Polychrome Bone Fluorescence: A valuable diagnostic tool for 4th degree calvarial burns. A Case report.

4th degree calvarial burns are complex in management with regards to determining bone vitality and subsequent choice of defect coverage. Distinguishing viable and nonviable bone can be challenging. Tetracycline bone fluorescence is well described and in common use in maxillofacial patients with bisphosphonate osteonecrosis and has been reported in septic orthopedic surgery specifically periprosthetic joint infection. On the basis that viable bone fluoresces, the concept of polychrome fluorescence was extrapolated as a diagnostic tool for calvarial burns. Bone fluorescence presents an efficient, non-invasive, and cost-effective diagnostic tool delineating exact necrotic margins in 4th degree burns ensuring targeted bone sparing debridement. This report describes the use of polychrome bone fluorescence as an intra-operative tool including a single-centre case report with 4th degree calvarial burns.

Natural infection of white grubs (Coleoptera: Scarabaeidae) with entomopathogenic nematodes in the KwaZulu-Natal province of South Africa.

White grubs are root feeding larvae of beetles (Coleoptera: Scarabaeidae) that are sporadic pests in agriculture and can lead to economic damage. The grubs feed on the roots of plants, while the adult beetle can bore into underground stems, as well as cause defoliation of plants. Sporadic incidence of larvae with symptoms of nematode infections were detected in wattle and sugarcane plantations in the KwaZulu-Natal province of South Africa. The larvae with infection symptoms were isolated, washed, and put on water traps to collect infective juveniles of possible nematode infections. Three species of entomopathogenic nematodes (EPNs) were isolated from the white grub larvae. These included Steinernema bertusi isolated from Maladera sp. 4., Oscheius myriophila from Maladera sp. 4 and Schizonchya affinis, and Steinernema fabii isolated from Maladera sp. 4., Pegylis sommeri, and S. affinis. Of these S. fabii was the most common species in the sample (87%). This is the first report of such a high diversity of locally occurring EPNs found naturally associated with white grub species in this region of South Africa.

In vitro liquid culture of the mollusc-parasitic nematode Phasmarhabditis (Rhabditida: Rhabditidae).

The success of the mollusc-parasitic nematode, Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae), as a biological control agent in Europe has led to worldwide interest in phasmarhabditids as biocontrol agents. In this study, the mass culture potential of three phasmarhabditids, namely Phasmarhabditis papillosa, Phasmarhabditis kenyaensis and Phasmarhabditis bohemica, was assessed. In addition, ten bacterial candidates, consisting of seven associated with slugs and three associated with entomopathogenic nematodes, were investigated. The bacteria were tested for their ability to cause mortality to Deroceras invadens, as well as to support nematode growth. Initial mortality studies demonstrated that Kluyvera, Aeromonas and Pseudomonas spp. (AP3) caused 100% mortality when they were injected into the haemocoel of D. invadens. However, in growth studies, Pseudomonas sp. (AP4) was found to be the most successful bacterium, leading to recovery and reproduction in almost all nematode species, except for P. kenyaensis. In flask studies, P. bohemica, which showed exceptional growth with Pseudomonas sp. (AP1), was chosen for further investigation. The effect of inoculating flasks with different concentrations of Pseudomonas sp. (AP1), as well as with different concentrations of P. bohemica, was evaluated by assessing the nematode populations for 14 days. The results indicated that the lowest, 1% (v/v), bacteria inoculation led to higher total nematode and to infective juvenile (IJ) yield, with flasks with the highest IJ inoculum (3000 IJs/ml) having a positive effect on the total number of nematodes and IJs in cultures of P. bohemica. This study presents improvements for the mass-culturing of nematodes associated with molluscs.

Palaeoclimatic changes resulted in range expansion and subsequent divergence in brown fur seals, Arctocephalus pusillus.

