Distribution and prevalence of the entomopathogenic nematode Steinernema feltiae in Scotland

The distribution of insect parasitic nematodes in Scotland was investigated between April 1988 and April 1989 using the wax moth, Galleria mellonella, larvae as bait. Of the 1014 samples examined only 2.2% were found to be positive and all the nematodes recovered and identified belonged to the one species Steinernema feltiae. A more detailed investigation into the distribution of S. feltiae in a 1 ha block indicated that it had a very aggregated distribution.     

Field test of the nematodeSteinernema feltiae (Nematoda: Steinernematidae) against yellowjacket colonies (Hym.: Vespidae)

The insect parasitic nematodeSteinernema carpocapsae (Weiser) was applied to 4 colonies ofVespula wasps in the field. At all treated colonies worker activity was reduced by at least 50% after one week. Two treated colonies partially recovered, and 2 colonies were completely destroyed.Vespula workers removed nematodekilled and dying nestmates from treated colonies. The results suggest that under certain conditions, nematodes could be used as biological control agents of yellowjackets.      

Measuring abundance,diversity and parasitic ability in two populations of the nematophagous fungus Pochonia chlamydosporia var. chlamydosporia

Abundance, genetic diversity and parasitic ability in the facultative nematode parasite Pochonia chlamydosporia var. chlamydosporia were compared in soils from two sites in Portugal under long-term tomato cultivation where root-knot nematodes (Meloidogyne sp.) were present. Fungal abundance assessed by selective agar or real-time quantitative PCR with specific primers was similar in both soils. PCR fingerprinting of isolates with ERIC primers indicated that the dominant P. c. var. chlamydosporia biotypes (profiles A and B) in both soils were very closely related, although a second biotype (profile C) was detected in one soil. When tomato plants infected with M. incognita were grown in the two soils, only profiles A and B were recovered from eggs. Primers based on polymorphisms in vcp1 demonstrated that isolates with profiles A and B were likely to prefer root-knot nematodes, whereas profile C preferred cyst nematodes. In the soil containing profiles A, B and C, egg parasitism by P. chlamydosporia was estimated at 1% using water agar plates with antibiotics but fewer than 0.2% of M. incognita eggs were shown to be infected with P. c. var. chlamydosporia when using species-specific β-tubulin-PCR primers. In contrast, the soil containing only profile B showed 22% egg parasitism on water agar plates and more than 2.5% of eggs were confirmed as P. c. var. chlamydosporia by species-specific β-tubulin-PCR primers. The results, which reveal limited diversity within the fungus at the two sites, are discussed in relation to biological control of plant-parasitic nematodes.     

Molecular profiling of nematode associates with Rhynchophorus ferrugineus in southern Italy

A survey of nematodes associated with the red palm weevil Rhynchophorus ferrugineus was conducted in southern Italy in 2015 and 2016 in order to create a species inventory and obtain data about nematode biodiversity. A total of 70 insect samples (pupae and adults) were collected from infested Phoenix canariensis, Phoenix dactylifera, and Chamaerops humilis palms in three Italian Regions: sampling took place at 11 locations in Apulia, 1 in Basilicata, and 1 in Sardinia regions. Individual insects were dissected to determine nematode presence, and different nematode species were also recovered from red palm weevil cocoons collected at the sites in Apulia. Individual nematodes were molecularly identified by sequencing the ITS, D2‐D3 expansion domains of the 28SrRNA gene and the mitochondrial COI and inferring the phylogenetic relationships. The insect‐associated nematofauna identified belonged to the families Rhabditidae, Cephalobidae, and Diplogastridae. Just two nematode species, Teratorhabditis synpapillata and Mononchoides macrospiculum, were always found in association with adult insects and cocoons taken from all sampling sites. This paper reports on the biodiversity of the nematodes associated with R. ferrugineus and on current knowledge of the specific habitat of specialized and divergent entomophilic nematodes.     

