Frugivoria de larvas de Neosilba McAlpine (Diptera, Lonchaeidae) sobre Psittacanthus plagiophyllus Eichler (Santalales, Loranthaceae) no sudoeste de Mato Grosso do Sul, Brasil

Loranthaceae são plantas hemiparasitas com distribuição geográfica mundial e representadas no Brasil por seis gêneros. Os mais importantes são Phthirusa, Psittacanthus e Struthanthus, os quais parasitam uma grande diversidade de plantas hospedeiras. Este trabalho avaliou a occurrência e a flutuação sazonal de moscas infestando os frutos de Psittacanthus plagiophyllus nos municípios de Anastácio, Aquidauana e Miranda, Mato Grosso do Sul, Brasil, onde ervas-de-passarinho são disseminadas. As coletas foram realizadas de junho de 1998 a julho de 2000 para obtenção de frutos maduros de P. plagiophyllus, seus insetos associados e catalogação dos seus hospedeiros. Os insetos foram criados para identificação, sendo obtidos 1.522 adultos de Neosilba spp., destes, 612 machos foram identificados a nível específico: Neosilba bifida Strikis & Prado (6 indivíduos), N. certa (Walker) (26 indivíduos), N. pendula (Bezzi) (16 indivíduos), N. zadolicha McAlpine & Steyskal (4 indivíduos) e duas diferentes espécies: morfotipo MSP1 (478 indivíduos) e morfotipo 4 (82 indivíduos). O período de mais alta infestação por Neosilba spp. ocorreu durante agosto de 1998 e de 1999 e, a espécie morfotipo MSP1 foi significativamente mais abundante que todas as outras. A morfotipo 4 foi a segunda mais abundante, diferindo significativamente de Neosilba zadolicha. Neosilba foi o único gênero de moscas frugívoras (Tephritoidea) infestante dos frutos de P. plagiophyllus e comportou-se como um invasor primário neste hospedeiro.

FRUGIVORY OF NEOSILBA SPECIES (DIPTERA: LONCHAEIDAE) AND THEPYTUS ECHELTA (LEPIDOPTERA: LYCAENIDAE) ON PSITTACANTHUS (SANTALALES: LORANTHACEAE) IN ECOTONAL CERRADO-SOUTH PANTANAL, BRAZIL

This paper presents a survey of the insects that feed on fruits of Psittacanthus Martius (Santalales: Loranthaceae), a hemiparasitic mistletoe genus that infects trees in Brazil and other neotropical countries. The aim of the study was to identify candidate insects for biological control of Psittacanthus mistletoes. Unripe and mature fruits were collected in several localities of Cerrado, bordering South Pantanal, Southwestern Brazil, from 29 Apr 1998 to 30 Jul 2000. A total of 24,710 fruits (54 samples) of Psittacanthus acinarius infecting 15 species from 10 plant families were evaluated. Psittacanthus acinarius (Mart.) was the most abundant and frequent species of mistletoe parasitizing trees in the ecotonal Cerrado-Pantanal. From 24,710 fruits of Psittacanthus acinarius were obtained 1,812 insect larvae including 1,806 Neosilba McAlpine (Diptera: Lonchaeidae) species and 6 Thepytus echelta (Hewitson) (Lepidoptera: Lycaenidae). From these emerged 1,550 Neosilba spp. adults and 6 T. echelta. Neosilba pantanense Strikis was described from this research. Larvae of T. echelta occurred in fruits of P. acinarius parasitizing Cecropia pachystachya Trecul (Urticaceae) and Anadenanthera colubrina (Vellozo) Brenan (Fabaceae). Larvae of Neosilba caused no adverse effects on the germination of infected fruits of Psittacanthus, because they do not eat the embryo or viscin tissues. This differs from the larvae of T. echelta that interrupted the germination of seeds by feeding on those tissues. Thepytus echelta may be a promising insect for the biological control of P. acinarius in the ecotonal Cerrado-Pantanal, although its abundance and frequency were low throughout the sampling period.

