Larval Therapy and Larval Excretions/Secretions: A Potential Treatment for Biofilm in Chronic Wounds? A Systematic Review.

Chronic wounds present a global healthcare challenge and are increasing in prevalence, with bacterial biofilms being the primary roadblock to healing in most cases. A systematic review of the to-date knowledge on larval therapy's interaction with chronic-wound biofilm is presented here. The findings detail how larval therapy-the controlled application of necrophagous blowfly larvae-acts on biofilms produced by chronic-wound-relevant bacteria through their principle pharmacological mode of action: the secretion and excretion of biologically active substances into the wound bed. A total of 12 inclusion-criteria-meeting publications were identified following the application of a PRISMA-guided methodology for a systematic review. The findings of these publications were qualitatively analyzed to provide a summary of the prevailing understanding of larval therapy's effects on bacterial biofilm. A further review assessed the quality of the existing evidence to identify knowledge gaps and suggest ways these may be bridged. In summary, larval therapy has a seemingly unarguable ability to inhibit and degrade bacterial biofilms associated with impaired wound healing. However, further research is needed to clarify and standardize the methodological approach in this area of investigation. Such research may lead to the clinical application of larval therapy or derivative treatments for the management of chronic-wound biofilms and improve patient healing outcomes at a time when alternative therapies are desperately needed.

Pressure Ulcer Prevalence by Level of Paralysis in Patients With Spinal Cord Injury in Long-term Care.

OBJECTIVE: Scientific literature suggests pressure ulcer (PU) risk increases as immobility increases, indicating that more extensive paralysis confers a greater risk of PU. Yet the specific level of paralysis (ie, hemiplegia vs paraplegia vs quadriplegia), apart from neurodegenerative diagnoses, has never been examined in the long-term care (LTC) population. This study examined the prevalence of PU among LTC residents with different paralysis levels. METHODS: The authors conducted a secondary data analysis of the 2012 US Minimum Data Set of LTC facilities (n = 51,664 residents). Measures included PU stage, level of paralysis, functional impairments, comorbidities, and sociodemographic factors. After removing residents with neurodegenerative disease, comatose patients, and those with hip fractures from the analysis, logistic regressions were used to examine the association of risk factors and sociodemographic characteristics with the presence of PU. MAIN RESULTS: The sample included 7,540 patients with quadriplegia, 11,614 patients with paraplegia, and 32,510 patients with hemiplegia in LTC facilities. The PU prevalence in the sample (stages 2, 3, and 4; suspected deep-tissue injury; and unstageable PUs) was 33.9% for patients with quadriplegia, 47.4% for patients with paraplegia, and 9.6% for patients with hemiplegia. CONCLUSIONS: Within paralysis groups (quadriplegic, paraplegic, hemiplegic), risk factors for PU differed in type and magnitude. The PU rates associated with quadriplegia and paraplegia are much higher than LTC residents without paralysis, and PU prevalence for hemiplegia is similar to the rate in LTC residents without paralysis. When the risk factor of paraplegia versus quadriplegia was isolated, PU prevalence for patients with paraplegia was significantly higher.

Longitudinal Examination of Bone Loss in Male Rats After Moderate-Severe Contusion Spinal Cord Injury.

To elucidate mechanisms of bone loss after spinal cord injury (SCI), we evaluated the time-course of cancellous and cortical bone microarchitectural deterioration via microcomputed tomography, measured histomorphometric and circulating bone turnover indices, and characterized the development of whole bone mechanical deficits in a clinically relevant experimental SCI model. 16-weeks-old male Sprague-Dawley rats received T9 laminectomy (SHAM, n = 50) or moderate-severe contusion SCI (n = 52). Outcomes were assessed at 2-weeks, 1-month, 2-months, and 3-months post-surgery. SCI produced immediate sublesional paralysis and persistent hindlimb locomotor impairment. Higher circulating tartrate-resistant acid phosphatase 5b (bone resorption marker) and lower osteoblast bone surface and histomorphometric cancellous bone formation indices were present in SCI animals at 2-weeks post-surgery, suggesting uncoupled cancellous bone turnover. Distal femoral and proximal tibial cancellous bone volume, trabecular thickness, and trabecular number were markedly lower after SCI, with the residual cancellous network exhibiting less trabecular connectivity. Periosteal bone formation indices were lower at 2-weeks and 1-month post-SCI, preceding femoral cortical bone loss and the development of bone mechanical deficits at the distal femur and femoral diaphysis. SCI animals also exhibited lower serum testosterone than SHAM, until 2-months post-surgery, and lower serum leptin throughout. Our moderate-severe contusion SCI model displayed rapid cancellous bone deterioration and more gradual cortical bone loss and development of whole bone mechanical deficits, which likely resulted from a temporal uncoupling of bone turnover, similar to the sequalae observed in the motor-complete SCI population. Low testosterone and/or leptin may contribute to the molecular mechanisms underlying bone deterioration after SCI.

