Selection of an ivermectin-resistant strain of Rhipicephalus microplus (Acari: Ixodidae) in Brazil

Resistance to ivermectin (IVM) in field Populations of Rhipicephalus microplus of Brazil has been observed since 2001 In this work, four selection methods (infestations with: (I) IVM-treated larvae, (2) larvae from IVM-treated adult female ticks, (3) larvae from IVM-treated adult female ticks on an IVM-treated host, and (4) larvae obtained from W-treated females that produced eggs with a high eclosion rate) were used oil a field population with an initial ivermectin (IVM) resistance ratio at LC50 (RR50) of 1 37 with the objective to obtain experimentally a highly-resistant strain After ten generations, using these methods combined, the final RR50 was 8 06 This work shows for the first time that it was possible to increase IVM resistance in R. microplus in laboratory conditions. The establishment of a drug resistant R microplus strain is a fundamental first step for further research into the mechanisms of ivermectin-resistance in R. microplus and potentially methods to control this resistance (C) 2009 Elsevier B V All rights reserved

Resistance to cypermethrin, deltamethrin and chlorpyriphos in populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from small farms of the State of Sao Paulo, Brazil

A field survey of resistance was conducted based on the larval packet test technique with synthetic pyrethroids (cypermethrin and deltamethrin) and organophosphates (chlorpyriphos) in Rhipicephalus (Boophilus) microplus field populations from six different regions of the State of Sao Paulo (Brazil). 82.6% of the populations showed resistance to cypermethrin, 86.36% to deltamethrin and 65.25% to chlorpyriphos, with 50% presenting resistance to both SP and OP acaricide. According to the questionnaires completed by the producers, OP + SP mixtures followed by SP-only formulations were the products most commonly used for controlling the cattle tick in the surveyed areas. The present study showed high occurrence of resistance to SP and OP in the State of Sao Paulo, Brazil and revealed the type of strategy adopted by small dairy farms in this state. This information is fundamental in order to establish the monitoring of resistance on each farm individually, contributing to the rational use of acaricides for the control of R. (B.) microplus. (C) 2011 Elsevier B.V. All rights reserved.

Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) resistance to fipronil in Uruguay evaluated by in vitro bioassays

Rhipicephalus (Boophilus) microplus obtained from four local populations in Uruguay (2007-2008) were subjected to various bioassay techniques to determine the presence of fipronil resistance within the country. Resistance ratios (RRs) obtained by larval immersion test varied between 3.3 and 3635 for tick populations subjected to treatment with fipronil for the last 3-7 years. The highest RR was observed in the population which received fewer treatments. Using discriminating concentration (8 ppm) for larval immersion test, all field strains were correctly diagnosed as fipronil-resistant. This study presents the first diagnoses of cattle tick resistance to fipronil in Uruguay`s field populations. It also highlights the importance of the possible conflict between programs to control agricultural pests and cattle ticks. The findings provide valuable information for selection and adoption of new control alternatives to manage drug resistance exhibited by cattle ticks. (C) 2009 Elsevier BM. All rights reserved.

In vitro tests to establish LC(50) and discriminating concentrations for fipronil against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) and their standardization

Laboratory test was carried out on larvae and adults of the cattle tick, Rhipicephalus (Boophilus) microplus, to determine fipronil toxicity. Adult immersion test (AIT, N = 26), larval immersion test (LIT, N = 71) and larval packet test (LPT, N = 41) were standardized using susceptible strain (Mozo). Dose-response curves were compared with a fipronil resistant strain. Four variables were analyzed from AIT results: mortality, weight of eggs on day 7 and on day 14, index of fertility, and index of fecundity. For larval test, dose mortality curves were analyzed. In spite of the high LC(50) variability, all variables determined for AIT were appropriate to discriminate both strains. AIT and LIT had more sensitivity than LPT, with larger resistance factors. It was used two times LC(99.9) as discriminating doses (DCs) following FAO suggestion. For mortality by AIT, LIT and LPT the DCs were estimated: 4.98 ppm, 7.64 ppm and 2365.8 ppm, respectively, for Mozo strain. DCs mortality values estimated for resistant strain by AIT, LIT and LPT were: 6.96 x 10(5) ppm, 343.26 ppm and 5.7 x 10(3) ppm, respectively and their respective resistant factors were: 202.4, 5.36 and 1.52. Protocols for AIT, LIT and LPT have been presented in this paper. (C) 2009 Elsevier B.V. All rights reserved.