Diversity and distribution of entomopathogenic nematodes (Steinernematidae, Heterorhabditidae) in South Africa

A total of 1506 soil samples from different habitats in seven geographic regions of South Africa were evaluated for the presence of entomopathogenic nematodes (EPN). Nematodes were isolated from 5% of the samples. Among the steinernematids, four Steinernema sp. were recovered including Steinernema khoisanae and three new undescribed species. Although steinernematids were recovered from both humid subtropical and semiarid regions, this family accounted for 80% of EPN recovered from the semiarid climate zones characterised by sandy, acidic soils. Eight isolates of S. khoisanae were recovered from the Western Cape province. One of the new undescribed steinernematids (Steinernema sp. 1) was recovered only from the Free State and KwaZulu-Natal provinces where humid subtropical conditions prevail and soils are generally less acidic with higher clay content. A high level of adaptation, however, was noted with Steinernema sp. 2, which was recovered from a wide range of soil conditions and habitats ranging from semiarid (Western Cape province) to humid subtropical (KwaZulu-Natal province). A third undescribed steinernematid, Steinernema sp. 3, seemed better adapted to heavier soils with more than 80% of isolates recovered from fruit orchards in the Free State province. Heterorhabditis bacteriophora was the only heterorhabditid recovered during this survey. This species was particularly prevalent in four provinces ranging from humid subtropical to semiarid regions. Isolation of EPN directly from insect cadavers included Steinernema sp. 2 and one H. bacteriophora from an unidentified white grub (Scarabaeidae) cadaver (i.e., dual infection) and H. bacteriophora from the black vine weevil, Otiorhynchus sulcatus.     

Control of Western flower thrips (Frankliniella occidentalis Pergande) pupae in compost

Western flower thrips (Frankliniella occidentalis) develops from an active larval stage through to a non-feeding, almost immobile, pre-pupal and pupal stage. This generally occurs in the compost or soil below the plant on which the larvae fed. Control of thrips at this stage in their development offers a chance of utilising pathogens or pesticides not suitable for use in an integrated control programme aimed at adult and larval stages. Trials were done with F. occidentalis pupae and pre-pupae in a soil/peat based compost using 11 pesticides, three fungal pathogens and four species of insect parasitic nematodes. The pesticides malathion and chlorpyrifos-methyl gave the most promising result with 97.5% and 96.5% control, respectively. The fungal pathogen Metarhizium anisopliae proved better when applied as a pre-pupation rather than as a post-pupation treatment (74.5% : 26.6% control). The insect parasitic nematode Steinernema carpocapsae gave 76.6% control when applied at 25 times 10⁴ nematodes litre^-1 of compost. The results are discussed in relation to control of thrips in glasshouses.     

Ecology of nematophagous fungi: distribution and habitat

A survey of 161 samples of soil and plant material collected from sites throughout Ireland has shown that nematophagous fungi exist in a wide range of habitats. 205 isolations were made consisting of 11 endoparasitic and 20 predatory species. The commonest endoparasites and their frequency of occurrence were Myzocytium spp. (9·3%), Acrostalagmus obovatus (6·8%) and Harposporium anguillulae (6·2%). Predators were equally abundant, the commonest being Dactylella bembicodes (8·7%), D. mammillata (6·8%) and D. ellipsospora (6·2%). Arthrobotrys oligospora and A. musiformis both had a highly restricted distribution. Adhesive knobs and constricting rings were significantly more abundant than the other trapping mechanisms. All the samples were classified into one of 10 broad habitat types. Of these temporary agricultural pasture, coastal vegetation and coniferous leaf litter had the greatest percentage of sites from which nematophagous fungi were isolated. A number of species showed distinct habitat associations, in particular Arthrobotrys robusta, A. musiformis and Dactylella cionopaga.     

Prevalence and antimicrobial resistance of Enterococcus species of food animal origin from Beijing and Shandong Province,China

Aims

To evaluate the prevalence and antimicrobial resistance of Enterococcus species from chickens and pigs in Beijing and Shandong Province, China.