Larval endoparasitoids (Hymenoptera) of frugivorous flies (Diptera, Tephritoidea) reared from fruits of the cerrado of the State of Mato Grosso do Sul , Brazil

This paper presents a five years survey of endoparasitoids obtained from the larvae of frugivorous Tephritidae and Lonchaeidae flies. The insects were reared from cultivated and wild fruits collected in areas of the cerrado in the State of Mato Grosso do Sul, Brazil. The flies obtained from 14 host fruit species were eight Anastrepha species, Ceratitis capitata (Wiedemann, 1824) (Tephritidae); Dasiops sp. and Neosilba spp. (Lonchaeidae). Eleven parasitoid species were collected: Braconidae - Asobara anastrephae (Muesebek, 1958), Doryctobracon areolatus (Szépligeti, 1911), D. fluminensis (Costa Lima, 1938), Opius bellus Gahan, 1930 and Utetes anastrephae (Viereck, 1913); Figitidae - Aganaspis nordlanderi Wharton, 1998, Lopheucoila anastrephae (Rhower, 1919), Odontosema anastrephae (Borgmeier, 1935) and Trybliographa infuscata Gallardo, Díaz & Uchôa-Fernandes, 2000 and, Pteromalidae - Spalangia gemina Boucek, 1963 and S. endius Walker, 1839. In all cases only one parasitoid emerged per puparium. D. areolatus was the most abundant and frequent parasitoid of fruit fly species, as was L. anastrephae in Neosilba spp. larvae. This is the first record of A. nordlanderi in the midwestern Brazilian region.

Dinâmica populacional e incidência de moscas-das-frutas e parasitoides em cultivares de pessegueiros (Prunus Persica L. Batsch) no município de presidente prudente-sp

Esta pesquisa teve como objetivos avaliar a dinâmica populacional e registrar a diversidade de moscas-das-frutas (Diptera: Tephritoidea) em cultivares de pessegueiro Tropical, Talismã, Aurora 2, Aurora 1, Dourado 2 e Doçura 2, enxertadas sobre os porta-enxertos 'Okinawa' e Umê, em Presidente Prudente-SP. Foram realizadas as correlações da dinâmica populacional com a temperatura e a precipitação, e também a infestação com as características químicas dos frutos, Sólidos Solúveis e Acidez Titulável. No período de julho de 2004 a dezembro de 2006, a dinâmica populacional de moscas-das-frutas foi obtida através de coletas semanais de moscas-das-frutas em armadilhas McPhail, e a incidência foi determinada através da coleta de 30 frutos/planta/cultivar. O delineamento estatístico adotado foi o inteiramente casualizado, com cinco repetições. Ceratitis capitata foi predominante nas cultivares de pessegueiros estudadas. Não foi observada correlação significativa entre população de moscas-das-frutas e as variáveis de temperatura e precipitação, e sólidos solúveis e ácidez titulável. Entre as cultivares de pêssego, Aurora 2 apresentou maior infestação por C. capitata, da ordem de 22 e 23% nos anos 2004 e 2006, respectivamente. Também foi registrada a incidência de Neosilba spp. em frutos de pêssego. Doryctobracon areolatus (Braconidae), Tetrastichus giffardianus (Eulophidae) e Pachycrepoideus vindemmiae (Pteromalidae) foram recuperados de pupários de Tephritidae.

Dinâmica populacional e incidência de moscas-das-frutas e parasitoides em cultivares de pessegueiros (Prunus Persica L. Batsch) no município de presidente prudente-sp

Esta pesquisa teve como objetivos avaliar a dinâmica populacional e registrar a diversidade de moscas-das-frutas (Diptera: Tephritoidea) em cultivares de pessegueiro Tropical, Talismã, Aurora 2, Aurora 1, Dourado 2 e Doçura 2, enxertadas sobre os porta-enxertos 'Okinawa' e Umê, em Presidente Prudente-SP. Foram realizadas as correlações da dinâmica populacional com a temperatura e a precipitação, e também a infestação com as características químicas dos frutos, Sólidos Solúveis e Acidez Titulável. No período de julho de 2004 a dezembro de 2006, a dinâmica populacional de moscas-das-frutas foi obtida através de coletas semanais de moscas-das-frutas em armadilhas McPhail, e a incidência foi determinada através da coleta de 30 frutos/planta/cultivar. O delineamento estatístico adotado foi o inteiramente casualizado, com cinco repetições. Ceratitis capitata foi predominante nas cultivares de pessegueiros estudadas. Não foi observada correlação significativa entre população de moscas-das-frutas e as variáveis de temperatura e precipitação, e sólidos solúveis e ácidez titulável. Entre as cultivares de pêssego, Aurora 2 apresentou maior infestação por C. capitata, da ordem de 22 e 23% nos anos 2004 e 2006, respectivamente. Também foi registrada a incidência de Neosilba spp. em frutos de pêssego. Doryctobracon areolatus (Braconidae), Tetrastichus giffardianus (Eulophidae) e Pachycrepoideus vindemmiae (Pteromalidae) foram recuperados de pupários de Tephritidae.