Determination of the susceptibility of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) to ivermectin and fipronil by Larval Immersion Test (LIT) in Uruguay

Rhipicephalus (Boophilus) microplus is an important cattle pest in Uruguay, and the law regulates its control. It is resistant to organophosphates, synthetic pyrethroids and, as recently discovered, to fipronil. Resistance to macrocyclic lactones (MLs) and amitraz have not been documented; however, veterinarians and farmers have reported treatment failures. The objective of the present work was to study the susceptibility of cattle tick strains from different Uruguayan counties to ivermectin (IVM) and fipronil by using the Larval Immersion Test (LIT). The Mozo strain was used as the susceptible reference strain. From 2007 to 2009, twenty-eight tick populations were collected from different cattle farms with and without history of IVM or fipronil use. A probit analysis estimated dose-mortality regressions, lethal concentrations (LC), and confidence intervals. The resistance ratio (RR) was determined at the LC(50) and LC(90) estimates. To classify a tick population in relation to resistance, three categories based on a statistical analysis of LC and RR between field populations and Mozo strains were defined: susceptible (no differences), incipient resistance (differences and RR(50) < 2) and resistant (differences and RR(50) >= 2). Eighteen field populations were tested with IVM and five of them presented a RR(50) range between 1.35 and 1.98 and the LC(50/90), which is statistically different from the Mozo strain (incipient resistance). However, the RR(90) increases >= 2 in four of the populations, confirming that tick resistance to IVM is emergent. The low RR values obtained could be a result of a low frequency of treatments with IVM. Twenty-seven tick populations were tested with fipronil and six were diagnosed as resistant according to the LIT. Cross-resistance was not observed between fipronil and IVM on these tick populations. The current study presents different R. (B.) microplus populations with an incipient resistance to IVM, and indicates that the fipronil tick resistance is restricted to certain areas in Uruguay. (c) 2011 Elsevier B.V. All rights reserved.

Clonagem e caracterização parcial de cDNA de glândula salivar de Boophilus microplus (Acari:Ixodidae) similar a glutationa s-transferase

O carrapato Boophilus microplus é um ectoparasita hematófago de bovino que causa sérias perdas econômicas. Estudos para o desenvolvimento de formas alternativas de controle do carrapato tem sido realizados para diminuir ou substituir a aplicação de agentes químicos, que contaminam o ambiente, os derivados da carne, além dos problemas de resistência das gerações de carrapatos aos acaricidas. As vacinas são uma forma alternativa de controle do carrapato. As enzimas glutationa S-transferase (GSTs) são alvo potencial para intervenção imunológica contra alguns parasitas. Este trabalho teve como objetivo isolar e caracterizar parcialmente um cDNA de B. microplus similar a GST da classe Mu. O clone SG2 foi isolado dentre aproximadamente 8 x 103 pfu de fagos recombinantes de uma biblioteca de cDNA de glândula salivar de partenógina, sondada com anti-soro de coelho contra glândula salivar. O clone SG2 contendo um inserto de 864 pb teve sua seqüência determinada e a fase de leitura aberta corresponde a 220 amino ácidos. A análise desta seqüência indicou que o gene clonado codifica uma GST de B. microplus (BmGST) com um motivo altamente conservado entre os resíduos 60 e 68 que compreende o sítio de ligação a glutationa (GSH) e outro motivo SLAILRYL, centrado no resíduo 78 da seqüência. No alinhamento múltiplo da AgSG2 com outras GSTs foi observada uma similaridade de até 41% com GSTs da classe Mu de outros organismos e inclusive com outra GST da classe Mu isolada de larva de B. microplus (HE et al., 1999). A proteína recombinante AgSG2 purificada apresentou atividade enzimática contra o substrato cromogênico CDNB. Ensaios de atividade enzimática com extratos de tecidos, secreções e excreções foram realizados para verificar a presença de GST. Ensaios de RT-PCR com tecidos de B. microplus indicaram que os sítios de síntese de BmGST são glândulas salivares e intestino de partenógina e teleógina.

Some stigmaeid and eupalopsellid mites from citrus orchards in Brazil (Acari : Stigmaeidae and Eupalopsellidae)

Two new citrus-inhabiting species in the family Stigmaeidae, Agistemus brasiliensis n. sp. and Zetzellia malvinae n. sp., are described from citrus plants in São Paulo State, Brazil. Also, Agistemus floridanus Gonzalez is redescribed. Keys to genera of the Stigmaeidae and Eupalosellidae are provided along with a key to all species of these families associated with citrus.

Parâmetros biológicos e tabela de vida de Tetranychus urticae (Acari: Tetranychidae) em cultivares de mamão

