Abstract The ground beetle, Neoaulacoryssus speciosus (Coleoptera: Carabidae) is of high relevance to field because it has been recorded as a pest of seeds and young plants of vegetables and other crops and a predator in agricultural crops, forest and weeds in Brazil. However, natural habitat changes are increasing agriculture and forest insect outbreaks in urban areas. A N. speciosus population outbreak occurred in October and November 2018 simultaneously in 12 neighboring municipalities at the beginning of the rainy season in the northern region of Minas Gerais State, Brazil. The objectives of this study were to report a sudden and simultaneous population outbreak of N. speciosus and to describe the factors of habitat change that could have contributed to this invasion in 12 municipalities in the northern region of Minas Gerais State in the Caatinga biome of Brazil. In addition, female and male genitals were described and illustrated, the scientific classification revised and common names of N. speciosus listed. Thousands of males and females of N. speciosus agglomerated in shady, humid places during the day and night for about 15 days. Neoaulacoryssus speciosus has been identified and illustrated, its scientific classification revised and four common names listed for this species. beetle Coleoptera (Coleoptera Carabidae Brazil However areas N 201 1 addition illustrated shady days species 20 2

Abstract Euryades corethrus is a Troidini butterfly (Papilionidae, Papilioninae), endemic to grasslands in southern Brazil, Uruguay, Argentina and Paraguay. Formerly abundant, nowadays it is in the Red list of endangered species for those areas. During its larval stage, it feeds on Aristolochia spp, commonly found in southern grasslands. These native grassland areas are diminishing, being converted to crops and pastures, causing habitat loss for Aristolochia and E. corethrus. This study aimed to assess the genetic diversity, population structure and demographic history of E. corethrus. We sampled eight populations from Rio Grande do Sul, Brazil and based on Cytochrome Oxidase subunit I (COI) molecular marker, our results suggest a low genetic variability between populations, presence of gene flow and, consequently, lack of population structure. A single maternally inherited-genetic marker is insufficient for population-level decisions, but barcoding is a useful tool during early stages of population investigation, bringing out genomic diversity patterns within the target species. Those populations likely faced a bottleneck followed by a rapid expansion during the last glaciation and subsequent stabilization in effective population size. Habitat loss is a threat, which might cause isolation, loss of genetic variability and, ultimately, extinction of E. corethrus if no habitat conservation policy is adopted. Papilionidae, Papilionidae (Papilionidae Papilioninae, Papilioninae , Papilioninae) Uruguay Paraguay abundant stage spp diminishing pastures E Sul COI (COI consequently inheritedgenetic inherited populationlevel level decisions investigation size threat isolation ultimately adopted

Resumo A Leishmaniose Visceral Canina (LVC) causada pela Leishmania (Leishmania) infantum e transmitida por flebotomíneos e é uma das principais zoonoses do Brasil que se encontra em expansão em estados da região sul do país, sendo os cães o principal reservatório urbano do parasito. O presente estudo investigou a ocorrência de LVC em dois municípios, Erval Velho e Herval d’Oeste localizados no meio-oeste de Santa Catarina. Para tanto, amostras de sangue periférico de 126 cães foram coletadas em ambas as cidades e submetidas à detecção de anticorpos anti-L. infantum por meio de testes de ELISA e imunofluorescência indireta (IFI), bem com a detecção de k-DNA pela reação em cadeia de polimerase (PCR). Além disso, também foram observados os sintomas clínicos e as condições ambientais associadas a esses animais. Dos cães examinados, 35,7% (45/126) foram positivos para pelo menos um dos três testes, dois cães (1,6%) foram positivos em todos os três testes, 12 cães (9,5%) foram positivos tanto no ELISA quanto na IFI, enquanto 21 cães (16,7%) foram positivos para ELISA e 15 (11,9%) para o IFI. A amplificação do k-DNA de L. infantum foi positiva em 9 dos 126 cães (7,1%). Entre os cães positivos seis apresentaram um ou mais sintomas clínicos correlacionados com a LVC. Esses resultados confirmaram a ocorrência de LVC na região e destacaram a importância do monitoramento e implementação de medidas de controle para a LVC nessa região

