Quantification of Listeria spp. contamination on shell and flesh of cooked black tiger prawns (Penaeus monodon)

Aims: To quantify Listeria levels on the shell and flesh of artificially contaminated cooked prawns after peeling, and determine the efficacy of Listeria innocua as a model for L. monocytogenes in this system. Methods and Results: A L. monocytogenes and L. innocua strain were inoculated separately onto cooked black tiger prawns using two protocols ( immersion or swabbing with incubation). Prawns were peeled by two methods ( gloved hand or scalpel and forceps) and numbers of Listeria on shells, flesh and whole prawn controls were determined. Prawns were exposed to crystal violet dye to assess the penetration of liquids. Regardless of preparation method or bacterial strain there were ca 1log(10) CFU more Listeria per shell than per peeled prawn. Dye was able to penetrate to the flesh in all cases. Conclusions: Shell-on prawns may be only slightly safer than shell-off prawns. Listeria innocua is an acceptable model for L. monocytogenes in this system. Significance and Impact of the Study: Reduced risk from L. monocytogenes on prawns can only be assured by adequate hygiene or heating.

Biologia de Anopheles (Kerteszia) neivai H., D. & K., 1913 (Diptera: Culicidae) en la costa Pacífica de Colombia: I. Fluctuación de la población larval y características de sus criaderos

En Charambirá, localidad del município de Istmina (Chocó) en la costa Pacífica de Colombia, donde existe un problema de malaria endémica, se estudió la fluctuación de la población larval y las características de los criaderos de Anopheles (Kerteszia) neivai, especie considerada como posible vectora de malaria en esa región del país. La vegetación circundante fue dividida en tres estratos de acuerdo al grado de cobertura foliar. Se demarcaron cuadrantes de 100 m² en cada estrato para el muestreo de plantas epífitas de la família Bromeliaceae, en las cuales se acumula agua que sirve como criadero para esta especie de anofelino. Se tomaron datos de temperatura, pH y volumen del agua contenida en cada bromelia. El mayor número de larvas se detectó en el estrato 1 (manglar) a una altitud inferior a 4 m, pero no se encontró evidencia significativa de estratificación vertical de la población larval de A. neivai hasta los 8 m. Se evidenció una correlación lineal positiva entre el número de larvas y el volumen de agua contenida en cada bromelia; por otra parte se observó también una correlación directa entre la precipitación mensual acumulada y la fluctuación poblacional de esta especie. Los índices larvales mas altos se observaron entre los meses de marzo a abril y de julio a agosto. La mortalidad larval fue alta en el primer estadío (43,5%) y solo un 23,7% sobrevivió hasta el cuarto.

Avaliação do desenvolvimento larval de Anopheles darlingi criado em laboratório sob diferentes dietas

São testadas três dietas para larvas de A. darlingi, buscando-se os seguintes parâmetros indicativos de desenvolvimento nesta fase: tempo de evolução total e de cada estádio, sobrevivência ao estádio, diária e total. A metodologia utilizada na determinação desses parâmetros constitui-se da combinação de dois métodos de análise da estatística vital, adaptados ao estudo de populações mantidas em laboratório. Os tempos de evolução total e de cada estádio foram determinados gráficamente a partir das curvas de tendência dos estádios medianos da colônia, em inspeções sucessivas. Os valores de sobrevivência diária, ao estádio e total foram calculados a partir de tábuas de sobrevivência. Os resultados permitiram eleger a dieta composta de uma parte de farinha de peixe para duas partes de farinha de pão e duas partes de germe de trigo como a mais adequada ao desenvolvimento larval, com duração de 12,9 dias entre o primerio estádio e a emersão do adulto e sobrevivência total de 95%.