Methods and Results

Swab samples were collected from four farms in Beijing and two in Shandong Province in 2009 and tested for Enterococcus. Minimum inhibitory concentrations of antimicrobial agents were determined using broth microdilution or agar screening methods. A total of 453 Enterococcus isolates were recovered, belonging to six different Enterococcus species. All isolates were sensitive to vancomycin. Resistance to tetracycline (92·5%), amikacin (89·4%), erythromycin (72·8%) and rifampin (58·1%), and high‐level streptomycin resistance (HLSR, 50·3%) were prevalent, while resistance to penicillins (7·9% to penicillin and 4·2% to ampicillin) was rare. The resistance rates to phenicols (chloramphenicol and florfenicol) and enrofloxacin, and high‐level gentamicin resistance (HLGR) were approximately 30%. The vast majority of the Enterococcus isolates were classified as multidrug‐resistant organisms.

Conclusions

Resistance of Enterococcus sp. to most antimicrobials was more prevalent in China than in European or other Asian countries.

Significance and Impact of the study

Our findings reveal a high level of antimicrobial resistance in Enterococcus isolates from food animals in China and underline the need for prudent use of antibiotics in chicken and pig production to minimize the spread of antibiotic‐resistant enterococci.     

Atlantic sturgeon Acipenser oxyrinchus surfacing behaviour

Atlantic sturgeon Acipenser oxyrinchus surfacing behaviour was investigated in Minas Basin (45° 20′ N; 64° 00′ W) and the Bay of Fundy with pop‐up satellite archival tags (MiniPAT) measuring physical variables (pressure, temperature, light ) . Of six tags deployed during June and July, five provided pop‐up locations and two were recovered after c. 4 months. Analysis of recovered archival data revealed that the frequency of surfacing events was highest (78·9%) when A. oxyrinchus were in Minas Basin at depths <10 m. Surfacing frequency decreased substantially when fish migrated into greater depths of the Bay of Fundy (>40 m). The tidal cycle in Minas Basin had a significant relationship to surfacing frequency, with the most surfacing events (49·5%) occurring on the flood tide, from mid‐ to high‐tide. Surfacing events ranged from 0–12 a day and the maximum number occurred between 2300 and 0300 hours. Maximum surfacing ascent speeds ranged from 0·50 to 4·17 m s^?1 and maximum descent speeds ranged from 0·17 to 3·17 m s^?1. Buoyancy control, by gulping air to inflate the gas bladder, is proposed as the main reason for surfacing behaviour in A. oxyrinchus.     

Entomopathogenic nematodes,a potential microbial biopesticide: mass production and commercialisation status – a mini review

Parasitic nematodes have several important attributes that make them excellent candidates for biological control of soil insects. These nematodes can be produced by in vivo by baiting technique on insects and commercially by in vitro solid/liquid culturing. Numerous insect pests on many different crops are being controlled by these insect parasitic nematodes, including root weevils, flea beetles, mint root borer, colorado potato beetle, white grubs, caterpillars and plant parasitic root nematode, e.g. root-knot nematodes. Utilisation of entomopathogenic nematodes (EPN) has raised intense interest and has been a growing concern globally mainly because of its potential efficiency, exemption from registration and other impressive attributes for utilising against the control of soil dwelling pests. This review highlights the mass production, commercialisation and utilisation of EPN as microbial biopesticide in bio-intensive pest management programmes.     

In vitro assays on the susceptibility of four species of nematophagous fungi to anthelmintics and chemical fungicides/antifungal drug

Using nematophagous fungi for the biological control of animal parasitic nematodes will become one of the most promising strategies in the search for alternative chemical drugs. The purpose of this study was to check the in vitro activity of four anthelmintics, four chemical fungicides and two antifungal drugs on the spore germination of nematophagous fungi: Duddingtonia flagrans (SF170), Arthrobotrys oligospora (447), Arthrobotrys superba (435) and Arthrobotrys sp. (PS011). A modified 24-well cell culture plate assay was conducted to evaluate the susceptibility of nematophagous fungi against drugs tested by calculating the effective middle concentrations (EC₅₀) of each tested drug to inhibit the germination of fungal spores. EC₅₀ ranged between 0·7 and 47·2 μg ml⁻¹ for fenbendazole, thiabendazole and ivermectin, except levamisole (546·5–4057·8 μg ml⁻¹). EC₅₀ of tested fungicides was 0·6–2·3 μg ml⁻¹ for carbendazim, 55·9–247·4 μg ml⁻¹ for metalaxyl, 24·4–45·2 μg ml⁻¹ for difenoconazole, and 555·9–1438·3 μg ml⁻¹ for pentachloronitrobenzene (PCNB). EC₅₀ of two antifungal drugs was 0·03–3·4 μg ml⁻¹ for amphotericin B and 0·3–10·9 μg ml⁻¹ for ketoconazole. The results showed that 10 tested drugs, except for levamisole and PCNB, had in vitro inhibitory effects on nematophagous fungi. The chlamydospores of D. flagrans had the highest sensitivity to nine tested drugs, except for ketoconazole.     

