Survey of state animal health officials reveals lack of personnel and resources to manage equine infectious disease outbreaks.

OBJECTIVE: The purpose of the study was to assess the challenges faced by state animal health officials (SAHOs) when managing equine disease outbreaks. METHODS: A survey was conducted to assess SAHOs' collaboration with horse owners and practicing veterinarians to provide biosecurity and infection disease control. It was sent to 180 email addresses listed in the Equine Disease Communication Center website database. The responses in each category for each question were tabulated and reported as a percentage. RESULTS: There were 72 survey responses from 45 states. The number of states responding suggested that the results represent SAHOs' current opinions. Fifty percent of responders identified the lack of resources and training for equine health management and equine reportable disease prevention and control. There is a lack of owner compliance in managing disease outbreaks, and boarding stables, horse shows, and racing facilities require the most resources for outbreak management. The survey indicated on-site communication as the most effective for managing an outbreak, and the Equine Disease Communication Center is to be a valued resource for alerts and disease information. CONCLUSIONS: At the state level, there is a lack of needed personnel and resources to effectively manage an equine infectious disease outbreak. Furthermore, failure of veterinary and owner compliance in reporting and managing equine infectious diseases allows disease spread. CLINICAL RELEVANCE: Lack of sufficient resources for management of reportable diseases will potentially result in disease spread with a negative impact on horse health and the equine industry.

Trainee educational curriculum to standardize central venous catheter repair.

INTRODUCTION: Children may require durable central venous catheters (CVCs) for various reasons. CVC-related integrity complications are common and can often be repaired at the bedside to increase lifetime. Variability in repair techniques can lead to complications, including the need for repeat repair and Central Line Associated Blood Stream Infection (CLABSI). METHODS: The impact of an educational curriculum to standardize tunneled CVC repairs for trainees on a pediatric surgery service was studied, focusing on comfort level with tunneled CVC repair and to determine the impact on complication rates. Rotating trainees studied a dedicated audiovisual educational curriculum comprised of a video, educational slides, and a practical component from November 2020 through January 2022. Experience and comfort level with tunneled CVC repairs were assessed before and after the rotation. CVCs repaired during the duration of the study were evaluated and compared to the period prior. RESULTS: Forty-nine individuals completed the pre- and post-training survey. Respondents (34.7%, n = 17) most commonly reported one year of surgical experience, and (79.6%, n = 39) had never observed or assisted in a repair previously. Following training, respondents felt more comfortable with all aspects of the CVC repair process (p < 0.001). There were no statistically significant differences in re-repair rates or CLABSI rates following the implementation of the curriculum. CONCLUSIONS: Tunneled CVC procedural repair variability can be standardized with a dedicated educational curriculum for rotating trainees, which improves knowledge and comfort with such procedures.

Pharmacokinetics of hydroxyzine and cetirizine following oral administration of hydroxyzine to exercised Thoroughbred horses.

Hydroxyzine is a first-generation antihistamine and cetirizine, a second-generation antihistamine and active metabolite of hydroxyzine. Hydroxyzine is commonly used in performance horses and as such its use in closely regulated; however, there are no published studies suitable for establishing appropriate regulatory recommendations. In the current study, 12 exercised Thoroughbred research horses received a single oral administration of 500 mg of hydroxyzine. Blood and urine samples were collected prior to and up to 96 hr postdrug administration and concentrations of hydroxyzine and cetirizine determined using liquid chromatography-tandem mass spectrometry. A joint parent/metabolite population 2-compartment pharmacokinetic model with first-order absorption and elimination was utilized to describe the pharmacokinetics of both compounds. Serum hydroxyzine and cetirizine concentrations were above the limit of quantitation (0.1 ng/ml) of the assay at 96 hr (the last time point sampled). The terminal half-life was 7.41 and 7.13 hr for hydroxyzine and cetirizine, respectively. Findings from this study suggest that a prolonged withdrawal time should be observed if this compound is used in performance administered to performance horses and is classified as prohibited substance by the applicable regulatory body.

Hormones and their Structural and Functional Effects on the Brain: How Can We Change our Practice Moving Forward?

Is hormone treatment an invasive procedure? In this paper, we discuss aspects related to the choice of treating disorders of sex development (DSD) using hormones. Specifically, we focus on some of the challenging issues related to this treatment and the need to establish a standard of care for the use of hormone therapy in this patient population. The objectives of this paper are to: 1) Enhance understanding of the uncertainties in the decision-making process regarding hormonal interventions to treat patients with DSD. 2) Recognize that the effects of hormonal interventions might require a consent process similar to that applied for surgical procedures. 3) Emphasize the need to establish treatment algorithms that could form the basis of a standard of care for this patient population.