Past climatic change as a driving force of marine diversification is still largely unclear, particularly for Southern Hemisphere species. Here, we present a case using the brown fur seal, Arctocephalus pusillus, assessing the geographical structure and demographic history using mitochondrial and nuclear data. Results show the two previously defined subspecies (one from Australia and the other from southern Africa) are phylogeographically distinct. Migration analyses based on nuclear data suggest the absence of migrants among the two genetically close assemblages. The demographic history of A. pusillus is characterized by a glacial population expansion (approx. 18 kya) in the southern African lineage, which coincides with time estimates of population expansion of prey species of seals. Approximate Bayesian calculations support an eastward dispersal event during the Last Glacial Maximum when sea levels were lower, followed by a postglacial divergence event, approximately 13 kya. The demographic history of the brown fur seal in the Southern Oceans provides support that recent palaeoclimatic changes could have facilitated expansions in some marine species and that postglacial sea-level rise may have acted as a dispersal barrier for species mostly confined to continental shelves.

Ex vivo development of Phasmarhabditis spp. associated with terrestrial molluscs.

The success of Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae) as a biological control agent of molluscs has led to a worldwide interest in phasmarhabditids. However, scant information is available on the lifecycle development of species within the genus. In the current study, the development of P. hermaphrodita, Phasmarhabditis papillosa, Phasmarhabditis bohemica and Phasmarhabditis kenyaensis were studied using ex vivo cultures, in order to improve our understanding of their biology. Infective juveniles (IJs) of each species were added to 1 g of defrosted homogenized slug cadavers of Deroceras invadens and the development monitored after inoculated IJ recovery, over a period of eight-ten days. The results demonstrated that P. bohemica had the shortest development cycle and that it was able to produce first-generation IJs after eight days, while P. hermaphrodita, P. papillosa and P. kenyaensis took ten days to form a new cohort of IJs. However, from the perspective of mass rearing, P. hermaphrodita has an advantage over the other species in that it is capable of forming self-fertilizing hermaphrodites, whereas both males and females are required for the reproduction of P. papillosa, P. bohemica and P. kenyaensis. The results of the study contribute to the knowledge of the biology of the genus and will help to establish the in vitro liquid cultures of different species of the genus.

South African cardiovascular risk stratification guideline for non-cardiac surgery.

Executive summary The South African (SA) guidelines for cardiac patients for non-cardiac surgery were developed to address the need for cardiac risk assessment and risk stratification for elective non-cardiac surgical patients in SA, and more broadly in Africa.The guidelines were developed by updating the Canadian Cardiovascular Society Guidelines on Perioperative Cardiac Risk Assessment and Management for Patients Who Undergo Non-cardiac Surgery, with a search of literature from African countries and recent publications. The updated proposed guidelines were then evaluated in a Delphi consensus process by SA anaesthesia and vascular surgical experts. The recommendations in these guidelines are:1. We suggest that elective non-cardiac surgical patients who are 45 years and older with either a history of coronary artery disease, congestive cardiac failure, stroke or transient ischaemic attack, or vascular surgical patients 18 years or older with peripheral vascular disease require further preoperative risk stratification as their predicted 30-day major adverse cardiac event (MACE) risk exceeds 5% (conditional recommendation: moderate-quality evidence).2. We do not recommend routine non-invasive testing for cardiovascular risk stratification prior to elective non-cardiac surgery in adults (strong recommendation: low-to-moderate-quality evidence).3. We recommend that elective non-cardiac surgical patients who are 45 years and older with a history of coronary artery disease, or stroke or transient ischaemic attack, or congestive cardiac failure or vascular surgical patients 18 years or older with peripheral vascular disease should have preoperative natriuretic peptide (NP) screening (strong recommendation: high-quality evidence).4. We recommend daily postoperative troponin measurements for 48 - 72 hours for non-cardiac surgical patients who are 45 years and older with a history of coronary artery disease, or stroke or transient ischaemic attack, or congestive cardiac failure or vascular surgical patients 18 years or older with peripheral vascular disease, i.e. (i) a baseline risk >5% for MACE 30 days after elective surgery (if no preoperative NP screening), or (ii) an elevated B-type natriuretic peptide (BNP)/N-terminal-prohormone B-type natriuretic peptide (NT-proBNP) measurement before elective surgery (defined as BNP >99 pg/mL or a NT-proBNP >300 pg/mL) (conditional recommendation: moderate-quality evidence).Additional recommendations are given for the management of myocardial injury after non-cardiac surgery (MINS) and medications for comorbidities.