Abstract Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum is transmitted by phlebotomine sandflies and a major zoonotic disease in Brazil. Due to the southward expansion of the disease within the country and the central role of dogs as urban reservoirs of the parasite, we have investigated the occurrence of CVL in two municipalities Erval Velho and Herval d’Oeste in the Midwest region of Santa Catarina state. Peripheral blood samples from 126 dogs were collected in both cities and tested for anti-L. infantum antibodies by indirect enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence reaction (IIF) and for the presence of parasite DNA by polymerase chain reaction (PCR) in peripheral blood. From examined dogs, 35.71% (45/126) were positive for at least one of the three tests and two (1.6%) were positive in all performed tests. Twelve dogs (9.5%) were positive for both ELISA and IIF, while 21 dogs were exclusively positive for ELISA (16.7%), and 15 (11.9%) for IIF. L. infantum k-DNA was detected by PCR in 9 out of 126 dogs (7.1%) and clinical symptoms compatible with CVL were observed for 6 dogs. Taken together, these results indicate the transmission of CVL in this region, highlighting the needs for epidemiological surveillance and implementation of control measures for CVL transmission in this region.

RESUMO Introdução: A proteína CRISP3 é expressa em baixos níveis na próstata humana normal, mas superexpressa no câncer de próstata (CaP). Contudo, sua relevância em pacientes com CaP ainda não está clara. O teste de antígeno específico da próstata (PSA) pode proporcionar interpretações erradas em determinadas circunstâncias, resultando em excesso de tratamento para tumores indolentes. Novos biomarcadores são fundamentais para evitar tratamentos desnecessários e melhorar a qualidade de vida do paciente. Objetivo: Avaliar se a expressão de CRISP3 poderia ser um bom biomarcador para CaP. Métodos: A expressão de CRISP3 foi determinada por imuno-histoquímica em seções de tecido de CaP de 25 pacientes submetidos a prostatectomia radical. O sistema de classificação Gleason foi utilizado como indicador prognóstico, e o estadiamento foi determinado pelo sistema TNM. Parâmetros clínicos e níveis de PSA antes e pós-cirurgia foram determinados. Resultados: A expressão de CRISP3 foi forte em 14 (56%) amostras; moderada em quatro (16%) e fraca em sete (28%). Não houve correlação entre a expressão de CRISP3 e PSA pré e pós-tratamento. Quinze (60%) biópsias de CaP apresentaram extensão do tumor primário pT2. Sete pacientes (28%) mostraram escore de Gleason maior que 7; treze (52%), igual a 7; e cinco (20%), menor que 7. Não houve diferenças estatísticas significativas entre o escore de Gleason e a expressão de CRISP3. Conclusão: CRISP3 é expresso no CaP em diferentes níveis. Estudos adicionais são necessários para avaliar se CRISP3 realmente pode ser usado como biomarcador.

ABSTRACT Introduction: Cysteine-rich secretory protein 3 (CRISP3) is expressed at low levels in normal human prostate but often overexpressed in prostate cancer (PCa). The relevance of this overexpression for the malignancy of PCa is still unclear. The prognostic value of the currently used prostate specific antigen (PSA) test can be misleading under certain circumstances, resulting in overtreatment of indolent tumors. New biomarkers are needed to reduce overtreatment and improve quality of life of men. Objective: Evaluate if CRISP3 expression could be a good biomarker for PCa. Methods: CRISP3 expression was determined by immunohistochemistry in tissue sections of prostate cancer from twenty-five patients subjected to radical prostatectomy. Gleason grading system was used as prognostic indicator and the staging of PCa was defined using the TNM system. Clinical parameters and PSA levels before and after surgery were determined. Results: CRISP3 expression was strong in 14 (56%), moderate in four (16%) and weak in seven (28%) specimens. There was no correlation between the intensity of CRISP3 expression and pre- and post-treatment PSA levels. Fifteen (60%) of PCa biopsies showed extension of the primary tumor pT2. Seven patients (28%) showed Gleason score higher than 7; thirteen (52%) equal to 7, and five (20%) lower than 7. There were no significant statistical differences between Gleason score and CRISP3 expression. Conclusion: CRISP3 is expressed in prostate cancer at different levels. Additional studies are required to better evaluate if CRISP3 could be used as a biomarker.