Clinical and Radiographic Outcomes After Submuscular Plating (SMP) of Pediatric Femoral Shaft Fractures.

BACKGROUND: To evaluate lower extremity strength and alignment among children who underwent submuscular plating (SMP). METHODS: Subjects who underwent SMP for a length unstable femoral fracture returned for isokinetic strength testing ≥2 years after surgery. Extensor and flexor strength deficits (percent difference) between the operative and nonoperative limbs were evaluated. Radiographic measurements of mechanical alignment [anatomic lateral distal femoral angle (aLDFA)] and qualitative measurements (The Pediatric Outcomes Data Collection Instrument, PODCI) were obtained from all subjects. The upper 97.5% confidence interval approach to establishing clinical equivalency was utilized to compare differences in strength and alignment between the operative and nonoperative limbs. An extensor strength deficit of >19% and an aLDFA discrepancy of >5 degrees were considered to be clinically significant. RESULTS: The average age at surgery of the 10 subjects included in the study was 8.7 years. The hardware was placed an average of 27.9 mm from the distal femoral physis and was removed 6.4 months postsurgery. Among all subjects, the median PODCI scores were ≥97 according to all subscales. There was no significant difference in extension torque between the operative versus nonoperative limbs at 60 degrees/s (P=0.5400), 120 degrees/s (P=0.4214), or 180 degrees/s (P=0.8166). More importantly, extension strength deficits between the operative and nonoperative limbs were not clinically significant at 60 degrees/s [upper 97.5% confidence interval (CI), 10.9%], 120 degrees/s (upper 97.5% CI, 11.0%), or 180 degrees/s (upper 97.5% CI, 10.7%). The difference in aLDFA between the operative and nonoperative limb was less than the predefined clinically significant threshold of 5 degrees for all subjects. CONCLUSIONS: SMP achieves satisfactory clinical and functional results. In this series, extensor strength deficits and/or lower extremity malalignment were not clinically meaningful. High patient satisfaction can be expected after implant removal. LEVEL OF EVIDENCE: Level IV-case series.

Outcomes of Isolated Varus Derotational Osteotomy in Children With Cerebral Palsy Hip Dysplasia and Predictors of Resubluxation.

BACKGROUND: The appropriate intervention for hip subluxation or dislocation in children affected by cerebral palsy (CP) remains controversial. The purpose of this retrospective study was to report radiographic and clinical outcomes following isolated femoral varus derotational osteotomy (VDRO) in children with CP hip dysplasia. Risk factors for resubluxation and avascular necrosis (AVN) were also examined. METHODS: A cohort of 100 patients (199 hips) with CP treated with isolated VDRO between 2003 and 2009 was reviewed. All but 1 patient received bilateral surgery. Patients were followed for an average of 5.4 years (range, 1.03 to 10.20 y). Anteroposterior pelvic radiographs were used to assess migration percentage (MP), Shenton's line, and presence of AVN. Resubluxation was defined as a postoperative break in Shenton's line. Radiographic outcomes and risk analysis was performed in the 91 subjects (179 hips) with radiographic follow-up >1 year. RESULTS: Significant improvement was observed in MP, and all hips had a reconstituted Shenton's line following surgery. Over the course of follow-up, 16% of hips were noted to have a repeat break in Shenton's line. Univariate risk analysis showed preoperative MP, Gross Motor Function Classification System (GMFCS) level, and age at surgery were risk factors for a recurrent line break. Preoperative MP and GMFCS level were found to be predictors of resubluxation in multivariate analysis. AVN was detected in 10 hips (5.7%). GMFCS level V patients were more at risk for resubluxation, but less at risk for AVN when compared with ambulatory (GMFCS I/II/II) patients and GMFCS level IV patients. CONCLUSIONS: Performing a VDRO without additional procedures provided a stable and concentrically reduced hip joint in this population of children with CP. Attention should be paid to initial ambulatory status during the postoperative period. Concomitant procedures such as pelvic osteotomy should be considered for patients of GMFCS level IV and V, as these patients were more at risk for recurrent subluxation. LEVEL OF EVIDENCE: Level III-retrospective comparative study.