Duration of larval and pupal development stages of Aedes albopictus in natural and artificial containers

Aedes albopictus were reared in different containers: a tree hole, a bamboo stump and an auto tire. The total times from egg hatching to adult emergence were of 19.6,27.3 and 37.5 days, respectively, according to the container. The first, second and third-instar larvae presented growth periods with highly similar durations. The fourth-instar larvae was longer than the others stages. The pupation time was longer than the fourth-instar larvae growth period. The temperature of the breeding sites studied, which was of 18° C to 22° C on average, was also taken into consideration. The mortality of the immature stages was analysed and compared as between the experimental groups; it was lower in the natural containers than in the discarded tire. The average wing length of adult females emerging from tree hole was significantly larger (p < 0.05) than that of those emerging from the tire.

Human case of gastric infection by a fourth larval stage of Pseudoterranova decipiens (Nematoda, Anisakidae)

Only three cases of human infection by anisakid nematodes have been reported in Chile since 1976. In the present case, an anisakid worm, identified as a fourth-stage Pseudoterranova decipiens larva, was removed with a gastroendoscopic biopsy clipper from the stomach of a 45 year-old man from southern Chile. The patient, who presented acute epigastric pain and a continuous sensation of having an empty stomach, reported having eaten smoked fish. The worm was fixed in 70% ethanol and cleaned in lactophenol for morphological study. The morphometric characteristics of the worm are described and drawn. Anisakid larvae in fish flesh can be killed by freezing or cooking.

Susceptibilidade larval de duas populações de Aedes egypti a inseticidas químicos

OBJETIVO: A susceptibilidade dos insetos tem sido um dos mais importantes aspectos a ser monitorados em programas de saúde pública que tratam do controle de vetores. O estudo objetiva avaliar a susceptibilidade de larvas de Aedes aegypti a inseticidas químicos em áreas sujeitas ou não a controle. MÉTODOS: Bioensaios foram realizados com concentração de diagnóstico e concentração múltipla, segundo padrão da Organização Mundial de Saúde para as coletas de larvas de Aedes aegypti, em uma área não sujeita -- Campinas, SP -- e em uma outra área sujeita -- Campo Grande, MS -- a tratamentos químicos de controle. RESULTADOS: Larvas de Aedes aegypti coletadas em Campinas indicaram resistência potencial à concentração-diagnóstico (CD) de 0,04 ppm do organofosforado temephos. O teste de concentração múltipla registrou sobrevivência de 24,5% à concentração de 0,0125 ppm. A susceptibilidade dessa mesma linhagem foi avaliada para o organofosforado fenitrothion (CD=0,08 ppm) e o piretróide cipermetrina (CD=0,01 ppm), resultando em valores normais para essas concentrações. Larvas de Ae. aegypti coletadas em Campo Grande mostraram susceptibilidade normal ao temephos (CD=0,04 ppm) e à cipermetrina (CD=0,01 ppm). Também foram estabelecidas as CL50 e as CL95 de cipermetrina 25 CE, cyfluthrin 5 CE, betacyfluthrin 1,25 SC e propoxur 20 CE para Ae. aegypti. Com base nos dados da linhagem-padrão Rockefeller, foram estimadas as razões de resistência de 2,9, 2,2, 2,4 e 1,3, respectivamente, pela CL50, e de 3,5, 2,6, 3,9 e 1,3 pela CL95. CONCLUSÃO: Os resultados reforçam a necessidade de avaliações prévias e monitoramento da efetividade dos inseticidas que devem ser usados em programas de controle de mosquitos.

Susceptibilidade larval de populações de Aedes aegypti e Culex quinquefasciatus a inseticidas químicos

