Targeting biotypes of Dactylopius tomentosus to improve effective biocontrol of Cylindropuntia spp. in Australia

Seven Dactylopius tomentosus (Lamarck) biotypes were collected from a range of Cylindropuntia spp. in Mexico, South Africa and United States of America (USA) and imported into quarantine facilities at the Ecosciences Precinct. Host range trials were conducted for each biotype and further assessed against the Cylindropuntia species that are naturalised in Australia to determine the most effective biotype for each species. Host range was confined to the Cylindropuntia for all seven biotypes. In the efficacy trials, C. imbricata (Haw.) F.M.Knuth was killed by the ‘imbricata’ biotype within 16 weeks and C. kleiniae (DC.) F.M.Knuth died within 26 weeks. Cylindropuntia fulgida var. mamillata (DC.) Backeb. and C. imbricata were killed by the ‘fulgida’ biotype within 18 weeks. On-going trials suggest that C. rosea (DC.) Backeb. could be controlled by either the ‘acanthocarpa’ or the ‘acanthocarpa × echinocarpa’ biotypes. Cylindropuntia spinosior (Englem.) F.M.Knuth was not susceptible to any of the D. tomentosus biotypes assessed. A clear designation of which D. tomentosus biotype is most suited for each Cylindropuntia species will improve and increase the effectiveness of biological control of these weed species

The host range of three biotypes of Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae) and their potential as biological control agents of Cylindropuntia spp. (Cactaceae) in Australia

The host range of two newly imported biotypes of Dactylopius tomentosus and their potential as biological control agents of Cylindropuntia spp. were investigated. A third biotype (imbricata) of D. tomentosus previously released in Australia to control C. imbricata was also screened to determine if it will feed on other species of Cylindropuntia occurring in Australia. Efficacy trials were conducted to evaluate the ability of the biotypes to retard the growth or kill those plant species supporting development of four or more individuals in the host test trials. The host range of the three biotypes of D. tomentosus was restricted to the genus Cylindropuntia. However, the biotypes showed varying degrees of specificity within this genus. The imbricata biotype was the only biotype to develop on Australian C. rosea provenances, albeit with a range of developmental success on all C. rosea provenances tested. The Spanish provenance supported the highest development success followed by Grawin (NSW), Lorne Station (NSW) while the least preferred was the Mexican provenance. The rosea and cholla biotypes were unsuitable candidates to control C. rosea in Australia. However, the efficacy trials showed that the cholla biotype had a high impact on four of the eight naturalised Cylindropuntia species in Australia. This biotype established rapidly and the sustained feeding of one fecund female and her progeny killed potted plants of C. imbricata and C. fulgida at week 18. This biotype has the potential to be an effective agent against C. fulgida, C. imbricata, C. kleiniae and C. tunicata and, as a consequence, an application seeking its release in Australia has been lodged.

Otolith morphology of four mackerel species (Scomberomorus spp.) in Australia: Species differentiation and prediction for fisheries monitoring and assessment

Four species of large mackerels (Scomberomorus spp.) co-occur in the waters off northern Australia and are important to fisheries in the region. State fisheries agencies monitor these species for fisheries assessment; however, data inaccuracies may exist due to difficulties with identification of these closely related species, particularly when specimens are incomplete from fish processing. This study examined the efficacy of using otolith morphometrics to differentiate and predict among the four mackerel species off northeastern Australia. Seven otolith measurements and five shape indices were recorded from 555 mackerel specimens. Multivariate modelling including linear discriminant analysis (LDA) and support vector machines, successfully differentiated among the four species based on otolith morphometrics. Cross validation determined a predictive accuracy of at least 96% for both models. An optimum predictive model for the four mackerel species was an LDA model that included fork length, feret length, feret width, perimeter, area, roundness, form factor and rectangularity as explanatory variables. This analysis may improve the accuracy of fisheries monitoring, the estimates based on this monitoring (i.e. mortality rate) and the overall management of mackerel species in Australia.

