. hydrophila are at least 200-fold more virulent than a 1998 Alabama isolate AL98-C1B. However, up to now, no molecular markers have been identified to differentiate the highly virulent 2009 isolates from other isolates of A. hydrophila. To understand the genetic differences between the highly virulent 2009 isolates and the less virulent AL98-C1B at molecular level, PCR-select bacterial genome subtractive hybridization was used in this study. A total of 96 clones were selected from the subtractive genomic DNA library. Sequencing results revealed that the 96 clones represented 64 unique A. hydrophila sequences. Of the 64 sequences, three (hypothetical protein XAUC_13870, structural toxin protein RtxA, and putative methyltransferase) were confirmed to be present in the three virulent 2009 Alabama isolates but absent in the less virulent AL98-C1B. Using genomic DNAs from nine field isolates of A. hydrophila with different virulence as templates, two sequences (hypothetical protein XAUC_13870 and putative methyltransferase) were found to be only present in highly virulent A. hydrophila isolates, but absent in avirulent isolates.     

Bovine viral diarrhoea virus (BVDV) subgenotypes in diagnostic laboratory accessions: distribution of BVDV1a, 1b, and 2a subgenotypes

The prevalence of bovine viral diarrhoea virus (BVDV) biotypes and subgenotypes was determined from 131 BVDV positive samples from a diagnostic laboratory. The majority of the isolates were from Oklahoma; however, other states including Kansas, Texas, and Arkansas were represented. These BVDV samples were from submissions of 76 live animals and 55 necropsy samples. There were 131 BVDV samples represented by 117 noncytopathic (NCP), 11 cytopathic (CP) and 3 cases with mixed NCP and CP biotypes. The NCP isolates were more common (P < 0.05) than the CP and NCP/CP combination. The BVDV samples were segregated into three subgenotypes by differential PCR and sequencing of a viral genomic region, 5'-untranslated region (5'-UTR). There were more BVDV1b subgenotypes 60/131 (45.8%) than BVDV1a, 37/131 (28.2%) or BVDV2a, 34/131 (26.0%) (P < 0.05). The organ system involvement included the major categories such as respiratory, digestive, mixed/multiple organs, abortions, and persistent infections (PI). All three BVDV subgenotypes were found in persistently infected (PI) cattle and respiratory diseases, both major requests for BVDV diagnosis. Only one of the 131 viruses was genetically similar to the strains present in U.S. vaccines.     

Lindane Biodegradation by Defined Consortia of Indigenous Streptomyces Strains

The current study aimed to compare lindane degradation by pure and mixed cultures of Streptomyces sp. Cell-free extracts were assayed for potentiating dechlorinase activity and, based on these results, consortia of two to six microorganisms were assayed for their growth on and degradation of lindane. Furthermore, the role of bacterial consortia of lindane-degrading strains was examined in lindane decontamination soil assays. Four actinobacteria, previously isolated from a pesticide-contaminated area, were selected because of their tolerance to lindane and their ability to use the pesticide as sole carbon source. These strains as well as Streptomyces sp. M7 and Streptomyces coelicolor A3 were used to study specific dechlorinase activity (SDA) and lindane removal in mixed cultures. Pure cultures presented SDA in the presence of 1.66 mg L^-1 lindane as carbon source. SDA was improved by certain mixed cultures until 12 times compared with pure cultures. Mixed cultures with two, three, and four strains showed maximum lindane removal of 46% to 68%, whereas combinations of five and six strains did not efficiently remove the pesticide from the culture medium. The Streptomyces sp. A2, A5, M7, and A11 consortium presented the lowest ratio between residual lindane concentration and SDA and could be a promising tool for lindane biodegradation.     

Functional genomics of microspore embryogenesis

Isolated plant microspores, when stressed and cultured in vitro, can be diverted from their normal gametophytic pathway towards sporophytic development, with the formation of haploid embryos and ultimately doubled-haploid plants. This process is called androgenesis or microspore embryogenesis, and is widely used in plant breeding programmes to generate homozygous lines for breeding purposes. Protocols for the induction of microspore embryogenesis and the subsequent regeneration of doubled haploid (DH) plants have been successfully developed for more than 200 species. These practical advances stand in stark contrast to our knowledge of the underlying molecular genetic mechanism controlling this process. The majority of information regarding the genetic and molecular control of the developmental switch from gametophytic to sporophytic development has been garnered from four intensely studied (crop) plants comprising two dicotyledonous species, rapeseed (Brassica napus) and tobacco (Nicotiana tabacum), and two monocotyledonous species, wheat (Triticum aestivum) and barley (Hordeum vulgare). In these species the efficiency of microspore embryogenesis is very high and reproducible, making them suitable models for molecular studies. In the past, molecular studies on microspore embryogenesis have focussed mainly on the identification of genes that are differentially expressed during this developmental transition and/or early in embryo development, and have identified a number of genes whose expression marks or predicts the developmental fate of stressed microspores. More recently, functional genomics approaches have been used to obtain a broad overview of the molecular processes that take place during the establishment of microspore embryogenesis. In this review we summarise accumulated molecular data obtained in rapeseed, tobacco, wheat and barley on embryogenic induction of microspores and define common aspects involved in the androgenic switch.     