Incidence and survival of non-O157 verocytotoxigenic Escherichia coli in soil

Aims: This study estimated the incidence of non‐O157 verocytotoxigenic Escherichia coli (VTEC) in farm pasture soils and investigated the survival of non‐O157 VTEC in clay and sandy loam soils. Methods and Results: Twenty farms were tested over a 12‐month period by sample enrichment in tryptone soya broth plus vancomycin, followed by PCR screening for the presence of vt1 and vt2 genes. Of the 600 soil samples, 162 (27%), across all farms, were found to contain vt1 and/or vt2 genes. The enrichment cultures from the 162 PCR‐positive samples were plated onto Chromocult tryptone bile X‐glucuronide agar (TBX), presumptive VTEC colonies recovered, confirmed as VTEC by PCR and serotyped. Samples of the two predominant soil types in Ireland (clay and sandy) were homogenized, characterized in terms of pH, boron, cobalt, copper, potassium, magnesium, manganese, phosphorus, zinc and organic matter content, inoculated with washed suspensions of eight non‐O157:H7 soil isolates and six bovine faecal isolates and stored at 10°C for up to 201 days. Inoculum survival rates were determined at regular intervals by recovering and plating soil samples on TBX. All inoculated non‐O157 serotypes had highest D‐values in the sandy loam soil with D‐values ranging from 50·26 to 75·60 days. The corresponding range in clay loam soils was 31·60–48·25 days. Conclusions: This study shows that non‐O157 VTEC occur widely and frequently in pasture soils and can persist in such environments for several months, with considerable opportunity for recycling through farm environments, and cattle, with clear potential for subsequent transmission into the human food chain. Significance and Impact of the Study: This is the first such study of non‐O157 VTEC in farm soils and found that these VTEC are frequent and persistent contaminants in farm soils. In light of recent epidemiological data, non‐O157 VTEC should be seen as an emerging risk to be controlled within the food chain.     

The Effects of Soil Salinity and Meloidogyne javanica on Tomato

A non-sodic, non-saline sandy loam soil was salinized to anion-cation ratios similar to those naturally occurring in Iraq and California. The interactions of saline soils (conductivities 4, 8, 12 and 16 mmhos/cm) with a moderately salt-tolerant plant (Lycopersicon esculentum ''Marimond'') and a plant parasitic nematode (Meloidogyne javanica) were investigated. Plant parasitic nematodes were shown to be an important modifying influence within the plant environment, either accentuating or ameliorating salinity stress effects.     

Pathogenicity,reproduction and foraging behaviours of some entomopathogenic nematodes on a new turf pest,Dorcadion pseudopreissi (Coleoptera: Cerambycidae)