Comparison of Allograft and Bovine Xenograft in Calcaneal Lengthening Osteotomy for Flatfoot Deformity in Cerebral Palsy.

BACKGROUND: The Evan's calcaneal lengthening osteotomy is a treatment method for spastic flatfoot deformity in patients with cerebral palsy that fail nonoperative measures. Autograft and allograft have been reported as potential graft choices. Bovine xenograft has been introduced as an alternative, but limited human data exists supporting its efficacy. This study compares the long-term results of allograft versus xenograft in isolated Evan's procedure performed for correction of flexible spastic flatfoot deformity. METHODS: This retrospective study accessed charts of 4- to 18-year-olds diagnosed with cerebral palsy who received an Evan's procedure. Preoperative and postoperative radiographic measurements (lateral calcaneal pitch, lateral talocalcaneal, lateral talo-first metatarsal, anteroposterior talonavicular coverage, anteroposterior talo-first metatarsal), graft incorporation, recurrence, secondary procedures, and complications were recorded and analyzed between graft types. RESULTS: Sixty-three feet (34 allograft and 29 xenograft) in 36 patients (mean age 9.3 y) were included. Gross Motor Function Classification System between groups was significant (P=0.001). Mean time for preoperative x-rays was 5.3 months before day of surgery (DOS) for allograft and 3.6 months for xenograft. Mean time of first and last postoperative x-ray for allograft was 3.6 and 39.5 months, respectively; for xenograft, 1.8 and 35.1 months, respectively. There was a significant difference in timing of preoperative x-ray to DOS and DOS to first postoperative x-ray (P=0.012, 0.006, respectively). Radiographically, xenograft retained postoperative improvement better than allograft, yet allograft had a higher grade 4 incorporation rate (P=0.036). The allograft group experienced significantly more cast pressure ulcers (P=0.006), but no other differences in complications between groups, and no infections were reported in either group. CONCLUSIONS: Allograft incorporated better than xenograft, likely with a greater potential to reach grade 5 incorporation, yet both groups retained postoperative improvement. Results indicate both grafts are appropriate; yet incorporation rate could affect correction maintenance, and should be considered during graft selection for Evan's procedure. LEVEL OF EVIDENCE: This study presents clinical results using a novel bone graft material. Level III-retrospective comparative study.

Imaging activity in astrocytes and neurons with genetically encoded calcium indicators following in utero electroporation.

Complex interactions between networks of astrocytes and neurons are beginning to be appreciated, but remain poorly understood. Transgenic mice expressing fluorescent protein reporters of cellular activity, such as the GCaMP family of genetically encoded calcium indicators (GECIs), have been used to explore network behavior. However, in some cases, it may be desirable to use long-established rat models that closely mimic particular aspects of human conditions such as Parkinson's disease and the development of epilepsy following status epilepticus. Methods for expressing reporter proteins in the rat brain are relatively limited. Transgenic rat technologies exist but are fairly immature. Viral-mediated expression is robust but unstable, requires invasive injections, and only works well for fairly small genes (<5 kb). In utero electroporation (IUE) offers a valuable alternative. IUE is a proven method for transfecting populations of astrocytes and neurons in the rat brain without the strict limitations on transgene size. We built a toolset of IUE plasmids carrying GCaMP variants 3, 6s, or 6f driven by CAG and targeted to the cytosol or the plasma membrane. Because low baseline fluorescence of GCaMP can hinder identification of transfected cells, we included the option of co-expressing a cytosolic tdTomato protein. A binary system consisting of a plasmid carrying a piggyBac inverted terminal repeat (ITR)-flanked CAG-GCaMP-IRES-tdTomato cassette and a separate plasmid encoding for expression of piggyBac transposase was employed to stably express GCaMP and tdTomato. The plasmids were co-electroporated on embryonic days 13.5-14.5 and astrocytic and neuronal activity was subsequently imaged in acute or cultured brain slices prepared from the cortex or hippocampus. Large spontaneous transients were detected in slices obtained from rats of varying ages up to 127 days. In this report, we demonstrate the utility of this toolset for interrogating astrocytic and neuronal activity in the rat brain.

Bioinspired hybrid materials from spray-formed ceramic templates.

Thermally sprayed ceramics, when infiltrated with polymer, exhibit synergistic increases in strength and toughness. The structure of such composites-a dense, brick-mortar arrangement-is strikingly similar to that of nacre, as are the mechanisms underlying the robust mechanical behavior. This industrial-scale process thus presents an exciting tool for bio-mimetic exploration.