OBJETIVO: Avaliar a susceptibilidade a inseticidas químicos de larvas de Culex quinquefasciatus e Aedes aegypti, provenientes de áreas sujeitas ou não a tratamentos de controle. MÉTODOS: Foram coletadas larvas de mosquitos em uma área não sujeita a tratamentos com inseticidas (Campinas, SP) e em áreas sujeitas a esses tratamentos (Campo Grande, MS e Cuiabá, MT). Foram usados bioensaios com concentrações diagnóstico e concentrações múltipla de inseticidas organofosforados e piretróides, segundo padrão da Organização Mundial de Saúde, para avaliar a susceptibilidade dessas larvas. RESULTADOS: Ensaios com larvas de Culex quinquefasciatus de Campinas, SP, permitiram a suspeita de resistência à cipermetrina e evidenciaram resistência à ciflutrina. Larvas dessa espécie coletadas em Campo Grande, MS, e Campinas, SP, apresentaram resistência ao temephos. Para a colônia campineira desta espécie, foram estabelecidas as razões de resistência: RR50=6,36 e RR95=4,94, com base em linhagem susceptível padrão. Adicionalmente, os testes com Aedes aegypti mostraram susceptibilidade similar ao temephos em uma população de campo (Cuiabá, MT) e uma de laboratório. CONCLUSÕES: Os resultados indicam resistência a organofosforado e piretróides em Culex quinquefasciatus, evidenciando a necessidade de avaliações e monitoramento da efetividade dos inseticidas a serem usados nos programas de controle de mosquitos.

Sensibility of different larval stages of Schistosoma mansoni to the Larvae Disappearing Reaction (LDR) in murine schistosomiasis

The delay produced by drug, in the process of cercaria-schistosomulum transformation, was used to verify the sensibility of different larval stages to the host cell immune responses, in vivo. The peritoneal cavity of mice, a model used for in vivo observations, was choiced for the experiments. As well characterized schistosomules, cercariae and larvae in the process of transformation were coated and arrested by host cells, and could not be recovered by simple saline washings. After 10-²M EDTA saline washings, they were released alive, with good vitality and movements. Thus, different kind of larvae in the process of adaptation of the cercaria to the host are strongly coated by immune cells, but these fail to kill the invading organisms, at least during a few hours after invasion.

An unusual ground larval habitat of Aedes albopictus

Relata-se o encontro de criadouro de Aedes albopictus no solo. Tratou-se de buraco resultante da queda de árvore Piptadenia ("angico branco") onde foram coletadas larvas daquele mosquito, juntamente com representantes de Culex declarator, Cx. quinquefasciatus e Culex (Cux.) sp. do Grupo Coronator. O criadouro comportou cerca de 17 litros de água proveniente de chuvas locais. O achado deu-se em mata residual dentro de perímetro urbano da cidade de Pindamo-nhangaba no Vale do Paraíba, Estado de São Paulo, Brasil. Discute-se o significado do encontro como provável indicador de ecletismo comportamental e de adaptação do culicídeo ao ambiente antrópico.

Schistosoma mansoni and other larval trematodes in Biomphalaria tenagophila (Planorbidae) from Guarulhos, São Paulo State, Brazil

A total of 909 Biomphalaria tenagophila were collected from two areas in Guarulhos (Metropolitan area of São Paulo, São Paulo State, Brazil) to assess larval trematode infections. In all collection sites, only this species was found and 183 (20.13%) harbored trematode infections. In these collections, four morphologically distinguishable types of cercariae were identified by confocal microscopy. Xiphidiocercaria (Cercaria lutzi) was the most common type of cercaria recovered, contributing 76.5% of all infections. Schistosoma mansoni cercariae were recovered and comprised the total of 13.11%. Strigea cercaria (Cercaria caratinguensis) and Brevifurcate pharyngeate Clinostomatoide cercaria (Cercaria ocellifera) contributed 8.33% and 2.22% of all infections, respectively. Double infections (S. mansoni and C. lutzi) were found in twelve snails, contributing 6.55% of all infections. In all sites studied, small vertebrates were found in snail habitats and it was observed human contact with the water. The presence of trematode infected snails in large cities has public health implications. It further provides a starting point for some comprehensive studies on snail-related aspects of transmission and biology of trematode of medical and veterinary importance.