Entomopathogenic nematodes (EPNs) in the families Heterorhabditidae and Steinernematidae have considerable potential as biological control agents of soil-inhabiting insect pests. In the present study, the control potential of the EPNs Steinernema carpocapsae (TUR-S4), S. feltiae (Nemaplus), S. carpocapsae (Nemastar), S. feltiae (TUR-S3) and Heterorhabditis bacteriophora (Nematop) against a new longicorn pest, Dorcadion pseudopreissi Breuning, 1962 (Coleoptera: Cerambycidae), on turf was examined in laboratory studies. Pathogenicity tests were performed at the following doses: 50, 100 and 150 Dauer Juveniles (DJs)/larva at 25°C. Highest mortalities (75–92%) of the larvae were detected at the dose of 150 DJs/larva for all nematodes used. Reproduction capabilities of the used EPNs were examined at doses of 50, 75, 100 and 150 DJs/larva at 25°C. S. carpocapsae (TUR-S4) had the most invasions (32 DJs/larva) and reproduction (28042 DJs/larva) at the dose of 100 DJs, and the highest reproduction (per invaded DJ into a larva) was observed in H. bacteriophora (Nematop) (2402.85 DJs) at a dose of 50 DJs. The foraging behaviour of the nematodes in the presence of D. pseudopreissi and Galleria mellonella L. (Lepidoptera: Galleriidae) larvae was studied using a Petri dish filled with sand at 20°C. All of the used nematodes accumulated near the larvae section of both insect species (32–53% of recovered DJs) with a higher percentage of S. carpocapsae (TUR-S4) (53%) and H. bacteriophora (48%) (Nematop) moving towards larvae of D. pseudopreissi, than the S. feltiae strains.     

Nematodes associated with peas and beans in Scotland

Six hundred and sixty-four soil samples from farms growing peas and beans in eastern Scotland were examined for plant parasitic and predatory nematodes. Neither pea cyst-nematode, Heterodera goettingiana nor pea early-browning virus disease were found. Stem nematode, Ditylenchus dipsaci was recorded in broad beans once. The most frequently occurring plant parasitic nematodes were Tylenchorhynchus dubius, Longidorus elongatus, Rotylenchus goodeyi and Helicotylenchus vulgaris but numbers were generally too small to cause damage. It is concluded that although potentially damaging plant parasitic nematodes occur in Scotland, nematode-related diseases occur infrequently and are of little significance to the crop.     

Microbial contamination of food products consumed by infants and babies in Korea

Aims: The objectives of this study were to investigate the microbiological safety of various foods intended for consumption by infants and babies. Methods and Results: The incidence of Cronobacter spp. and Enterobacteriaceae from powdered infant formula (PIF, n = 75) and baby soy milk (n = 10) was examined. Additionally, aerobic plate count, coliforms and the prevalence of foodborne pathogens were investigated in 230 samples from a variety of infant and baby foods, including cereal‐based follow‐up formulas (FUF), liquid FUF and other infant foods. High APCs were observed in nutrient supplements and cereal‐based FUF. Coliforms were found in 6 (2·6%) products, and Cronobacter spp. was isolated in 10 (4·4%) samples, including four PIF and six cereal‐based FUF. Bacillus cereus was detected in 48 (20·9%) samples: cereal‐based FUF items (23·0%), rice soups (20·6%), honey samples (40·0%), biscuits (40·0%) and liquid FUF (7·4%). Conclusions: New safety criteria, along with hygienic control measures and consumer education strategies, are essential to improve the microbiological safety of infant or baby foods. Significance and Impact of the Study: This study provides comprehensive information about the prevalence and level of contamination of infant and baby food products by Cronobacter spp. and other major foodborne pathogens.     

Dispersal and Infectivity of the Entomogenous Nematode,Neoaplectana carpocapsae Weiser (Rhabditida: Steinernematidae), in Sand

Laboratory tests determined the lateral and vertical dispersal patterns of Neoaplectana carpocapsae in sand. In the vertical tests, placement of infective juveniles 15 cm below the sand''s surface resulted in the majority (77%) being recovered above the point of placement after 48 h. Placement of the nematodes on the sand''s surface resulted in the majority (90.4%) remaining within 1 cm of the sand''s surface. Placement of nematodes at depths of 2.5 cm and 5.0 cm below the sand''s surface also resulted in little nematode dispersal. However, vertical hioassay tests showed that juvenile nematodes placed on the sand''s surface dispersed 12 cm down to infect 67% of the Galleria mellonella pupae placed at the depth. Conversely, when nematodes were placed 11 cm below the insect pupae no infection was observed, but 53% infection occurred when nematodes were 7 cm below the site of the insect pupae. In lateral dispersal, 87% of the nematodes rentained within 2 cm of the placement site, although 0.5% were recovered at 12-14 cm away from the point of placement. Lateral bioassay tests indicated that the nematodes were capable of infecting 90, 35, and 5% of the G. mellonella pupae at 7 cm, 10 cm, and 14 cm from the point of placement, respectively.     