Questionnaire on the process of recovering horses from general anesthesia and associated personnel injury in equine practice

ObjectiveTo collect data about the current practice of recovering horses from general anesthesia and recovery personnel safety.Study designOnline survey.MethodsAn online questionnaire, including questions on general demographic data, recovery drugs, modality and characteristics of equine recovery and morbidity and mortality, was designed and distributed via e-mail to equine practitioners worldwide.ResultsPractitioners from 22 countries completed 373 questionnaires; 53% of the participants were board-certified equine surgeons, and the remainder were board-certified anesthesiologists (18%), large animal residents (8%), general practitioners (7%), large animal interns (6%), anesthesia residents (4.5%) and veterinary technicians (1.6%). Respondents were employed by academia (58%) or private practice (42%). Of the respondents employed at a university, 93% had a board-certified anesthesiologist on staff compared with 7% of respondents employed at a private practice. Most of the respondents assist horses during recovery, with 23% assisting every recovery and 44% assisting recovery in the majority of cases. Reasons for choosing to assist horses during recovery were: orthopedic procedures (57%), neurological deficits (49%), bad health (47%), history of poor recovery (44%), foals (42%), draft breeds (30%), magnetic resonance imaging (17%) and computed tomography (16%). Unacceptable recoveries were reported by 77% of participants. Commonly reported complications during recovery with any method were: orthopedic injury (66%), myopathy (54%), skin abrasion (53%) and airway obstruction (37%). The incidences of unacceptable quality of recovery (p = 0.09) or personnel injury (p = 0.56) were not different between assisted and nonassisted recoveries; however, more equine fatalities were reported for assisted recoveries (p < 0.006). Practitioners in academia reported more unacceptable recoveries (p < 0.0007) and personnel injuries (p < 0.002) compared with those in private practice.ConclusionsThe method of recovery differs among hospitals. Recovery personnel injuries associated with assisting horses during recovery are an important and previously unreported finding.     

Production of Surface-Active Compounds by a Hydrocarbon-Degrading Actinobacterium: Presumptive Relationship with Lipase Activity

The replacement of synthetic surface-active compounds (SACs) by their microbial counterparts is carving out a niche for themselves in the field of bioremediation. However, the high cost of microbial products has limited their application at a realistic scale. In the current study, several hydrocarbon-degrading microorganisms were assayed as potential SAC producers in low-cost liquid media. Only the strain CC10, placed within the class Actinobacteria, was able to produce emulsifying molecules by using a combination of sugarcane vinasse or crude glycerol (as cheap carbon substrates) with urea or peptone (as nitrogen sources). The emulsifying activity of the supernatants and the stability of emulsions formed with motor oil depended on the carbon and nitrogen sources. However, the biodegradability of these metabolites was only associated with the carbon substrate, and it was always higher than the two tested synthetic surfactants, sodium dodecyl sulfate and Triton X-100. Also, a positive linear association between emulsifying and lipase activities of the CC10 supernatants was detected (r?=?0.781; p?=?0.219), with the maximum activities detected in the glycerol-peptone supernatant. Interestingly, this supernatant was able to emulsify different oily substrates, a property that could be used to increase the efficiency of the treatment of effluents with high fat content.     

Leptin regulates striatal regions and human eating behavior

Studies of the fat-derived hormone leptin have provided key insights into the molecular and neural components of feeding behavior and body weight regulation. An important challenge lies in understanding how the rewarding properties of food interact with, and can override, physiological satiety signals and promote overeating. We used functional magnetic resonance imaging to measure brain responses in two human patients with congenital leptin deficiency who were shown images of food before and after 7 days of leptin replacement therapy. Leptin was found to modulate neural activation in key striatal regions, suggesting that the hormone acts on neural circuits governing food intake to diminish the perception of food reward while enhancing the response to satiety signals generated during food consumption.     

Compartmentalized control of skin immunity by resident commensals

Intestinal commensal bacteria induce protective and regulatory responses that maintain host-microbial mutualism. However, the contribution of tissue-resident commensals to immunity and inflammation at other barrier sites has not been addressed. We found that in mice, the skin microbiota have an autonomous role in controlling the local inflammatory milieu and tuning resident T lymphocyte function. Protective immunity to a cutaneous pathogen was found to be critically dependent on the skin microbiota but not the gut microbiota. Furthermore, skin commensals tuned the function of local T cells in a manner dependent on signaling downstream of the interleukin-1 receptor. These findings underscore the importance of the microbiota as a distinctive feature of tissue compartmentalization, and provide insight into mechanisms of immune system regulation by resident commensal niches in health and disease.