Infestación natural de Lonchaeidae (Diptera) en variedades de melocotón

Se estudió la ocurrencia de Lonchaeidae en variedades de melocotón, conducidos sobre los patrones ‘Okinawa’ y Umê: Tropical, Ouromel 3, Jóia 4, Régis, Talismã, Aurora 2, Aurora 1, Dourado 2 y Doçura 2. Se colectaron muestras de 30 frutos por planta en el Banco de Germoplasma en Presidente Prudente, Brasil. Se obtuvieron 633 especímenes de Lonchaeidae; 394 de ellos en frutos de variedades sobre ‘Okinawa’ y 239 sobre Umê. Se capturaron especies como Neosilba zadolicha, N. inesperata, N. pendula, N. certa y Neosilba spp. (hembras). N. zadolicha y N. inesperata, se observaron en 77,78% de las muestras de las variedades sobre ‘Okinawa’. Sobre el patrón Umê la mayor incidencia fue de N. zadolicha , N. inesperata y N. pendula (55,6%, 33,3% y 33,3% respectivamente). Las variedades Ouromel 3, Talismã, Doçura 2 y Aurora 2 presentaron mayores infestaciones por Lonchaeidae. N. certa tuvo una menor incidencia y sólo se observó en la variedad Doçura 2 sobre ‘Okinawa’. Plantas con mayor número de frutos presentaron mayor incidencia de moscas por fruto; sin embargo, no hubo correlación entre peso del fruto y número de moscas. No se observó diferencia para peso del fruto y número de moscas por fruto entre los dos patrones, ‘Okinawa’ y Umê. Melocotones crecidos en los patrones ‘Okinawa’ y Umê en Presidente Prudente están infestados por especies de Lonchaeidae que presentan preferencia independientemente del peso de los frutos. El aporte de este trabajo al conocimiento de la comunidad de especies de Lonchaeidae ofrece una base para su control.

Moscas-Das-Frutas em Cultivares de Cafeeiros de Presidente Prudente, SP

Coffee is considered the primary host of the Mediterranean fruit fly Ceratitis capitata (Wiedemann). The infestation of C. capitata in the State of São Paulo has reached economic importance in coffee plantations. Although little information about changes caused by the fly on the coffee beverage, it is known that the fruit fly infestation causes rapid change from cherry to raisin stage, causing qualitative damage on the parchment coffee production. The objective of this work was to study the population dynamics and diversity of Tephritidae and Lonchaeidae in coffee cultivars of Icatu Vermelho IAC 4045, Vermelho IAC 99, Novo Mundo 388-17-1, Obatã IAC 1669-20, Icatu Amarelo IAC 2944, grafted on Apoatã (IAC 2258) (Coffea canephora) and Icatu Vermelho IAC 4045 ungrafted and Apoatã (IAC 2258)-grafted with approximately 3 years. The experiment was conducted in Presidente Prudente, São Paulo State, Brazil, from June 2006 to July 2008. The experimental design was a randomized block with four replications in a factorial design (3 years X 6 cultivars). From each plot of 100 plants we randomly collected 250 mature fruits. The samples were taken at 15 day intervals. The population dynamics were evaluated by using one plastic McPhail trap per cultivar. After 26 months a total of 36,932 specimens of C. capitata were trapped in all cultivars, corresponding to 49.27% males and 50.73% females. Approximately 83.3% of the specimens were collected from January to December 2007. The population fluctuation showed population peaks in May, June and July, relative to fruit ripening period. We trapped 21 specimens of A. montei Lima and A. fraterculus (Wied.). The coffee fruits of Presidente Prudente, SP, are infested by the following species of Lonchaeidae: Neosilba pendula (Bezzi), N. zadolicha McAlpine & Steyskal, N. inesperata Strikis & Prado and Neosilba pradoi Strikis & Lerena. Neosilba pendula occurred in all evaluated cultivars and N. inesperata was recoverd only from Icatu Amarelo IAC 2944 and IAC Icatu Vermelho 4045/un-grafted. No Anastrepha specimen was recovered from the fruits. Cultivars did not differ due to tephritid and lonchaeid infestations, but in 2008 the highest infestation by C. capitata occurred in the field.

Identification Of Corynebacterium Spp. Isolated From Bovine Intramammary Infections By Matrix-assisted Laser Desorption Ionization-time Of Flight Mass Spectrometry.