Analysis of antibiotic resistance and virulence patterns in Klebsiella pneumoniae isolated from human urinary tract infections in India

In this study, 504 urine samples collected from patients suffering from urinary tract infection (UTIs) were screened for the presence of Klebsiella pneumoniae. The overall occurrence of Klebsiella spp. and K. pneumoniae was found to be 23·2% (117/504) and 16·8% (85/504) respectively. Antibiotic susceptibility testing of 85 K. pneumoniae isolates was carried out by disc diffusion which revealed alarming levels of antibiotic resistance (ABR). Antimicrobial resistance was prominently observed against cefpodoxime (76·47%) followed by ampicillin (70·59%), ceftriaxone (52·94%), cefoxitin (50·59%), amoxyclav (48·24%), ofloxacin (45·88%), cefotaxime (44·71%), cefepime (43·53%) and doxycycline hydrochloride (40%). A small percentage of strains also exhibited resistance to other antimicrobials in the range of 7–35%. Around 77·6% of the isolates were found to be resistant to three or more antibiotic classes and 66·7% of the isolates had multiple antibiotic resistance index values >0·2. Screening of virulence genes in 85 K. pneumoniae isolates revealed that uge gene was the most predominant (11/85, 12·9%), followed by rmpA (9/85, 10·5%), kfu (4/85, 4·7%) and aerobactin genes (2/85, 2·35%). Further, the overall percentage of biofilm producers were found to be 17·65% (15/85). This study warrants hospitals and health care centres to reduce misuse of antibiotics and manage UTI with appropriate treatment after performing antibiotic susceptibility testing.     

Top 10 plant‐parasitic nematodes in molecular plant pathology

The aim of this review was to undertake a survey of researchers working with plant‐parasitic nematodes in order to determine a ‘top 10’ list of these pathogens based on scientific and economic importance. Any such list will not be definitive as economic importance will vary depending on the region of the world in which a researcher is based. However, care was taken to include researchers from as many parts of the world as possible when carrying out the survey. The top 10 list emerging from the survey is composed of: (1) root‐knot nematodes (Meloidogyne spp.); (2) cyst nematodes (Heterodera and Globodera spp.); (3) root lesion nematodes (Pratylenchus spp.); (4) the burrowing nematode Radopholus similis; (5) Ditylenchus dipsaci; (6) the pine wilt nematode Bursaphelenchus xylophilus; (7) the reniform nematode Rotylenchulus reniformis; (8) Xiphinema index (the only virus vector nematode to make the list); (9) Nacobbus aberrans; and (10) Aphelenchoides besseyi. The biology of each nematode (or nematode group) is reviewed briefly.     

Antimicrobial resistance and toxin gene profiles of Staphylococcus aureus strains from Holstein milk

Isolation of Staphylococcus aureus (Staph. aureus) from Holstein milk samples with mastitis and nonmastitis was conducted to estimate its prevalence, antimicrobial resistance and toxin genes. A total of 353 milk samples were collected from three Chinese Holstein herds. Fifty‐three Staph. aureus isolates collected from 29 Staph. aureus‐positive samples were characterized via antimicrobial susceptibility, toxin genes and Pulsed‐field Gel Electrophoresis (PFGE) profiles. The prevalence of Staph. aureus was 4·0–9·5% in mastitic and 7·3–11·5% in nonmastitic samples in the analysed herds. Approximately 61·0% of Staph. aureus strains isolated from mastitis cows were resistant to ≥10 antimicrobials compared with 0% of isolates with nonmastitis. The most frequently observed super antigenic toxin gene was pvl (41·5%) followed by seh + pvl (13·2%). We did not find mecA‐positive methicillin‐resistant Staph. aureus (MRSA) strains, while mecA‐negative MRSA strains were identified in the three herds. PFGE results suggested potential transmission of Staph. aureus strains in different farms. These results open new insights into Staph. aureus transmission and antimicrobial resistance of Holstein dairy cows and into developing strategies for udder health improvement of dairy cattle.