RESUMEN Introducción: La proteína rica en cisteína secretora 3 (CRISP3) se expresa en bajos niveles en la próstata humana normal, pero está mayormente expresada en el cáncer de próstata (CaP). Todavía, su relevancia en pacientes con CaP aún no está clara. La prueba de antígeno prostático específico (PSA) puede generar interpretaciones erróneas bajo ciertas circumstancias, acarreando sobretratamiento de tumores indolentes. Nuevos biomarcadores son fundamentales para evitar tratamientos innecesarios y mejorar la calidad de vida del paciente. Objetivo: Evaluar se la expresión de CRISP3 podría ser un buen biomarcador de CaP. Métodos: Se determinó la expresión de CRISP3 por inmunohistoquímica en cortes de tejido de CaP de 25 pacientes sometidos a prostatectomía radical. La clasificación Gleason fue utilizada como indicador pronóstico, y la estadificación fue determinada por el sistema TNM. Parámetros clínicos y niveles de PSA antes y después de la cirurgía fueron determinados. Resultados: La expresión de CRISP3 fue fuerte en 14 (56%) muestras; moderada en cuatro (16%) y débil en siete (28%). No hubo relación entre la expresión de CRISP3 y PSA pre y post-tratamiento. Quince (60%) biopsias de CaP tuvieron extensión del tumor primario pT2. Siete pacientes (28%) demostraron escala de Gleason mayor que 7; trece (52%), igual a 7; y cinco (20%), menor que 7. No hubo diferencias estadísticas significativas entre la escala de Gleason y la expresión de CRISP3. Conclusión: CRISP3 se expresa en CaP en diferentes niveles. Se necesitan estudios adicionales para evaluar se CRISP3 puede realmente ser usado como biomarcador.

ABSTRACT This paper presents a paleoenvironmental reconstruction from palynological analyses of a sedimentary core of Holocene age, drilled at municipality of Garopaba (Santa Catarina), Southern Brazil. A total of 46 samples was collected for palynological analyses in the 450 cm-long core PCSC-3, as also three samples for radiocarbon dating and granulometric analyses. The palynological content includes 84 taxa related to pollen grains of angiosperms (38) and gimnosperm (3), spores of pteridophyta (16) and bryophyta (2), spores of fungi (8), algae (3), acritarchs (3), dinoflagellate cysts (2) and microforaminiferal linings (1). Three specimens of acritarchs are described and illustrated in detail. Three palynological phases were defined based on changes in assemblages: Phase I, Phase II and Phase III. The Phase I is characterized as a lagoonal paleoenvironment with marine influence from the beginning of the sedimentation (5390 cal yr BP), based on occurrences of acritarchs, dinoflagellate cysts and microforaminiferal linings. The Phase II (3032 yr BP until 858 cal yr BP) also is characterized by a lagoonal paleoenvironment, however, presented decrease in percentage of marine elements and increase in freshwater algae record, suggesting less marine influence in the lagoonal body. In Phase III (last 856 years), underwater sedimentation prevailed, under swamp-like conditions.

Resumo O período crepuscular é um dos fatores que pode influenciar na atividade hematofágica dos mosquitos. Muitos desses insetos iniciam ou terminam suas atividades nesse período. O objetivo deste trabalho foi estudar os Culicidae que ocorrem no crepúsculo vespertino em uma área de Floresta Atlântica no oeste de Santa Catarina, sul do Brasil. Além disso, foi analisada a possível influência de fatores abióticos, bem como abundância e riqueza de espécies. Para melhor avaliar a influência do período crepuscular na composição das espécies e na abundância destas, o crepúsculo foi dividido em três períodos: pré-crepúsculo, crepúsculo e pós-crepúsculo. Ao final do estudo foram coletados 984 exemplares distribuídos em 12 gêneros e 23 espécies. Trichoprosopon pallidiventer (Lutz, 1905) (59,76%), Aedes crinifer (Theobald, 1903) (8,13%) e Ae. scapularis (Rondani, 1848) (5,89%) foram as espécies mais abundantes. A maior abundância e riqueza de espécies se deram na primavera. Dentre os três períodos estudados, o pré-crepúsculo apresentou a maior abundância de mosquitos, em contrapartida, o pós-crepúsculo apresentou a menor abundância. Os períodos pré-crepuscular e crepuscular apresentaram maior similaridade entre si com relação à composição das espécies. Relacionando os fatores abióticos e as três espécies mais abundantes, não foi observada correlação significativa nos dados avaliados nos sete e 15 dias anteriores às coletas. Entretanto, a temperatura apresentou uma correlação positiva para estas três espécies. A relação entre as espécies coletadas e a potencial transmissão de agentes etiológicos causadores de doenças foi comentada.

