Novel Alkaloids from Marine Actinobacteria: Discovery and Characterization

The marine environment is an excellent resource for natural products with therapeutic potential. Its microbial inhabitants, often associated with other marine organisms, are specialized in the synthesis of bioactive secondary metabolites. Similar to their terrestrial counterparts, marine Actinobacteria are a prevalent source of these natural products. Here, we discuss 77 newly discovered alkaloids produced by such marine Actinobacteria between 2017 and mid-2021, as well as the strategies employed in their elucidation. While 12 different classes of alkaloids were unraveled, indoles, diketopiperazines, glutarimides, indolizidines, and pyrroles were most dominant. Discoveries were mainly based on experimental approaches where microbial extracts were analyzed in relation to novel compounds. Although such experimental procedures have proven useful in the past, the methodologies need adaptations to limit the chance of compound rediscovery. On the other hand, genome mining provides a different angle for natural product discovery. While the technology is still relatively young compared to experimental screening, significant improvement has been made in recent years. Together with synthetic biology tools, both genome mining and extract screening provide excellent opportunities for continued drug discovery from marine Actinobacteria.     

Real-time polymerase chain reaction testing for the detection of Mycobacterium genavense and Mycobacterium avium complex species in avian samples

Diagnosis of avian mycobacteriosis, caused by Mycobacterium genavense or species belonging to the Mycobacterium avium complex (MAC), is problematic. Polymerase chain reaction (PCR) offers rapid and sensitive detection of minute quantities of DNA, and conventional protocols have been used for evaluating avian specimens. The recent development of real-time PCR offers several advantages over conventional PCR. In attempts to improve diagnosing avian mycobacteriosis, a real-time TaqMan PCR assay was developed targeting the 65-kD heat shock protein gene of M. genavense and MAC spp. Nineteen reference isolates, 16 clinical isolates, and 32 avian tissue samples were used to evaluate the assay. When sufficient amplicons were produced, the species of mycobacteria was determined by standard sequencing of TaqMan PCR products and compared with results from commercial mycobacteriology laboratories and/or standard sequencing of conventional PCR products. The TaqMan PCR detected DNA from reference isolates of M. genavense, MAC spp., and Mycobacterium tuberculosis complex spp. Of the clinical isolates, the TaqMan PCR detected DNA from 10 of 12 Mycobacterium avium avium isolates and two of three Mycobacterium avium intracellulare isolates. For the tissue samples, the TaqMan PCR amplified DNA in six of nine samples that were identified by sequencing of conventional PCR products and/or by commercial mycobacteriology laboratories as being MAC spp. positive and three of four samples that were positive for M. genavense. There was some disagreement between speciation results from the TaqMan PCR and those from commercial mycobacteriology laboratories or conventional PCR or both. This disagreement was suspected to be because of relatively small numbers of base pairs in the TaqMan PCR products. The TaqMan PCR may provide a useful tool for evaluating clinical samples for DNA from mycobacteria species that most commonly infect birds; however, further refinement is needed in order to improve sensitivity and provide more accurate speciation.     

Ultrasonographic and histologic evaluation of medial and middle patellar ligaments in exercised horses following injection with ethanolamine oleate and 2% iodine in almond oil

OBJECTIVE: To describe the ultrasonographic and quantitative histologic effect of injecting 2% iodine in almond oil (IAO) and ethanolamine oleate (EO) in the medial and middle patellar ligaments of horses and to determine whether a difference in response exists between IAO and EO treatment. ANIMALS: 10 healthy horses. PROCEDURE: In 5 horses, the medial and middle patellar ligaments of 1 limb were injected with EO, whereas IAO was injected in the medial and middle patellar ligaments of another 5 horses. Ultrasonographic evaluation was performed on the experimental and control limb before injection of IAO and EO and prior to euthanasia to determine cross-sectional area and evaluate fiber pattern. The patellar ligaments were harvested 2 weeks after injection and examined histologically to evaluate the inflammatory response, fibroplasia, and chondroid metaplasia. RESULTS: Injection of the patellar ligaments with IAO resulted in a greater increase in cross-sectional area on ultrasonography than EO. Both agents caused a decrease in echogenicity of the ligament. Histologically, significantly greater infiltration of inflammatory cells and fibroplasia developed after injection with IAO, compared with EO. Both agents resulted in significantly greater fibroplasia relative to control specimens. CONCLUSIONS AND CLINICAL RELEVANCE: Injection of the medial and middle patellar ligaments with IAO induces more severe inflammation and fibroplasia than EO. Maturation of the inflammatory and fibrous response may contribute to resolution or attenuation of upward fixation of the patella by subsequent stiffening of the ligaments.     

Assessing performance of feedlot operations using epidemiology.