Ecological impacts of Phlebiopsis gigantea biocontrol treatment against Heterobasidion spp. as revealed by fungal community profiling and population analyses

Wood decay fungi belonging to the species complex Heterobasidion annosum sensu lato are among the most common and economically important species causing root rot and stem decay in conifers of the northern temperate regions. New infections by these pathogens can be suppressed by tree stump treatments using chemical or biological control agents. In Finland, the corticiaceous fungus Phlebiopsis gigantea has been formulated into a commercial biocontrol agent called Rotstop (Verdera Ltd.). This thesis addresses the ecological impacts of Rotstop biocontrol treatment on the mycoflora of conifer stumps. Locally, fungal communities within Rotstop-treated and untreated stumps were analyzed using a novel method based on DGGE profiling of small subunit ribosomal DNA fragments amplified directly from wood samples. Population analyses for P. gigantea and H. annosum s.l. were conducted to evaluate possible risks associated with local and/or global distribution of the Rotstop strain. Based on molecular community profiling by DGGE, we detected a few individual wood-inhabiting fungal species (OTUs) that seemed to have suffered or benefited from the Rotstop biocontrol treatment. The DGGE analyses also revealed fungal diversity not retrieved by cultivation and some fungal sequence types untypical for decomposing conifer wood. However, statistical analysis of DGGE community profiles obtained from Rotstop-treated and untreated conifer stumps revealed that the Rotstop treatment had not caused a statistically significant reduction in the species diversity of wood-inhabiting fungi within our experimental forest plots. Locally, ISSR genotyping of cultured P. gigantea strains showed that the Rotstop biocontrol strain was capable of surviving up to six years within treated Norway spruce stumps, while in Scots pine stumps it was sooner replaced by successor fungal species. In addition, the spread of resident P. gigantea strains into Rotstop-treated forest stands seemed effective in preventing the formation of genetically monomorphic populations in the short run. On a global scale, we detected a considerable level of genetic differentiation between the interfertile European and North American populations of P. gigantea. These results strongly suggest that local biocontrol strains should be used in order to prevent global spread of P. gigantea and hybrid formation between geographically isolated populations. The population analysis for H. annosum s.l. revealed a collection of Chinese fungal strains that showed a high degree of laboratory fertility with three different allopatric H. annosum s.l. taxa. However, based on the molecular markers, the Chinese strains could be clearly affiliated with the H. parviporum taxonomical cluster, which thus appears to have a continuous distribution range from Europe through southern Siberia to northern China. Keywords: Rotstop, wood decay, DGGE, ISSR fingerprinting, ribosomal DNA

CsrA interacting small RNAs in Haemophilus spp genomes: a theoretical analysis

The csrA is a carbon storage regulator gene that encodes a protein with multiple RNA interaction sites. Bacterial non-coding small RNAs like csrB, csrC and their counterparts in diverse bacterial genus are identified to control the regulatory activities of CsrA and its orthologs. An attempt has been made in this study to identify 'novel' non-coding small RNAs that are involved in the regulatory activities of csrA gene. All CsrA-interacting small RNAs are computationally fingerprinted to have multiple occurrence of 7-nucleotide CsrA interacting repeats [CAGGA(U/A/C)G] along with a 18-nucleotide upstream binding site. However, in several of the genomes like Haemophilus spp, the upstream binding site is not identified. The current methodology overcomes this difficulty by identifying small RNA-specific orphan transcriptional units within the intergenic regions of the genome. The results could identify all known CsrA-interacting small RNAs in E. coli, Vibrio cholerae and Pseudomonas aeruginosa genomes and additionally has picked six new possible CsrA-interacting small RNA regions in E. coli. Our computational analysis indicates that known rygD and rprA sRNAs in E. coli could possibly interact with CsrA proteins. The study is extended to three of the Haemophilus genomes that could identify seven new possible CsrA interacting small RNAs.

Finnish cattle as reservoir of Campylobacter spp.