Abstract Crepuscular period is one of the factors that may influence the biting activity of mosquitoes. Many of these insects have a peak activity in this period. The purpose of this study was to investigate the afternoon crepuscular activity of Culicidae in a remaining area of Atlantic Forest in western Santa Catarina, southern Brazil. Moreover, the possible influence of abiotic factors, the abundance and species richness were verified. In order to better analyze the influence of crepuscular period in specific composition and abundance of mosquitoes, the dusk was divided into three periods: pre-sunset, sunset and post-sunset. At the end of the study, nine hundred and eight four specimens distributed in 12 genera and 23 species were collected. Trichoprosopon pallidiventer (Lutz, 1905) (59.76%), Aedes crinifer (Theobald, 1903) (8.13%), Ae. scapularis (Rondani, 1848) (5.89%) were the most abundant species. Spring time presented the greatest abundance and species richness. During the study, among the three periods evaluated, pre-sunset had the greatest abundance and post-sunset the lowest. Pre-sunset and sunset had the greatest similarity between species. Regarding to the abiotic factors evaluated seven and 15 days before sampling, they did not present significant correlation for the three most abundant species. However, temperature had a positive correlation to these species. Moreover, the correlation between collected species and its possible role as vectors of etiological agents of diseases was discussed.

We report herein a study on the glycosylation of cyclohexanol with four D-glucosamine-based peracetylated glycosyl chlorides bearing different substituents at C-2 and three glycosylation promoters, silver carbonate, silver triflate and mercury II chloride/mercury II oxide, by the Koenigs-Knorr method. Under the conditions studied, glycosylation was successful only when 3,4,6-tri-O -acetyl-2-deoxy-2-phthalimido-α-D-glucopyranosyl chloride was used as the glycosyl donor, with silver carbonate proving the best promoter. In order to investigate the influence of the nature of the halogen at C-1, we also carried out the glycosylation of cyclohexanol with 3,4,6-tri-O -acetyl-2-deoxy-2-phthalimido-α-D-glucopyranosyl bromide, a more reactive glycosyl donor. As expected, the yield with the bromide derivative was higher with the three promoters and, again, silver carbonate was the most efficient promoter. Finally, to illustrate the well-known efficient procedure for conversion of the phtalimido group at C-2 to the corresponding acetamido group, cyclohexyl 3,4,6-tri-O -acetyl-2-deoxy-2-phtalimido-β-D-glucopyranoside was converted into cyclohexyl 2-deoxy-2-acetamido-β-D-glucopyranoside in two steps, namely, hydrazinolysis of the phtalimido group followed by chemoselective acetylation of the free amino group by treatment with acetic anhydride in methanol, at 77% overall yield.

Este trabalho apresenta a integração da análise micropa leontológica (palinologia e foraminíferos) e de isótopos (razão 87Sr/86Sr) de um intervalo selecionado do poço 2-TG-96-RS, perfurado na porção onshore da Bacia de Pelotas, Rio Grande do Sul, Brasil. Um total de oito amostras da seção entre 140,20 e 73,50 m de profundidade foi selecionado para análise palinológica, revelando associações de palinomorfos abundantes e diversificados. Espécies de esporos, grãos de pólen, cistos de dinoflagelados são os palinomorfos mais comuns encontrados. Foraminíferos calcáreos bentônicos e planctônicos foram recuperados dos dois níveis mais inferiores da seção (140,20 e 134,30 m). Com base na amplitude estratigráfica de espécies de cistos de dinoflagelados e esporomorfos, um intervalo de idade entre o Mioceno Superior e o Plioceno Inferior é atribuído. A idade relativa obtida a partir da razão 87Sr/86Sr em carapaças de foraminíferos calcários indica correspondência no Mioceno Final (Messiniano), corroborando o posicionamento bioestratigráfico realizado com palinomorfos. Interpretações paleoambientais basea das na distribuição quantitativa dos componentes orgânicos (palinomorfos, fitoclastos e matéria orgânica amorfa) ao longo da seção e na associação de foraminíferos indicam ambiente marinho raso para a seção. Dois intervalos palinológicos foram reconhecidos com base na análise de palinofácies, relacionados a condições de plataforma média a externa (140,20 a 128,90 m) e de plataforma interna (115,75 a 73,50 m).