Burrowing in the freshwater mussel Elliptio complanata is sexually dimorphic and feminized by low levels of atrazine.

The widely used herbicide atrazine (ATR) may have endocrine-associated adverse effects, including on behavior. In this study, 120 adult freshwater mussels, Elliptio complanata, were exposed to ATR at the environmentally relevant concentrations of 1.5, 15, or 150 µg/L. Burrowing depth was evaluated hourly for 6 h and at sacrifice animals were sexed by gonad smear. Female controls burrowed overall approximately 30% less than males, the first report of sexual dimorphism in this behavior. Atrazine at 15 µg/L feminized burrowing in both sexes, in that exposed animals burrowed 20% less than their same-sex controls. Males treated with 1.5 µg /L ATR displayed approximately 20-fold higher vitellogenin (VTG) levels than same-sex controls. Higher concentrations of ATR were not associated with increasing effects. A scatterplot showed a weak binomial curve associating low burrowing with high VTG levels. Taken together, these data suggest a nonlinear dose response in behavioral and physiological feminization produced by ATR and support the need to reconsider the widespread use of this compound.

The role of gait analysis in treating gait abnormalities in cerebral palsy.

Individuals with cerebral palsy (CP) cannot take a normal activity like walking for granted. CP is the most common pediatric neurologic disorder, with an incidence of 3.6 per 1000 live births. The current trend in the treatment of individuals with CP is to perform a thorough evaluation including a complete patient history from birth to present, a comprehensive physical examination, appropriate radiographs, consultation with other medical specialists, and analysis of gait.

Decreased complications of pediatric femur fractures with a change in management.

BACKGROUND: Elastic intramedullary nails are commonly the preferred treatment option for operative stabilization of pediatric diaphyseal femur fractures. Increased complication rates have been reported in unstable fractures and older patients treated with TENs (titanium elastic nails). The reported complications have led to a change in management at our institution: limiting the use of TENs and using submuscular plating and trochanteric entry nails as alternatives. The purpose of this study is to analyze whether this change in management has improved outcomes defined by a decrease in complications between 2 time periods. METHODS: This retrospective study compared 2 cohorts of femur fractures: those treated from January 2001 to January 2003 versus those treated from January 2003 to December 2006. Patient's age, weight, fracture type (stable or unstable), operative fixation technique, and complications were compared. Outcomes were measured by major or minor complications that occurred after operative treatment. RESULTS: Period I consisted of 46 patients and Period II of 95 patients. There was a significant decrease in TEN use in unstable fractures in Period II versus Period I. Submuscular plating increased from 9% in Period I to 28% in Period II. All complications decreased from 52% in Period I to 23% in Period II. Major complications decreased from 22% to 5%, and minor complications decreased from 30% to 18%. Complications in unstable fractures decreased from 57% to 26% and in stable fractures from 48% to 22%. CONCLUSIONS: Outcomes of pediatric femur fractures are improved with limiting the use of TENs to stable fractures. Complications resulting from unstable femur fracture management have decreased with other methods of stabilization such as submuscular plating or trochanteric entry nails.

Few effects of multi-generational dietary exposure to genistein or nonylphenol on sodium solution intake in male and female Sprague-Dawley rats.

Previous work in our laboratory indicated that lifelong dietary exposure to estrogen-like endocrine disrupters increased sodium solution intake in adult male and female rats. Here, we sought to discern the critical periods necessary for this alteration as well as establish the effects of lower dietary concentrations of genistein and nonylphenol. Male and female Sprague-Dawley rats (F0) consumed phytoestrogen-free chow containing 0, 5, 100, or 500 ppm genistein (approximately equal to 0.0, 0.4, 8.0, and 40.0 mg/kg/day) or 0, 25, 200, or 750 ppm nonylphenol (approximately equal to 0.0, 2.0, 16.0, and 60.0 mg/kg/day). Rats were mated within treatment groups and offspring (F1) maintained on the same diets. Mating for the F1, F2, and F3 (genistein only) was within treatment groups. At postnatal day (PND) 21, the F3 generation began to consume unadulterated phytoestrogen-free chow such that genistein exposure occurred only in utero and preweaning. The F4 generation was never directly exposed to genistein. On PNDs 65-68, intake of regular water and a 3.0% sodium chloride solution was measured for F1-F4 generations (genistein portion) or F1-F2 (nonylphenol portion). Although body weights were decreased by the highest dietary concentrations of genistein and nonylphenol, there were only minimal effects of exposure on sodium solution intake. As expected, intake was highest in female rats. With previous data, these results indicate that the dietary concentrations necessary to increase adult sodium solution intake in rats are greater than 500 ppm genistein and 750 ppm nonylphenol and such effects do not appear to increase across generations.