Occurrence and Seasonal Changes in the Population of Root-knot Nematodes on Honeybush (Cyclopia Sp.).

Root-knot nematodes in the genus Meloidogyne are an important group of plant-parasitic nematodes causing severe damage on agricultural crops worldwide. A study was conducted to identify the species of root-knot nematodes causing damage on honeybush monocultures and to assess the seasonal variations in the nematode population. Soil samples were collected from six experimental sites in Genadendal, Western Cape province of South Africa from 2016 to 2017. DNA was extracted from single-second stage juveniles and species identifi cation was done using species-specifi c sequence-characterised amplifi ed regions (SCAR) primers. Meloidogyne hapla and M. javanica were identifi ed from the sites. Mean population density of the nematodes varied significantly (p < 0.05) in the six sites, with the peak population being recorded in summer of 2017. The study suggests that seasonal variation in temperature and moisture could contribute to changes in the population density of root-knot nematodes in the soil.

Nematodes that associate with terrestrial molluscs as definitive hosts, including Phasmarhabditis hermaphrodita (Rhabditida: Rhabditidae) and its development as a biological molluscicide.

Terrestrial molluscs (Mollusca: Gastropoda) are important economic pests worldwide, causing extensive damage to a variety of crop types, and posing a health risk to both humans and wildlife. Current knowledge indicates that there are eight nematode families that associate with molluscs as definitive hosts, including Agfidae, Alaninematidae, Alloionematidae, Angiostomatidae, Cosmocercidae, Diplogastridae, Mermithidae and Rhabditidae. To date, Phasmarhabditis hermaphrodita (Schneider, 1859) Andrássy, 1983 (Rhabditida: Rhabditidae) is the only nematode that has been developed as a biological molluscicide. The nematode, which was commercially released in 1994 by MicroBio Ltd, Littlehampton, UK (formally Becker Underwood, now BASF) under the tradename Nemaslug®, is now sold in 15 different European countries. This paper reviews nematodes isolated from molluscs, with specially detailed information on the life cycle, host range, commercialization, natural distribution, mass production and field application of P. hermaphrodita.

First Report of the Isolation of the Symbiotic Bacterium Photorhabdus luminescens subsp. laumondii Associated with Heterorhabditis safricana from South Africa.

Photorhabdus luminescens subsp. laumondii is closely associated with the entomopathogenic nematode Heterorhabditis bacteriophora and has, to date, not been isolated from other nematode species. This study is the first report of P. luminescens subsp. laumondii from two South African isolates of entomopathogenic nematodes, Heterorhabditis safricana SF281 and H. bacteriophora SF351. Both symbiotic bacterial strains are phenotypically closely related to P. luminescens subsp. laumondii previously isolated and described from H. bacteriophora. The genetic relatedness between P. luminescens subsp. laumondii strains SF281B and SF351B was confirmed by comparing 16S rDNA, recA, gyrB and gltX sequences with sequences of P. luminescens subsp. laumondii, including the type strain (TT01T) and strain E21.

Entomopathogenic nematodes for the control of the codling moth (Cydia pomonella L.) in field and laboratory trials.

Three commercially available entomopathogenic nematode (EPN) strains (Steinernema feltiae and Heterorhabditis bacteriophora Hb1 and Hb2) and two local species (S. jeffreyense and S. yirgalemense) were evaluated for the control of the codling moth (Cydia pomonella). In field spray trials, the use of S. jeffreyense resulted in the most effective control (67%), followed by H. bacteriophora (Hb1) (42%) and S. yirgalemense (41%). Laboratory bioassays using spray application in simulated field conditions indicate S. feltiae to be the most virulent (67%), followed by S. yirgalemense (58%). A laboratory comparison of the infection and penetration rate of the different strains showed that, at 14°C, all EPN strains resulted in slower codling moth mortality than they did at 25°C. After 48 h, 98% mortality was recorded for all species involved. However, the washed codling moth larvae, cool-treated (at 14°C) with S. feltiae or S. yirgalemense, resulted in 100% mortality 24 h later at room temperature, whereas codling moth larvae treated with the two H. bacteriophora strains resulted in 68% and 54% control, respectively. At 14°C, S. feltiae had the highest average penetration rate of 20 IJs/larva, followed by S. yirgalemense, with 14 IJs/larva. At 25°C, S. yirgalemense had the highest penetration rate, with 39 IJs/larva, followed by S. feltiae, with 9 IJs/larva. This study highlights the biocontrol potential of S. jeffreyense, as well as confirming that S. feltiae is a cold-active nematode, whereas the other three EPN isolates tested prefer warmer temperatures.