This paper presents the integration of micropaleontological (palynology and foraminifera) and isotopic (87Sr/86Sr) analysis of a selected interval from the well 2-TG-96-RS, drilled on the onshore portion of the Pelotas Basin, Rio Grande do Sul, Brazil. A total of eight samples of the section between 140.20 and 73.50 m in depth was selected for palynological analysis, revealing diversified and abundant palynomorph associations. Species of spores, pollen grains and dinoflagellate cysts are the most common palynomorphs found. Planktic and benthic calcareous foraminifera were recovered from the lowest two levels of the section (140.20 and 134.30 m). Based on the stratigraphic range of the species of dinoflagellate cysts and sporomorphs, a span age from Late Miocene to Early Pliocene is assigned. The relative age obtained from the 87Sr/86Sr ratio in shells of calcareous foraminifers indicates a Late Miocene (Messinian) correspondence, corroborating the biostratigraphic positioning performed with palynomorphs. Paleoenvironmental interpretations based on the quantitative distribution of organic components (palynomorphs, phytoclasts and amorphous organic matter) throughout the section and on foraminiferal associations indicate a shallow marine depositional environment for the section. Two palynologicals intervals were recognized based on palynofacies analysis, related to middle to outer shelf (140.20 to 128.90 m) and inner shelf (115.75 to 73.50 m) conditions.

Pneumonia due to herpes simplex virus type 1: A difficult to diagnose condition with a poor prognosis in lung transplants Abstract Herpetic pneumonia is a rare disease that mostly affects people with immune deficiency whose outcome is frequently fatal. Its appearance is described in the first two months after surgery. In our experience it was evidenced in the post transplant apart since 10 months, 1 ½ years, 8 years and 19 years after the transplant. We described the CT findings detected in 4 lung transplant recipient that had evolved with herpetic pneumonia. Patients presented fever, cough, sputum and dyspnea with progressive worsening of functional class, four died after the disease. The histological diagnosis is made by bronchoalveolar lavage or transbronchial biopsy with subsequent staining with hematoxylin - eosin and / or immunohistochemistry procedure. The highlighted CT findings were ground-glass opacities patchy distribution, airspace consolidation, pleural effusion and bronchiectasis. Due to its low frequency and poor prognosis is important to know this entity in lung transplant recipients to avoid a delayed diagnosis and treatment.

La neumonía herpética es una rara enfermedad que afecta casi con exclusividad a personas con un déficit de inmunidad. Con frecuencia tiene un desenlace fatal. Su aparición está descrita en los primeros 2 meses luego del trasplante pulmonar, pero en nuestra experiencia se evidenció después de los 10 meses, 1 año y medio, 8 y 19 años. Describimos los hallazgos tomográficos detectados en 4 pacientes con trasplantes pulmonares que cursaron neumonía herpética. Presentaban fiebre, tos, expectoración y disnea, con empeoramiento progresivo de su clase funcional, y todos fallecieron tras la aparición de la enfermedad. El diagnóstico histológico se realiza mediante lavado bronquioalveolar o biopsia transbronquial, con posterior tinción con hematoxilina-eosina y/o marcación inmunohistoquímica. Los hallazgos tomográficos destacados fueron: opacidades en vidrio esmerilado de distribución parcheada, consolidación del espacio aéreo, derrame pleural y bronquiectasias. Debido a su baja frecuencia y mal pronóstico, es importante conocer y tener presente esta entidad en personas trasplantadas de pulmón para no demorar el diagnóstico y actuar lo más rápidamente posible.