Nonconsumptive Effects of Predatory Chrysomya rufifacies (Diptera: Calliphoridae) Larval Cues on Larval Cochliomyia macellaria (Diptera: Calliphoridae) Growth and Development.

Forensic entomologists often rely on development data associated with a given species to estimate when it colonized human or other vertebrate remains. In most instances, these development studies are based on single species reared in isolation in the laboratory. This study examined the impact of excretions and secretions (ES) associated with third-instar Chrysomya rufifacies (Macquart), a predator, on the development of its prey, Cochliomyia macellaria (F.). Not surprisingly, Ch. rufifacies ES did not impact the development of first- or second-instar C. macellaria, which are typically not preyed on by Ch. rufifacies. However, development of third-instar C. macellaria, which do experience predation, was impacted. First, larvae were longer than those in the control (deionized water, dH2O). Filtering the ES and removing the associated bacteria and byproducts >0.2 µm dampened the previous impact observed by the unfiltered ES on third-instar C. macellaria. Second, third-instar C. macellaria treated with unfiltered ES completed pupariation 8 h quicker than the controls. Filtering the ES lessened this effect by 50%. And finally, third-instar C. macellaria treated with filtered or unfiltered Ch. rufifacies ES reached adulthood ∼5 h faster than controls treated with dH2O. In summary, these data have large ramifications for forensic entomology, as multiple species being present on decomposing remains is not uncommon. Understanding the impact of associated ES produced by interspecific cohorts on associated development could lead to more precise estimates of the minimum postmortem interval for forensic investigation of decomposing remains.

Testosterone Plus Finasteride Prevents Bone Loss without Prostate Growth in a Rodent Spinal Cord Injury Model.

We have reported that testosterone-enanthate (TE) prevents the musculoskeletal decline occurring acutely after spinal cord injury (SCI), but results in a near doubling of prostate mass. Our purpose was to test the hypothesis that administration of TE plus finasteride (FIN; type II 5α-reductase inhibitor) would prevent the chronic musculoskeletal deficits in our rodent severe contusion SCI model, without inducing prostate enlargement. Forty-three 16-week-old male Sprague-Dawley rats received: 1) SHAM surgery (T9 laminectomy); 2) severe (250 kdyne) contusion SCI; 3) SCI+TE (7.0 mg/week, intramuscular); or 4) SCI+TE+FIN (5 mg/kg/day, subcutaneous). At 8 weeks post-surgery, SCI animals exhibited reduced serum testosterone and levator ani/bulbocavernosus (LABC) muscle mass, effects that were prevented by TE. Cancellous and cortical (periosteal) bone turnover (assessed by histomorphometry) were elevated post-SCI, resulting in reduced distal femur cancellous and cortical bone mass (assessed by microcomputed tomography). TE treatment normalized cancellous and cortical bone turnover and maintained cancellous bone mass at the level of SHAM animals, but produced prostate enlargement. FIN coadministration did not inhibit the TE-induced musculoskeletal effects, but prevented prostate growth. Neither drug regimen prevented SCI-induced cortical bone loss, although no differences in whole bone strength were present among groups. Our findings indicate that TE+FIN prevented the chronic cancellous bone deficits and LABC muscle loss in SCI animals without inducing prostate enlargement, which provides a rationale for the inclusion of TE+FIN in multimodal therapeutic interventions intended to alleviate the musculoskeletal decline post-SCI.

Airworthiness testing of medical maggots.

An investigation was conducted to test and certify medicinal maggots to facilitate rapid healing of traumatic and chronic wound infections in Wounded warriors being transported onboard military aircraft. Our specific aims included (1) to test the ability of medical grade larvae to withstand the rigors of U.S. Army aeromedical certification, including tolerance to change in pressure, temperature, and humidity inside ground-based chambers; (2) to evaluate the efficacy of the medical grade larvae during a high-vibration rotary-wing medical transport flight; and (3) to gain U.S. Army aeromedical certification and U.S. Air Force safe-to-fly approval and begin the steps needed to deploy/implement the use of medicinal maggots in patient care regimes for medical airlift standard operating procedures. This report outlines the ground-based and initial air-based tests performed during the study. Maggot mortality was very low during all tests, with a mortality rate of less than 1%. Maggot growth rates in wound arenas were mixed but generally depended on temperature. Overall, the results of these tests suggest that medicinal maggots can withstand the rigors of aeromedical evacuation flights in simulated flight environments and rotary- or fixed-wing aircraft.