Steinernema jeffreyense n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from South Africa.

During a non-targeted survey for entomopathogenic nematodes in South Africa, a new species of Steinernema was isolated from a soil sample collected from underneath a guava tree, close to the shore at Jeffrey's Bay. The nematode was isolated by means of the insect-baiting technique using last-instar larvae of Galleria mellonella. It is described herein as Steinernema jeffreyense n. sp. The nematode can be separated from other described, closely related species in terms of the morphological and morphometric characteristics of the different life stages, and in terms of the characterization and phylogeny of DNA sequences of the internal transcribed spacer (ITS) rDNA of the 18S gene, and of the D2D3 region of the 28S rDNA gene. The new species is placed molecularly in the arenarium-glaseri-karii-longicaudatum group characterized by the following morphological characters: infective third-stage juvenile with a body length of 926 (784-1043) µm, distance from head to excretory pore of 87 (78-107) µm, tail length of 81 (50-96) µm, with an E% of 109 (86-169), and eight evenly spaced ridges (i.e. nine lines) in the middle of the body. First-generation males have a spicule length of 88 (79-95) µm and gubernaculum length of 57 (51-61) µm. Male mucron is absent in both generations. First-generation females have an asymmetrical protuberance and a short, double-flapped epiptygmata, with both flaps directed to the front. The tail of the first-generation female is shorter than the anal body width, with a mucron on the dorsal tail tip, with D% = 78 (59-99). Cross-hybridization with S. khoisanae, S. tophus and S. innovationi showed the new species to isolate reproductively from the others. The analyses of ITS rDNA and D2D3 sequence of the 18S and 28S rDNA genes support the studied nematode isolate to be a valid new species belonging to the 'glaseri' group (Clade V).

Development and population dynamics of Steinernema yirgalemense (Rhabditida: Steinernematidae) and growth characteristics of its associated Xenorhabdus indica symbiont in liquid culture.

Entomopathogenic nematodes have become a valuable addition to the range of biological control agents available for insect control. An endemic nematode, Steinernema yirgalemense, has been found to be effective against a wide range of key insect pests. The next step would be the mass production this nematode for commercial application. This requires the establishment of monoxenic cultures of both the nematode and the symbiotic bacterium Xenorhabdus indica. First-stage juveniles of S. yirgalemense were obtained from eggs, while X. indica was isolated from nematode-infected wax moth larvae. The population density of the various life stages of S. yirgalemense during the developmental phase in liquid culture was determined. The recovery of infective juveniles (IJs) to the third-stage feeding juveniles, was 67 ± 10%, reaching a maximum population density of 75,000 IJs ml- 1 on day 13 after inoculation. Adult density increased after 8 days, with the maximum female density being 4600 ml- 1 on day 15, whereas the maximum male density was 4300 ml- 1 on day 12. Growth curves for X. indica showed that the exponential phase was reached 15 h after inoculation to the liquid medium. The stationary phase was reached after 42 h, with an average of 51 × 107 colony-forming units ml- 1. Virulence tests showed a significant difference in insect mortality between in vitro- and in vivo-produced nematodes. The success obtained with the production of S. yirgalemense in liquid culture can serve as the first step in the optimizing and upscaling of the commercial production of nematodes in fermenters.

First report of the symbiotic bacterium Xenorhabdus indica associated with the entomopathogenic nematode Steinernema yirgalemense.