Neste estudo retrospectivo (1980-1998), analisamos clínico-demograficamente, a partir dos prontuários do Hospital Universitário de Fortaleza (Brasil), um grupo de 87 pacientes que apresentavam sinais e sintomas de doenças do neurônio motor (DNMs). Eles foram diagnosticados clinicamente, e através de exames complementares. Para o diagnóstico da esclerose lateral amiotrófica (ELA), usamos os critérios da Federação Mundial de Neurologia. A análise clínico-demográfica dos 87 casos de DNMs evidenciou a existência de 4 casos de atrofia muscular espinhal (AME), 5 casos de variantes da ELA: 2 de paralisia bulbar progressiva (PBP), 2 de atrofia muscular progressiva (AMP) e 1 de amiotrofia monomélica (AM), e 78 casos de ELA. Esses últimos eram constituídos de 51 homens e 27 mulheres, com uma idade média de 42,02 anos. Eles foram subdivididos em 4 grupos etários: de 15 a 29 anos (n= 17), de 30 a 39 anos (n= 18), de 40 a 69 anos (n= 39) e de 70 a 78 anos (n= 4). Dos 78 casos de ELA, 76 eram da forma esporádica clássica, enquanto que apenas 2 da forma familiar. A análise dos 87 pacientes com DNMs do Hospital Universitário de Fortaleza mostrou predomínio de pacientes com ELA, com um número elevado de casos de ELA juvenil e da forma adulta de início precoce.

In this retrospective (1980-1998) study, we have analyzed clinico-demographically, from the records of the University Hospital of Fortaleza (Brazil), a group of 87 patients showing signs and symptoms of motor neuron diseases (MNDs). Their diagnosis was determined clinically and laboratorially. The WFN criteria were used for amyotrophic lateral sclerosis (ALS) diagnosis. The clinico-demographic analysis of the 87 cases of MNDs showed that 4 were diagnosed as spinal muscular atrophy (SMA), 5 cases as ALS subsets: 2 as progressive bulbar paralysis (PBP), 2 as progressive muscular atrophy (PMA) and 1 as monomelic amyotrophy (MA), and 78 cases of ALS. The latter comprised 51 males and 27 females, with a mean age of 42.02 years. They were sub-divided into 4 groups according to age: from 15 to 29 years (n= 17), 30 to 39 years (n= 18), 40 to 69 years (n= 39) and 70 to 78 years (n= 4). From the 78 ALS patients, 76 were of the classic sporadic form whilst only 2 were of the familial form. The analysis of the 87 patients with MNDs from the University Hospital of Fortaleza showed a predominance of ALS patients, with a high number of cases of juvenile and early onset adult sporadic ALS.

Relatamos os aspectos clínicos e eletrofisiológicos preliminares de um paciente hospitalizado que possivelmente apresenta crises de epilepsia parcial contínua (Kojevnikov). Discutimos as diferentes etiologias dessa condição e salientamos a possível forma idiopática, nesse caso.

We report on the preliminary clinical and electrophysiological aspects of an in-patient possibly presenting epilepsia partialis continua (Koshevnikov). We discuss the different etiologies and emphasize on the possible idiopathic form in this case.

Tools for the genetic manipulation of Trypanosoma cruzi are largely unavailable, although several vectors for transfection of epimastigotes and expression of foreign or recombinant genes have been developed. We have previously constructed several plasmid vectors in which recombinant genes are expressed in T. cruzi using the rRNA promoter. In this report, we demonstrate that one of these vectors can simultaneously mediate expression of neomycin phosphotransferase and green fluorescent protein when used to stably transfect cultured epimastigotes. These stably transfected epimastigotes can be selected and cloned as unique colonies on solid medium. We describe a simple colony PCR approach to the screening of these T. cruzi colonies for relevant genes. Thus, the methodologies outlined herein provide important new tools for the genetic dissection of this important parasite.

The diagnosis of tick-borne diseases such as babesiosis still depends on observing the parasite in the infected erythrocyte. Microscopic observation is tedious and often problematic in both early and carrier infections. Better diagnostic methods are needed to prevent clinical disease, especially when susceptible cattle are being moved into disease enzootic areas. This study evaluates two techniques for early diagnosis of Babesia bovis infections in cattle, DNA probes specific for the organism and fluorescent probes specific nucleic acid. The radioisotopically labeled DNA probes are used in slot blot hybridizations whith lysed blood samples, not purified DNA. Thusfar, the probe is specific for B. bovis and can detect as few as 1000 B. bovis parasites in 10µl of blood. The specificity of the fluorescent probe depends on the characteristic morphology of the babesia in whole blood samples, as determined microscopically. The fluorescent probe detects as afew as 10,000 B. bovis parasites in 10*l as blood. The application of each method for alboratory and field use is discussed.