EFFICACY OF NITAZOXANIDE AGAINST Toxocara canis: LARVAL RECOVERY AND HUMORAL IMMUNE RESPONSE IN EXPERIMENTALLY INFECTED MICE

SUMMARY The efficacy of nitazoxanide (NTZ) against toxocariasis was investigated in an experimental murine model and results were compared to those obtained using mebendazole. Sixty male BALB/c mice, aged six to eight weeks-old, were divided into groups of 10 each; fifty were orally infected with 300 larvaed eggs of T. canisand grouped as follows, G I: infected untreated mice; G II: infected mice treated with MBZ (15 mg/kg/day) 10 days postinfection (dpi); G III: infected mice treated with NTZ (20 mg/kg/day) 10 dpi; G IV: infected mice treated with MBZ 60 dpi; G V: infected mice treated with NTZ 60 dpi; GVI: control group comprising uninfected mice. Mice were bled via retro-orbital plexus on four occasions between 30 and 120 dpi. Sera were processed using the ELISA technique to detect IgG anti- Toxocaraantibodies. At 120 dpi, mice were sacrificed for larval recovery in the CNS, liver, lungs, kidneys, eyes and carcass. Results showed similar levels of anti- ToxocaraIgG antibodies among mice infected but not submitted to treatment and groups treated with MBZ or NTZ, 10 and 60 dpi. Larval recovery showed similar values in groups treated with NTZ and MBZ 10 dpi. MBZ showed better efficacy 60 dpi, with a 72.6% reduction in the parasite load compared with NTZ, which showed only 46.5% reduction. We conclude that administration of these anthelmintics did not modify the humoral response in experimental infection by T. canis. No parasitological cure was observed with either drug; however, a greater reduction in parasite load was achieved following treatment with MBZ.

ANTHELMINTIC ACTIVITY OF CHLOROPHYLLIN AGAINST DIFFERENT LARVAL STAGES OF Fasciola gigantica

Fasciolosis is a food borne zoonosis, caused by the digenetic trematode Fasciola. Freshwater lymnaeid snails are the intermediate host of the trematodes. Chlorophyllin, a semi-synthetic derivative of chlorophyll and its formulations obtained from freeze dried cow urine (FCU) had their toxicity tested against redia and cercaria larvae of F. gigantica. The larvicidal activity of chlorophyllin and its formulations were found to depend on both, time and concentration used against the larvae. Toxicity of chlorophyllin + FCU (1:1 ratio) in sunlight against redia larva (8 h LC50: 0.03 mg/mL) was more pronounced than using just chlorophyllin (8 h LC50: 0.06 mg/mL). Toxicity of chlorophyllin + FCU in sunlight against redia (8 h LC50: 0.03 mg/mL) was higher than against cercaria (8 h LC50: 0.06 mg/mL). The larvicidal activity of chlorophyllin in sunlight (redia/cercaria larvae: 8 h LC50: 0.06 mg/mL) was more pronounced than under laboratory conditions (redia: 8 h LC50: 22.21 mg/mL/, cercaria 8 h LC50: 96.21 mg/mL). Toxicity of FCU against both larvae was lower than that of chlorophyllin and chlorophyllin + FCU. Chlorophyllin and its formulations + FCU were 357.4 to 1603.5 times more effective against redia/cercaria larvae in sunlight than under laboratory conditions. The present study has shown that chlorophyllin formulations may be used as potent larvicides against fasciolosis.

Chemotherapeutic effects on larval stages of Schistosoma mansoni during infection and re-infection of mice

The sensitivity of the larval stages of Schistosoma mansoni to chemotherapy with praziquantel and oxamniquine was tested in mice during primary and secondary infections and after different intervals from cercarial exposure. Worm recovery by perfusion of the porto-mesenteric system, followed by counting and a morphometric study of the parasite, allowed the conclusion that the relative resistance of the larval stages of S. mansoni to schistosomicide drugs, demonstrated in primary infections, also persists when the host is already infected. This indicates that a therapeutic failure may result when an infected host is treated some time after being re-infected, because of the presence of migrating, drug-resistant, immature forms of the parasite.

Efeito da densidade larval no tamanho de adultos de Aedes aegypti criados em condições de laboratório

Para avaliar a influência da densidade larval no desenvolvimento do mosquito Aedes aegypti, foram testadas três diferentes densidades. Observou-se que em condições de alta densidade larvária e carência de alimentos houve uma redução no tamanho dos adultos e aumento na taxa de mortalidade de Aedes aegypti em condições de laboratório.