The reported incidence of human campylobacteriosis in Finland is higher than in most other European countries. A high annual percentage of sporadic infections is of foreign origin, although a notable proportion of summer infections is domestically acquired. While chickens appear to be a major source of campylobacters for humans in most countries, the prevalence of campylobacters is very low in chicken slaughter batches in Finland. Data on other potential animal reservoirs of human pathogenic campylobacters in Finland are scarce. Consequently, this study aimed to investigate the status of Finnish cattle as a potential source of thermophilic Campylobacter spp. and antibiotic-resistant Campylobacter jejuni for human sporadic campylobacter infections of domestic origin. A survey of the prevalence of thermophilic Campylobacter spp. in Finnish cattle studied bovine rectal faecal samples (n=952) and carcass surface samples (n=948) from twelve Finnish slaughterhouses from January to December 2003. The total prevalence of Campylobacter spp. in faecal samples was 31.1%, and in carcass samples 3.5%. Campylobacter jejuni, the most common species, was present in 19.5% of faecal samples and in 3.1% of carcasses. In addition to thermophilic Campylobacter spp., C. hyointestinalis ssp. hyointestinalis was present in bovine samples. The prevalence of campylobacters was higher among beef cattle than among dairy cattle. Using the enrichment method, the number of positive faecal samples was 7.5 times higher than that obtained by direct plating. The predominant serotypes of faecal C. jejuni, determined by serotyping with a set of 25 commercial antisera for heat-stable antigens (Penner), were Pen2 and Pen4-complex, which covered 52% of the samples. Genotyping with pulsed-field gel electrophoresis (PFGE) using SmaI restriction yielded a high diversity of C. jejuni subtypes in cattle. Determining the minimum inhibitory concentrations of ampicillin, enrofloxacin, erythromycin, gentamicin, nalidixic acid, and oxytetracycline among bovine C. jejuni isolates using a commercial broth microdilution method yielded 9% of isolates resistant to at least one of the antimicrobials examined. No multiresistant isolates were found among the bovine C. jejuni strains. The study of the shedding patterns of Campylobacter spp. among three Finnish dairy cattle herds included the examination of fresh faecal samples and tank milk samples taken five times, as well as samples from drinking troughs taken once during the one-year study. The semiquantitative enrichment method detected C. jejuni in 169 of the 340 faecal samples, mostly at low levels. In addition, C. jejuni was present in one drinking trough sample. The prevalence between herds and sampling occasions varied widely. PFGE, using SmaI as restriction enzyme, identified only a few subtypes in each herd. In two 2 of the herds, two subtypes persisted throughout the sampling. Individual animals presented various shedding patterns during the study. Comparison of C. jejuni isolates from humans, chickens and cattle included the design of primers for four new genetic markers selected from completely sequenced C. jejuni genomes 81-176, RM1221 and NCTC 11168, and the PCR examination of domestic human isolates from southern Finland in 1996, 2002 and 2003 (n=309), chicken isolates from 2003, 2006 and 2007 (n=205), and bovine isolates from 2003 (n=131). The results revealed that bovine isolates differed significantly from human and chicken isolates. In particular, the - glutamyl transpeptidase gene was uncommon among bovine isolates. The PFGE genotyping of C. jejuni isolates, using SmaI and KpnI restriction enzymes, included a geographically representative collection of isolates from domestic sporadic human infections, chicken slaughter batches, and cattle faeces and carcasses during the seasonal peak of campylobacteriosis in the summer of 2003. The study determined that 55.4% of human isolates were indistinguishable from those of chickens and cattle. Temporal association between isolates from humans and chickens was possible in 31.4% of human infections. Approximately 19% of the human infections may have been associated with cattle. However, isolates from bovine carcasses and human cases represented different PFGE subtypes. In conclusion, this study suggests that Finnish cattle is a notable reservoir of C. jejuni, the most important Campylobacter sp. in human enteric infections. Although the concentration of these organisms in bovine faeces appeared to be low, excretion can be persistent. The genetic diversity and presence or absence of marker genes support previous suggestions of host-adapted C. jejuni strains, and may indicate variations in virulence between strains from different hosts. In addition to chickens, Finnish cattle appeared to be an important reservoir and possible source of C. jejuni in domestic sporadic human infections. However, sources of campylobacters may differ between rural and urban areas in Finland, and in general, the transmission of C. jejuni of bovine origin probably occurs via other routes than food.