Corynebacterium species (spp.) are among the most frequently isolated pathogens associated with subclinical mastitis in dairy cows. However, simple, fast, and reliable methods for the identification of species of the genus Corynebacterium are not currently available. This study aimed to evaluate the usefulness of matrix-assisted laser desorption ionization/mass spectrometry (MALDI-TOF MS) for identifying Corynebacterium spp. isolated from the mammary glands of dairy cows. Corynebacterium spp. were isolated from milk samples via microbiological culture (n=180) and were analyzed by MALDI-TOF MS and 16S rRNA gene sequencing. Using MALDI-TOF MS methodology, 161 Corynebacterium spp. isolates (89.4%) were correctly identified at the species level, whereas 12 isolates (6.7%) were identified at the genus level. Most isolates that were identified at the species level with 16 S rRNA gene sequencing were identified as Corynebacterium bovis (n=156; 86.7%) were also identified as C. bovis with MALDI-TOF MS. Five Corynebacterium spp. isolates (2.8%) were not correctly identified at the species level with MALDI-TOF MS and 2 isolates (1.1%) were considered unidentified because despite having MALDI-TOF MS scores >2, only the genus level was correctly identified. Therefore, MALDI-TOF MS could serve as an alternative method for species-level diagnoses of bovine intramammary infections caused by Corynebacterium spp.

Behavioral Changes Caused By Austrodiplostomum Spp. In Hoplias Malabaricus From The São Francisco River, Brazil.

Traira (Hoplias malabaricus) is a neotropical fish that is widely distributed in freshwater environments in South America. In the present study, we documented the occurrence of metacercariae of Austrodiplostomum spp. (Diplostomidae) in the eyes and cranial cavity of H. malabaricus and described parasite-induced behavioral changes in the host. The fish were collected from the upper São Francisco River, in the Serra da Canastra mountain range, Minas Gerais, transported alive to the laboratory, observed for 2 weeks, and subsequently examined for parasites. Of the 35 fish examined, 28 (80 %) had free metacercariae in the vitreous humor (mean intensity=95.4; mean abundance=76.3), and 24 (68.57 %) had free metacercariae in the cranial cavity, mainly concentrated below the floor of the brain, at the height of the ophthalmic lobe (mean intensity=12.91; mean abundance=8.85). Specimens of H. malabaricus with a high intensity of infection in the brain displayed changes in swimming behavior.

Molecular Genetic Variability Of Commercial And Wild Accessions Of Passion Fruit (passiflora Spp.) Targeting Ex Situ Conservation And Breeding.

Passiflora species are distributed throughout Latin America, and Brazil and Colombia serve as the centers of diversity for this genus. We performed cross-species amplification to evaluate 109 microsatellite loci in 14 Passiflora species and estimated the diversity and genetic structure of Passiflora cincinnata, Passiflora setaceae and Passiflora edulis. A total of 127 accessions, including 85 accessions of P. edulis, a commercial species, and 42 accessions of 13 wild species, were examined. The cross-species amplification was effective for obtaining microsatellite loci (average cross-amplification of 70%). The average number of alleles per locus (five) was relatively low, and the average diversity ranged from 0.52 in P. cincinnata to 0.32 in P. setacea. The Bayesian analyses indicated that the P. cincinnata and P. setacea accessions were distributed into two groups, and the P. edulis accessions were distributed into five groups. Private alleles were identified, and suggestions for core collections are presented. Further collections are necessary, and the information generated may be useful for breeding and conservation.

Bartonella Spp. Bacteremia In Blood Donors From Campinas, Brazil.

Bartonella species are blood-borne, re-emerging organisms, capable of causing prolonged infection with diverse disease manifestations, from asymptomatic bacteremia to chronic debilitating disease and death. This pathogen can survive for over a month in stored blood. However, its prevalence among blood donors is unknown, and screening of blood supplies for this pathogen is not routinely performed. We investigated Bartonella spp. prevalence in 500 blood donors from Campinas, Brazil, based on a cross-sectional design. Blood samples were inoculated into an enrichment liquid growth medium and sub-inoculated onto blood agar. Liquid culture samples and Gram-negative isolates were tested using a genus specific ITS PCR with amplicons sequenced for species identification. Bartonella henselae and Bartonella quintana antibodies were assayed by indirect immunofluorescence. B. henselae was isolated from six donors (1.2%). Sixteen donors (3.2%) were Bartonella-PCR positive after culture in liquid or on solid media, with 15 donors infected with B. henselae and one donor infected with Bartonella clarridgeiae. Antibodies against B. henselae or B. quintana were found in 16% and 32% of 500 blood donors, respectively. Serology was not associated with infection, with only three of 16 Bartonella-infected subjects seropositive for B. henselae or B. quintana. Bartonella DNA was present in the bloodstream of approximately one out of 30 donors from a major blood bank in South America. Negative serology does not rule out Bartonella spp. infection in healthy subjects. Using a combination of liquid and solid cultures, PCR, and DNA sequencing, this study documents for the first time that Bartonella spp. bacteremia occurs in asymptomatic blood donors. Our findings support further evaluation of Bartonella spp. transmission which can occur through blood transfusions.