The entomopathogenic nematode Steinernema yirgalemense is considered a promising agent in the biocontrol of insects. However, little is known about the bacteria living in symbiosis with the nematode. In this study, we have identified the only available bacterial strain (157-C) isolated from S. yirgalemense, as a member of the species Xenorhabdus indica. Identification was based on 16S rDNA, recA, dnaN, gltX, gyrB and infB gene sequence analyses. The relatedness of strain 157-C to the type strain of X. indica (DSM 17 382) was confirmed with DNA-DNA hybridization. The phenotypic characteristics of strain 157-C are similar to those described for the type strain of X. indica. This is the first report associating X. indica with S. yirgalemense.

Adjuvants to improve aerial control of the citrus mealybug Planococcus citri (Hemiptera: Pseudococcidae) using entomopathogenic nematodes.

The citrus mealybug, Planococcus citri, is a highly destructive pest of citrus, occurring only in the aerial parts of plants. Humidity will be one of the key factors to consider when using entomopathogenic nematodes (EPN) as biological control agents. Different adjuvants can be added to suspensions of EPNs, to improve control as a foliar application. An aqueous suspension containing Heterorhabditis zealandica and 0.3% Zeba® significantly increased P. citri mortality by 22% at 80% relative humidity (RH) with a temperature cycle starting at 22°C for 14 h and 11°C for 11 h. The same polymer formulation was tested for Steinernema yirgalemense and mortality of P. citri increased by 21% at 60% RH and by 27% at 80% RH. The addition of Nu-Film-P® and Zeba® to H. zealandica suspensions did not significantly retard application run-off on citrus leaves. The combination of Nu-Film-P® and Zeba®, however, was able to significantly retard sedimentation, increasing the average number of nematodes deposited on 2-cm2 leaf discs by 10 nematodes. In an aqueous suspension, nematodes settle rapidly to the bottom, resulting in an uneven distribution of nematodes. Xanthan gum, at a concentration of 0.2%, was highly effective at retarding sedimentation, with 72% of the initial nematode number still in suspension after 1 h. Zeba®, at a concentration of 0.3%, despite not being as effective as Xanthan gum, nevertheless still retarded sedimentation significantly. This is the first report of the potential of Nu-Film-P® and Zeba® to improve EPN performance against P. citri when used above ground in citrus orchards.

Photovoice: a novel approach to improving antituberculosis treatment adherence in pune, India.

We compared antituberculosis treatment (ATT) adherence and outcomes among patients exposed to Photovoice (video of previously cured TB patients sharing experiences about TB treatment) versus those not exposed. The odds of successful outcome (i.e., cured or completing treatment) for the 135 patients who watched Photovoice were 3 times greater (odds ratio: 2.8; 95% CI: 1.3-6.1) than for patients who did not watch Photovoice. The comparison group, on average, missed more doses (10.9 doses; 95% CI: 6.6-11.1) than the intervention group who saw Photovoice (5.5 doses; 95% CI: 3.7-6.1). Using Photovoice at initiation of ATT has the potential to improve treatment adherence and outcomes.

Potential of entomopathogenic nematodes for the control of the banded fruit weevil, Phlyctinus callosus (Schönherr) (Coleoptera: Curculionidae).

Entomopathogenic nematodes (EPN) were evaluated for their potential use as biological control agents against Phlyctinus callosus, the banded fruit weevil (BFW). The susceptibility of larvae and adults to EPN was evaluated using 400 infective juveniles (IJ) per insect after 4 days in 24-well bioassay trays. The nematode isolates used were all able to infect BFW, although the larvae were found to be more susceptible than were the adults. The percentage mortality for BFW larvae ranged from 41 to 73% and for BFW adults from 13 to 45%. The most effective isolate, SF41 of Heterorhabditis zealandica, was used to investigate the effect of vertical movement of nematodes in sand and sandy loam soil, at specified concentration and temperature. A higher (82.2 ± 0.084%) percentage mortality rate was obtained with the sandy loam soil, than with the use of sand (67.5 ± 0.12%). The LD50 and LD90 values after 4 days of incubation were 96 and 278 IJ/50 µl, respectively. Nematodes were inactive below 15 °C, with the highest mortality of 74 ± 0.081% for BFW larvae recorded at 25 °C. Heterorhabditis zealandica was able to complete its life cycle successfully in sixth-instar BFW larvae after a period of 22 days. The study showed BFW larvae not to be as susceptible to nematode infection as they need a high concentration (400 IJ/larva) and 4 days to give effective control.