Este trabalho foi realizado para avaliar o desenvolvimento e reprodução de Tetranychus urticae em cultivares comerciais de mamão Carica papaya. Foram utilizadas cultivares do grupo Formosa (Tainung 01 e Calimosa) e do grupo Solo (Golden e Sunrise). Para iniciar o bioensaio, foi transferida uma fêmea fertilizada por disco de folha (n=50 repetições) e retirado após um período de 12h, sendo avaliada a cada 12h, registrando-se o período de incubação, duração do estágio de imaturo, longevidade e fecundidade dos adultos e viabilidade desses estágios. Os resultados indicaram que as cultivares de mamão Tainung 01, Calimosa, Sunrise e Golden são bons hospedeiros para T. urticae. O parâmetro viabilidade não sofreu influência das cultivares em todas as fases de desenvolvimento avaliadas, apresentando valores superiores a 90%. Não houve diferença estatística entre as cultivares nos parâmetros: período de pre-oviposição e viabilidade dos ovos. A cultivar Tainung 01 apresentou menor potencial hospedeiro, embora houvesse menor duração nos estágios de ovo, larva, protoninfa e ovo-adulto. Nos parâmetros de tabela de vida e fertilidade, apresentou menores valores de Ro, r m e λ e maior valor de Td. Entre as cultivares, o Sunrise apresentou um elevado potencial hospedeiro para T. urticae, pois essa cultivar proporcionou a maior produção de ovos por fêmea, maior longevidade das fêmeas, bem como a maior taxa de reprodução (maior Ro, r m e λ e menor valor de Td).

A revision of the genus Pulaeus Den Heyer, with descriptions of a new genus and four new Brazilian species (Acari: Prostigmata: Cunaxidae)

A new genus, Lupaeus gen. nov., is created to contain part of the described species in the genus Pulaeus Den Heyer. A key to the genera of the tribe Pulaeini, to which these two genera belong, and one auxiliary key to the species of Brazil and South Africa, are provided. Four new species, viz. Pulaeus myrtaceus sp. nov., P. quadrisolenidius sp. nov., Lupaeus lectus sp. nov. and L. lobidorsalis sp. nov. are described and figured. P. martini Den Heyer, 1981 is designated as type species for the new genus. New nomenclatural combinations are reported.

Efeito da adição de óleos mineral e vegetal a acaricidas no controle do ácaro-da-leprose dos citros Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpide)

Com a finalidade de esclarecer os efeitos da adição de óleo vegetal e mineral aos acaricidas foi conduzido um ensaio de campo em 1994 no município de Viradouro, SP, utilizando-se de Assist, Triona e Natur'l Óleo, na dosagem de 500 mL por 100 litros de água, adicionados aos acaricidas: pyridaben nas formulações 200 CE e 750 PM, nas dosagens de 75 mL e 20 g; propargite 720 CE a 100 mL; óxido de fenbutatina 500 SC a 80 mL e cyhexatin 500 PM a 50 g. O delineamento estatístico adotado foi o de blocos casualizados. As aplicações foram feitas com pulverizador tipo pistola. Após a preparação da calda, foi determinado o pH. Empregou-se máquina de varredura e microscópio estereoscópico para as avaliações da população acarina. A adição de Natur'l Óleo pode afetar negativamente a eficiência do pyridaben 200 CE e 750 PM e cyhexatin 500 PM, no controle do ácaro-da-leprose. Triona e Assist não afetaram as eficiências dos acaricidas testados. Pelo índice de iodo, mediu-se o grau de insaturação das misturas dos acaricidas com Natur'l Óleo, concluindo-se que houve incorporação das moléculas dos acaricidas pelas ligações insaturadas do óleo; porém, isto não explica o diferente comportamento dos produtos no controle do ácaro da leprose dos citros.

Life cycle and behaviour of the predaceous mite Cunaxatricha tarsospinosa (Acari: Prostigmata: Cunaxidae)

Life cycle and behaviour of Cunaxatricha tarsospinosa Castro & Den Heyer from rubber trees in Brazil were studied, with Tenuipalpus heveae Baker offered as prey. The study was conducted at 25.4 +/- A 0.2A degrees C, 83 +/- A 5% RH and 12:12 h L:D photophase. The egg stage was the longest immature stage, lasting 17.1 +/- A 1.3 days (mean +/- A SE); total juvenile development was completed in 33.2 +/- A 2.8 days. Lifetime fecundity was 12.0 +/- A 2.2 eggs. Intrinsic rate of population increase was low, suggesting that T. heveae may not be a good prey for the predator. All specimens of C. tarsospinosa collected in the field for this study were females, no males were found. Concurrently, only females were obtained in the laboratory. This seems to be the first report of thelytokous parthenogenesis for cunaxids. Similar to earlier reports for some Cunaxinae and Coleoscirinae, prey were captured when predators were actively searching for them.

Description of a new species and complementary description of a known species of Iphiseiodes de Leon (Acari: Phytoseiidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversity of phytoseiid mites (Acari: Mesostigmata: Phytoseiidae) in the Atlantic Forest of São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

NEW PHYTOSEIIDAE (ACARI: MESOSTIGMATA) FROM COSTA RICA, WITH ADDITIONAL INFORMATION on OTHER SPECIES

This paper reports on a study of phytoseiids collected from the La Selva Biological Field Station, the ALAS (Arthropods of La Selva) Project, made available to us by the Canadian National Collection via Dr. E. E. Lindquist. This material includes two new species, Proprioseiopsis versus n. sp. and Transeius costarricensis n. sp., three new records for Central America, and additional information for nine species previously reported from La Selva.

Actinedida e Acaridida (Arachnida: Acari) edáficos de Piracicaba, estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)