The progressive feedlot veterinarian must be well versed not only in individual production animal medicine, but also in population-based medicine. Feedlot health programs must be goal oriented, and evaluation of these goals is accomplished through diligent use of record systems and analytic evaluation of these record systems. Basic feedlot monitoring parameters include health and economic parameters in addition to the use of bench marking parameters between and among feed yards. When these parameters have significant changes, steps should be initiated to begin field investigations. Feedlot epidemiology uses several novel applications such as partial budgeting, risk assessment, and packing plant audits to provide scientifically sound and economically feasible solutions for the feeding industry.     

Concurrent West Nile virus and mycobacterium avium infection in a black-necked swan (Cygnus melanocoryphus).

An adult male black-necked swan (Cygnus melanocoryphus) was evaluated for a routine physical examination and West Nile virus vaccination. Nine days later, the swan was hospitalized for weakness, dehydration, tremors, and leukocytosis. Gradual clinical improvement was seen over a 2-wk course of supportive care. However, the animal became acutely weak and was found dead. Histopathology revealed severe granulomatous pneumonia and air sacculitis with dissemination to the liver, spleen, intestine, and coelom. Mycobacterial infection was confirmed by identification of acid-fast bacilli within tissue sections and by polymerase chain reaction. Concurrent West Nile virus infection was identified in sections of brain by using immunohistochemistry.     

New techniques for breeding maize (Zea mays) varieties with fall armyworm resistance and market-preferred traits for sub-Saharan Africa

Deploying maize varieties with fall armyworm (Spodoptera frugiperda [J.E. Smith]; FAW) resistance, desirable product profiles (PPs) and climate resilience is fundamental for food and economic security in sub-Saharan Africa (SSA). This study reviewed and identified challenges and opportunities for effective and accelerated breeding of demand-led maize hybrids with FAW resistance and adaptation to the diverse agro-ecologies of SSA. Lessons were drawn on improving breeding efficiency through adequate genetic variation delivered via prebreeding programmes, speed breeding and a reduced breeding stage plan. Appropriate PPs aligned with demand-led breeding approaches were highlighted as foundations for variety design and commercialization. Challenges to accelerated FAW resistance breeding in maize included inadequate funds and modern tools; poor adaptation of some exotic donor parental lines; lack of information on FAW resistance among local varieties; lack of integration of molecular markers associated with FAW resistance and agronomic traits into selection plans; and limited infrastructure for FAW rearing and germplasm screening. Integration of modern breeding tools and scientific innovations were recommended for accelerated development and release of FAW resistant and market-preferred maize varieties.     

Color stability, reducing activity, and cytochrome c oxidase activity of five bovine muscles

The objective was to characterize the beef psoas major (PM), longissimus lumborum (LL), superficial semimembranosus (SSM), deep semimembranosus (DSM), and semitendinosus (ST) muscles for differences in instrumental and visual color, metmyoglobin-reducing activity (MRA), total reducing activity (TRA), and cytochrome c oxidase activity. The LL and ST had the most color stability and MRA (p < 0.05), the DSM and PM had the least (p < 0.05), and values for the SSM were intermediate. Visual color (r = -0.66) and a and chroma (r = 0.68) were more correlated with MRA than with TRA (r < 0.14 for all measures). This research supports previous reports that color stability among muscles is variable and that MRA is more useful than TRA for explaining the role of reducing activity in muscle-color stability.     

Surgical amputation of a digit and vacuum-assisted-closure (V.A.C.) management in a case of osteomyelitis and wound care in an eastern black rhinoceros (Diceros bicornis michaeli)

A 14-yr-old female eastern black rhinoceros (Diceros bicornis michaeli) presented with progressive suppurative osteomyelitis in her left hind lateral toe. beta-Hemolytic Streptococcus sp. was isolated. The animal was treated with multiple systemic antibiotics, and topical wound cleansing. Repeated debridements and nail trimmings were performed for 5 mo prior to electing amputation. The toe was surgically amputated under general anesthesia between the first and second phalanges. Analgesia was diffused into the wound topically via a catheter and elastomeric pump. The open amputation site was covered with adherent drapes and a negative-pressure wound therapy device provided vacuum-assisted closure (V.A.C.) for 72 hr. Three months later this animal developed a deep dermal ulcer on the lateral aspect of the right hind limb, at the level of the stifle. Methicillin-resistant Staphylococcus aureus was isolated. The wound was managed by initial daily lavage, followed by 1 mo of V.A.C. therapy, with 72 hr between dressing changes. Clinically, this therapy expedited the formation of healthy granulation tissue and overall healing was accelerated. The animal tolerated the machine and bandage changes well via operant conditioning. The use of negative-pressure wound therapy appeared to shorten time to resolution of slow-healing wounds in black rhinoceros.