Cultivation of Pleurotus spp. on a combination of anaerobically digested plant material and various agro-residues

Two species of Pleurotus, Pleurotus florida and Pleurotus flabellatus were cultivated on two agro-residues (paddy straw; PS and coir pith; CP) singly as well as in combination with biogas digester residue (BDR, main feed leaf biomass). The biological efficiency, nutritional value, composition and nutrient balance (C, N and P) achieved with these substrates were studied. The most suitable substrate that produced higher yields and biological efficiency was PS mixed with BDR followed by coir pith with BDR. Addition of BDR with agro-residues could increase mushroom yield by 20-30%. The biological efficiency achieved was high for PS + BDR (231.93% for P. florida and 209.92% for P. flabellatus) and for CP + BDR (14831% for P. florida and 188.46% for P. flabellatus). The OC (organic carbon), TKN (nitrogen) and TP (phosphate) removal of the Pleurotus spp. under investigation suggests that PS with BDR is the best substrate for growing mushroom. (C) 2015 Published by Elsevier Inc. on behalf of International Energy Initiative.

Copper oxide nanoparticles: an antidermatophytic agent for Trichophyton spp.

Copper oxide (CuO) is one of the most important transition metal oxides due to its unique properties. It is used in various technological applications such as high critical temperature, superconductors, gas sensors, in photoconductive applications and so on. Recently, it has been used as an antimicrobial agent against various pathogenic bacteria. In the present investigation, we studied the structural and antidermatophytic properties of CuO nanoparticles (NPs) synthesized by a precipitation technique. Copper sulfate was used as a precursor and sodium hydroxide as a reducing agent. Scanning electron microscopy (SEM) showed flower-shaped CuO NPs and X-ray diffraction (XRD) pattern showed the crystalline nature of CuO NPs. These NPs were evaluated against two prevalent species of dermatophytes, i.e. Trichophyton rubrum and T. mentagrophytes by using the broth microdilution technique. Further, the NPs activity was also compared with synthetic sertaconazole. Although better antidermatophytic activity was exhibited with sertaconazole as compared to NPs, being synthetic, sertaconazole may not be preferred, as it shows different adverse effects. Trichophyton mentagrophytes is more susceptible to NPs than T. rubrum. A phylogenetic approach was applied for predicting differences in susceptibility of pathogens.

Evaluación de tres densidades de siembra de Clitoria spp. en la producción de biomasa forrajera en base a materia seca

El presente trabajo preliminar tuvo como objetivo evaluar, el efecto de tres densidades de siembra, sobre la producción de biomasa forrajera en base a materia seca de Clitoria spp. bajo riego en época seca y su valor económico. El estudio se llevó a cabo en la Hacienda "Santa Rosa", ubicada al norte de la comunidad de Sabana Grande, Municipio de Managua. Se utilizó un diseño de bloques completos al azar (BCA), en un arreglo sencillo de tres densidades de siembra, con 42, 63 y 84 kg/ha. Se realizaron dos cortes; el primero, a los 57 días después de su establecimiento y, el segundo, 57 días después de realizado el primer corte. Los resultados mostraron que, las densidades de siembra sobre la producción de biomasa en base a materia seca total por hectárea (PBMSTH}, para cada momento de corte realizado, reflejó su mayor valor en la densidad de 84 kg/ha, con 3.153 y 4.433 ton/MS/há para el 1 10 y 2do corte, sin embargo, estos no tuvieron diferencias significativas (P>0.05) sobre las demás densidades (42 y 63 kg/ha), respectivamente. Por otra parte, las variables altura (ALT) y cobertura (COB) mostraron su mayor valor para la densidad (84 kg/ha), con 59.87 cm y 93.33 o/o respectivamente, pero estos no presentaron diferencias significativas (P>0.05) sobre las demás densidades. La prueba de rangos múltiples de Tukey, para los efectos de densidad de siembra sobre la PBMSTH, ALT y COB, no presentaron diferencias significativas. (P> 0.05; P> 0.01). De todas las densidades evaluadas, los menores costos de producción de forraje fueron para la densidad 42 kg/ha con C$ 4127.00.