Environmental levels of atrazine decrease spatial aggregation in the freshwater mussel, Elliptio complanata.

The commonly used herbicide atrazine (ATR), may cause estrogen-like alterations and, like other xenoestrogens, may alter behavior. Here effects on animal aggregation were tested in freshwater mussels exposed short-term to ecologically relevant ATR concentrations between 1.5 and 150microg/L. The aggregation index (AI), the fraction of mussels in the most densely populated tank section, was evaluated and showed higher AI at later times in both 6- and 72-h trials (p<0.001). After 72h, there was also less aggregation in estradiol- and mid-ATR-treated animals, 26% and 24% less than control, respectively (p<0.04). These data suggest that freshwater mussels tend to aggregate, that 72-h exposure to ecologically relevant ATR concentrations decreases aggregation, and that estradiol exposure has a similar effect. Given that bivalve aggregation may be related to reproduction, the results suggest that low concentrations of ATR may have ecological consequences on animal populations in this species.

Forensic analysis of bicomponent fibers using infrared chemical imaging.

The application of infrared chemical imaging to the analysis of bicomponent fibers was evaluated. Eleven nominally bicomponent fibers were examined either side-on or in cross-section. In six of the 11 samples, infrared chemical imaging was able to spatially resolve two spectroscopically distinct regions when the fibers were examined side-on. As well as yielding characteristic infrared spectra of each component, the technique also provided images that clearly illustrated the side-by-side configuration of these components in the fiber. In one case it was possible to prepare and image a cross-section of the fiber, but in general the preparation of fiber cross-sections proved very difficult. In five of the 11 samples, the infrared spectra could be used to identify the overall chemical composition of the fibers, according to a published classification scheme, but the fiber components could not be spatially resolved. Difficulties that are inherent to conventional "single-point" infrared spectroscopy, such as interference fringing and sloping baselines, particularly when analyzing acrylic type fibers, were also encountered in the infrared chemical image analysis of bicomponent fibers. A number of infrared sampling techniques were investigated to overcome these problems, and recommendations for the best sampling technique are given. Chemical imaging results were compared with those obtained using conventional fiber microscopy techniques.

Bifenthrin activates homotypic aggregation in human T-cell lines.

BACKGROUND: Here, we addressed the concern that, despite the lack of overt toxicity, exposure to low levels of the common household pyrethroid pesticide, bifenthrin, could cause harm to the immune system. To do this, we measure the effect of bifenthrin on phytohemagglutinin (PHA) activation of homotypic aggregation in human T-cell lines. MATERIAL/METHODS: The human CD4+ H9, and Jurkat cell lines and the human promonocyte U937 cell line, were exposed to varying concentrations of bifenthrin. Cell viability was determined using the AlmarBlue Toxicity Assay. Concentrations of bifenthrin which did not reduce cell viability were determined and these concentrations were tested for the effect of bifenthrin on PHA-mediated homotypic aggregation. Blocking antibodies to ICAM and LFA-1 were used to disrupt aggregation and a nonspecific IgG was used as a control. RESULTS: Bifenthrin was found to be nontoxic at concentrations ranging from 10(-4) to 10(-13) M. Bifenthrin did not inhibit PHA induced cell aggregation in all cell lines tested. However, at 10(-4) M, bifenthrin to form aggregates stimulated homotypic aggregation in the H9 and Jurkat T-cell lines. The bifenthrin-induced aggregate formation, like that seen with PHA, was blocked by treating the cells with antibodies to either LFA-1 or ICAM. CONCLUSIONS: The results here show that bifenthrin activates T-cell function by stimulating ICAM/LFA-1 mediated homotypic aggregation. This data suggests that exposure to bifenthrin, even at "acceptable" limits, can increase the risk for and frequency of inflammatory responses and diseases such as asthma.

Bifenthrin inhibits neurite outgrowth in differentiating PC12 cells.