Heterorhabditis noenieputensis n. sp. (Rhabditida: Heterorhabditidae), a new entomopathogenic nematode from South Africa.

A new entomopathogenic nematode in the genus Heterorhabditis is described from South Africa, from two singular isolates found 1000 km from each other, from beneath a fig tree and in a citrus orchard, respectively. Morphological and molecular studies indicate both isolates to be the same and a new undescribed Heterorhabditis species. Comparison of sequences of the internal transcribed spacer (ITS) rDNA and the D2D3 region of the 28S rDNA gene with available sequences of other described species within the genus, indicate the two isolates as a new species. Phylogenetic analysis of the sequence data concerned placed the new species, H. noenieputensis n. sp., closest to H. indica and H. gerrardi in the indica-group. The new species, H. noenieputensis n. sp., is distinguished from other species in the genus by a combination of several morphological traits of the males and the infective juveniles (IJs). The new species differs from all other species previously described, as regards the body length of the IJs, except for H. indica and H. taysearae, in which the IJ is smaller. The IJ also differs from that of H. indica in the length of the oesophagus, the body diameter, the length of the tail and the E%. In addition, males of H. noenieputensis n. sp. differ from their closest relative, H. indica, in the position of the excretory pore, SW% and D%; and from H. gerrardi in the length of the oesophagus and SW%. The seventh pair of genital papillae of H. noenieputensis n. sp. are normally developed, while for H. indica they are often branched or swollen at the base, while 8 and 9 are usually absent in both species.

ORGANIC VS CONVENTIONAL: SOIL NEMATODE COMMUNITY STRUCTURE AND FUNCTION.

Global increases in human population are creating an ever-greater need for food production. Poor soil management practices have degraded soil to such an extent that rapidly improved management practices is the only way to ensure future food demands. In South Africa, deciduous fruit producers are realising the need for soil health, and for an increased understanding of the benefits of soil ecology, to ensure sustainable fruit production. This depends heavily on improved orchard management. Conventional farming relies on the addition of artificial fertilizers, and the application of chemicals, to prevent or minimise, the effects of the soil stages of pest insects, and of plant-parasitic nematodes. Currently, there is resistance toward conventional farming practices, which, it is believed, diminishes biodiversity within the soil. The study aimed to establish the soil nematode community structure and function in organically, and conventionally, managed deciduous fruit orchards. This was done by determining the abundance, the diversity, and the functionality of the naturally occurring free-living, and plant-parasitic, nematodes in deciduous fruit orchards in the Western Cape province of South Africa. The objective of the study was to form the basis for the use of nematodes as future indicators of soil health in deciduous fruit orchards. Orchards from neighbouring organic, and conventional, apricot farms, and from an organic apple orchard, were studied. All the nematodes were quantified, and identified, to family level. The five nematode-classified trophic groups were found at each site, while 14 families were identified in each orchard, respectively. Herbivores were dominant in all the orchards surveyed. Organic apples had the fewest herbivores and fungivores, with the highest number of carnivores. When comparing organic with conventional apricot orchards, higher numbers of plant-parasitic nematodes were found in the organic apricot orchards. The Maturity Index (MI) indicated that all orchard soils had values below 1.5, indicating disturbed conditions. The conventionally managed apricot orchard had the highest MI value, of 1.48. The Plant Parasitic Index (PPI) value was highest in the organically managed apricot orchard. In order to determine the existing enrichment, structural, and basal conditions, the nematode faunal analysis was applied to each site. All the sites indicated enriched and structured conditions. Regarding the diversity, the richness, and the evenness of the distribution, soil from the conventional apricot orchard had the highest species richness, whereas the organic apple orchard soil had the most even distribution of families. Different management practices in fruit orchards did not show marked differences in terms of community composition and structure.