Comportamiento agronómico de variedades forrajeras vignas spp. Finca Santa Rosa, Managua, Nicaragua

Se llevó a cabo un estudio con el objetivo de evaluar el comportamiento agronómico de especies y variedades de Vignas forrajeras (Vigna unguiculata, Vigna unguiculata, Vigna unguiculata, Vigna unguiculata y Vigna radiata). El Ensayo se realizó durante el período Agosto – Diciembre, 2009.en la Finca Santa Rosa, Universidad Nacional Agraria. Managua, Nicaragua. Los tratamiento consistieron en cuatro variedades de Vigna unguiculata de reciente introducción y un testigo de Vigna radiata.. El diseño utilizado fue un BCA (Bloques completo al Azar) con tres repeticiones. Las variables de estudio para cada tratamiento fueron, porcentajes de germinación, Altura de plantas (cm), Número de hojas (Conteo visual), Porcentaje de cobertura. Se realizaron análisis de varianza (ANDEVA) y separaciones de medias, usando Duncan (P<0.05). Para el análisis estadístico las variables codificadas en porcentajes se transformaron, según, arco seno 2 p (Arco seno de dos veces la raíz cuadrada de la proporción). Los resultados encontrados demuestran diferencias significativas (P ≤ 0.05) para todas las variables de estudio. El porcentaje de germinación varío de 61 % a 82 % para Vigna unguiculata CV Vun 1 y Vigna radiata CV Mungo respectivamente, mientras la Altura de plantas vario de 17.31 cm. a 33.32 cm. para Vigna unguiculata CV Vun 2 y V.unguiculata CV Vun 1. El mayor número de hojas se presentó en Vigna unguiculata CV Vun Naranja y el menor lo presentó Vigna radita CV Mungo con 10 y 4 hojas respectivamente. La mayor cobertura del suelo se presentó en Vigna unguiculata CV Vun Naranja con 42.17 % y la menor en Vigna unguiculata CV Vun 2 con 18.48 %. Se concluye que la especie Vigna unguiculata y los cultivares Vun Naranja y Vun 1 son las que presentaron un mejor comportamiento agronómico y pueden pasar a otras etapas de evaluación para su utilización en la alimentación animal.

Validación del umbral de acción para el manejo del gusano del fruto Helicoverpa spp (Boddie) Lepidoptera: noctuidae: en el cultivo del tomate Tisma, Nicaragua

Con el objetivo de validar un método determinado por ICTA/CATIE Guatemala, utilizando un umbral de acción para el manejo de Helicoverpa spp también conocido como gusano del fruto, en el cultivo del tomate se estableció un experimento de campo en la época de riego durante el período comprendido (Diciembre 1991 - Abril 1992) en Tisma, Masaya. El estudio se realizó con la colaboración de dos productores de tomate de la zona, en la finca de los productores se establecieron dos parcelas de validación, bajo dos sistemas de manejo: 1. Los productores realizaron aplicaciones periódicas semanales utilizando insecticidas químicos. 2. Aplicación de la mezcla de DIPEL (Bacillus thurinqiensis: 800 gr/mz) y LANNATE (Metomil: 75 gr/mz) según el umbral de acción, cuando se encontró cuatro huevos y/o larvas, o un 8% de frutos dañados por Helicoverpa spp en treinta plantas. El análisis económico de los tratamientos permite afirmar que las parcelas manejadas con umbral son más rentables, se obtiene mayores ingresos y menos costos a las parcelas manejadas por productor. Bajo las condiciones de este ensayo es factible económicamente, la implementación del umbral de acción, y la aplicación de la mezcla, para el combate de Helicoverpa spp en el cultivo del tomate.