BACKGROUND: Bifenthrin is a third generation member of the synthetic pyrethroid family of insecticides. As a new pesticide within a relatively new class of pesticides, bifenthrin is considered relatively safe. Here, we used the PC12 neuronal cell line to examine the effect of bifenthrin on the formation of neurites and the potential developmental neurotoxicity of this pesticide. MATERIAL/METHODS: PC12 cells were exposed to varying concentrations of technical grade bifenthrin or Ortho Home Defense. Cell viability was determined using the AlmarBlue Toxicity Assay. Nontoxic concentrations of these chemicals were concomitantly with nerve growth factor and neurite outgrowth was assessed. RESULTS: Ortho Home Defense preparation reduced PC12 cell viability by approximately 50% and 70% at dilutions that correlate to bifenthrin concentrations of 10(-5) M and 10(-4) M, respectively. In contrast, technical grade bifenthrin, was not toxic to PC12 cells at 10(-3) M, which was the highest concentration tested that was soluble. At "nontoxic" concentrations of 10(-7) M and 10(-6) M, the Ortho Home Defense inhibited nerve growth factor-mediated neurite outgrowth by 30% and 55% respectively. Furthermore the nontoxic concentrations of technical grade bifenthrin of 10(-6) M and 10(-3) M inhibited neurite outgrowth by approximately 35% and 75% respectively. CONCLUSIONS: These data suggest that the toxicity of the Ortho Home Defense preparation was due to the "inert" additives in the preparation and not the bifenthrin itself. Further, these data suggest that, even in the absence of overt toxicity, bifenthrin may have deleterious effects to developing nervous system.

Forensic applications of infrared chemical imaging: multi-layered paint chips.

This paper examines the potential of infrared chemical (hyperspectral) imaging as a technique for the forensic analysis of automotive paint chips in particular, and multicomponent (e.g., layered) samples in general. Improved sample preparation procedures for the infrared analysis of paint chips are detailed, with the recommendation that where mounting resins are chemically incompatible with the sample, it is better to mount and section the sample in a soft wax from which the sections can be removed and pressed into a KBr disk for transmission analysis. Infrared chemical images of multilayered paint chips have been successfully obtained, with the chief advantage over conventional infrared analysis being that thousands of infrared spectra are collected in a few minutes across the whole sample, at a spatial resolution of around 5 microm. As with conventional infrared spectroscopy, chemical species can be identified from their spectra, but the wealth of information available can be also extracted in a number of different ways that make multicomponent spectral (and hence chemical) comparisons between two samples easy to visualize and understand. In one approach, the infrared chemical images of two paint chips being compared side-by-side can be viewed as a "movie," in which each frame is an intensity map of the two samples at a given wavenumber (frequency) value. In another approach, the spectra (pixels) in the image files are classified into chemically similar groups, resulting in a "cluster" image that makes it possible to simultaneously compare all of the layers in two paint chips. These methods are applicable to other multicomponent samples, and also to other chemical imaging techniques.

Evaluation of iodine-benzoflavone and ruthenium tetroxide spray reagents for the detection of latent fingermarks at the crime scene.

The performance of two spray reagents, iodine-benzoflavone and ruthenium tetroxide (RTX), was evaluated and compared with the conventional technique currently used at the crime scene, that is, powdering. Neither the spray techniques nor powdering were shown to be suitable for all surfaces and ages of marks tested. On some surfaces such as glass and treated wood, powdering was still the superior technique, whereas the spray techniques produced better development on wallpaper, vinyl, and brick. Sequencing work showed that RTX was incompatible with powdering and cyanoacrylate (with a rhodamine 6G stain). Iodine-benzoflavone can be used successfully either before or after powdering in a sequence; however, it was incompatible with cyanoacrylate. Two non-CFC formulations of iodine-benzoflavone using HFC4310mee and HFE7100 solvents were tested and shown not to be as effective as the original Arklone (CFC-113) formulation; however, the HFC4310mee solvent is recommended as the most suitable replacement solvent. Due to the expense of the commercial RTX spray, attempts at formulating a more cost-effective version were also carried out. A formulation was developed that gave comparable development to the commercial version but at a much cheaper cost, and with a shelf life of up to two months. Recommendations are presented for which techniques are suitable for different surfaces and ages of marks. Powdering was shown to be the best technique on all ages of marks tested on treated wood, glass, and also on marks aged three days and older on paint. Iodine-benzoflavone was the best technique on wallpaper, vinyl, brick, and raw wood. RTX was the best detection technique for fresh marks and marks aged up to one day on wallpaper and paint.