A metagenomic assessment of the bacteria associated with Lucilia sericata and Lucilia cuprina (Diptera: Calliphoridae).

Lucilia Robineau-Desvoidy (Diptera: Calliphoridae) is a blow fly genus of forensic, medical, veterinary, and agricultural importance. This genus is also famous because of its beneficial uses in maggot debridement therapy (MDT). Although the genus is of considerable economic importance, our knowledge about microbes associated with these flies and how these bacteria are horizontally and trans-generationally transmitted is limited. In this study, we characterized bacteria associated with different life stages of Lucilia sericata (Meigen) and Lucilia cuprina (Wiedemann) and in the salivary gland of L. sericata by using 16S rDNA 454 pyrosequencing. Bacteria associated with the salivary gland of L. sericata were also characterized using light and transmission electron microscopy (TEM). Results from this study suggest that the majority of bacteria associated with these flies belong to phyla Proteobacteria, Firmicutes, and Bacteroidetes, and most bacteria are maintained intragenerationally, with a considerable degree of turnover from generation to generation. In both species, second-generation eggs exhibited the highest bacterial phylum diversity (20 % genetic distance) than other life stages. The Lucilia sister species shared the majority of their classified genera. Of the shared bacterial genera, Providencia, Ignatzschineria, Lactobacillus, Lactococcus, Vagococcus, Morganella, and Myroides were present at relatively high abundances. Lactobacillus, Proteus, Diaphorobacter, and Morganella were the dominant bacterial genera associated with a survey of the salivary gland of L. sericata. TEM analysis showed a sparse distribution of both Gram-positive and Gram-negative bacteria in the salivary gland of L. sericata. There was more evidence for horizontal transmission of bacteria than there was for trans-generational inheritance. Several pathogenic genera were either amplified or reduced by the larval feeding on decomposing liver as a resource. Overall, this study provides information on bacterial communities associated with different life stages of Lucilia and their horizontal and trans-generational transmission, which may help in the development of better vector-borne disease management and MDT methods.

Effects of temperature and tissue type on Chrysomya rufifacies (Diptera: Calliphoridae) (Macquart) development.

The hairy maggot blow fly, Chrysomya rufifacies (Diptera: Calliphoridae), is a forensically important fly often encountered on human and other vertebrate remains in temperate and tropic regions throughout the world including Australia, Asia, Central America and North America. C. rufifacies was reared under controlled laboratory conditions on three muscle types (i.e., porcine, equine and canine) at three temperatures (i.e., 20.8, 24.8 and 28.3°C). Rate of larval weight gain across time was statistically significant between muscle types (P≤0.0001) and approaching significance across time between temperatures (P=0.0511). This research represents the first development study for C. rufifacies from central Texas, USA and the first study to examine the impact of tissue type on its development. Furthermore, these data, when compared to those available in the literature, indicate developmental differences that could be due to genetic differences in populations or possibly methods employed during the studies. Caution should be emphasized when applying development data for this species from one region to forensic investigations in other ecoregions as such differences in development based on tissue fed upon by larvae, population genetics, and methodologies used in the studies could represent error in estimating the time of colonization.

Occurrence of oriental flies associated with indoor and outdoor human remains in the tropical climate of north Malaysia.

Flies attracted to human remains during death investigations were surveyed in north Peninsular Malaysia. Six families, eight genera, and 16 species were identified from human remains, with the greatest fly diversity occurring on remains recovered indoors. The total relative frequency of species was led by Chrysomya megacephala (Fabricius, 1794) (46%), followed by Chrysomya rufifacies (Macquart, 1842) (22%), Sarcophaga (Liopygia) ruficornis (Fabricius, 1974) (5%), Sarcophaga spp. (4%), Synthesiomyia nudiseta Wulp, 1883 (6%), Megaselia spp. (3%), Megaselia scalaris (Loew, 1866), (2%), Megaselia spiracularis Schmitz, 1938 (2%), and Chrysomya villeneuvi Patton, 1922 (2%). Hemipyrellia tagaliana (Bigot, 1877), Desmometopa sp., Megaselia curtineura (Brues, 1909), Hemipyrellia ligurriens Wiedemann 1830, Ophyra sp., Sarcophaga princeps Wiedemann 1830, Piophila casei (Linnaeus, 1758), and unidentified pupae each represented 1%, respectively.