Prevalence Of Treponema Spp. In Endodontic Retreatment-resistant Periapical Lesions.

This study investigated the presence of the Treponema species in longstanding endodontic retreatment-resistant lesions of teeth with apical periodontitis, the association of this species with clinical/radiographic features, and the association among the different target species. Microbial samples of apical lesions were collected from twenty-five adult patients referred to endodontic surgery after unsuccessful root canal retreatment. Nested-PCR and conventional PCR were used for Treponema detection. Twenty-three periradicular tissue samples showed detectable levels of bacterial DNA. Treponema species were detected in 28% (7/25) of the cases. The most frequently detected species were T. socranskii (6/25), followed by T. maltophilum (3/25), T. amylovorum (3/25), T. lecithinolyticum (3/25), T. denticola (3/25), T. pectinovorum (2/25) and T. medium (2/25). T. vicentii was not detected in any sample. Positive statistical association was found between T. socranskii and T. denticola, and between T. maltophilum and T. lecithinolyticum . No association was detected between the presence of any target microorganism and the clinical or radiographic features. Treponema spp. are present, in a low percentage, in longstanding apical lesions from teeth with endodontic retreatment failure.

Propriedades mecânicas de peças com dimensões estruturais de Pinus spp: correlação entre resistência à tração e classificação visual

A redução da disponibilidade de espécies de madeiras nativas e seus efeitos na economia, associada ao fortalecimento dos conceitos de preservação ambiental, criou a necessidade de desenvolvimento de alternativas viáveis para utilização racional de espécies de reflorestamento. E uma das opções é a realização de classificação visual das peças. Autores de trabalhos desenvolvidos nessa linha de pesquisa verificaram a adequação das regras de classificação visual do Southern Pine Inspection Bureau (SPIB) dos EUA à madeira de Pinus do Brasil e apresentaram proposta para normalizar o processo de classificação visual dessa madeira. Nessa classificação, os aspectos com maior influência são: presença de nós, desvio de grã em relação ao eixo da peça e densidade de anéis de crescimento. Assim, esta pesquisa apresenta um estudo experimental que consistiu na classificação visual e determinação da resistência à tração de 85 peças de Pinus spp e um estudo teórico, que propôs uma equação para determinar a resistência à tração média de peças estruturais em função da classificação visual. Com este trabalho, foi possível observar a influência dos nós e dos anéis de crescimento sobre a resistência à tração das peças analisadas.

Leishmania spp. parasite isolation through inoculation of patient biopsy macerates in interferon gamma knockout mice

Isolation of Leishmania parasite and species identification are important for confirmation and to help define the epidemiology of the leishmaniasis. Mice are often used to isolate pathogens, but the most common mouse strains are resistant to infection with parasites from the Leishmania (Viannia) subgenus. In this study we tested the inoculation of interferon gamma knockout (IFNγ KO) mice with biopsy macerates from Leishmania-infected patients to increase the possibility of isolating parasites. Biopsies from twenty five patients with clinical signs of leishmaniasis were taken and tested for the presence of parasites. Immunohistochemical assay (IHC) and conventional histopathology detected the parasite in 88% and 83% of the patients, respectively. Leishmania sp. were isolated in biopsy macerates from 52% of the patients by culture in Grace's insect medium, but 13% of isolates were lost due to contamination. Inoculation of macerates in IFNγ KO mice provides isolation of parasites in 31.8% of the biopsies. Most isolates belong to L. (Viannia) subgenus, as confirmed by PCR, except one that belongs to L. (Leishmania) subgenus. Our preliminary results support the use of IFNγ KO mice to improve the possibility to isolate New World Leishmania species.

Protocol for DNA extraction of Cryptosporidium spp. oocysts in fecal samples

Molecular characterization of Cryptosporidium spp.oocysts in clinical samples is useful for public health since it allows the study of sources of contamination as well as the transmission in different geographical regions. Although widely used in developed countries, in Brazil it is restricted to academic studies, mostly using commercial kits for the extraction of genomic DNA, or in collaboration with external reference centers, rendering the method expensive and limited. The study proposes the application of the modifications recently introduced in the method improving feasibility with lower cost. This method was efficient for clinical samples preserved at -20 °C for up to six years and the low number of oocysts may be overcomed by repetitions of extraction.