Larvicidal & emergence inhibitory activities of NeemAzal T/S 1.2 per cent EC against vectors of malaria, filariasis & dengue.

Background & objectives: Vector control, using agents of chemical origin, continues to be practiced in the control of vector borne diseases. However, due to some drawbacks including lack of selectivity, environmental contamination, and emergence and spread of vector resistance, development of natural products for vector control has been a priority in this area. In the present study we evaluated the larvicidal and emergence inhibitory activities of a neem based formulation Neem Azal T/S 1.2 per cent EC against the vectors of malaria, filariasis and dengue. Method: Larvicidal and emergence inhibition (EI) activity of a neem formulation, NeemAzal T/S 1.2 per cent EC, was studied in the laboratory respectively against early 4th and early 3rd instar larvae of Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti following standard procedures. Results: Among the three vector species studied, An. stephensi was highly susceptible to NeemAzal T/S as revealed by the LC50 and LC90 values (1.92 and 2.76 ppm). The formulation produced an overall mortality or inhibition of emergence of 90 per cent (EI90, when 3rd instar larvae were treated) at 0.046, 0.208 and 0.866 ppm in An. stephensi, Cx. quinquefasciatus and Ae. aegypti, respectively. The corresponding EI50 values were 0.006, 0.048 and 0.249 ppm. On treatment, NeemAzal T/S induced certain morphogenetic abnormalities, broadly characterized in five types, in larvae, pupae and adults of all the three vector species. The percentage of dead specimens of any stage showing morphogenetic abnormalities was the maximum in Cx. quinquefasciatus (14.4%; n=2113) followed by Ae. aegypti. Interpretation & conclusions: Our results indicated that because of its emergence inhibition activity, NeemAzal T/S 1.2 per cent EC could be a promising candidate for the use in integrated vector management programme and replace chemical insecticides.

Intravitreal live adult Brugian filariasis.

Human ocular infestation by live filarial worm is a rare occurrence and has been reported mostly form South-East Asia. It involves the eyelids, conjunctiva, cornea, anterior chamber and uvea. No case of intravitreal Brugian microfilaria / adult worm has been found by Medline search. Here we report a case of live intravitreal adult Brugian filaria, where the parasite was successfully removed by pars plana vitrectomy. Identification of the worm was done by light microscopy and confirmed by immuno chromatographic test.

Estefanofilariose em humanos: ocorrência e mecanismos de transmissão / Stephanofilariasis in humans: occurrence and transmission mechanisms

Descreve-se aqui o encontro do helminto Stephanofilaria spp., em dermatite nodular ulcerativa em seres humanos, sendo as ulceras pesquisadas de dimensões variadas e com alguns anos de existência. Acredita-se que, a presença desse parasita, que afeta diversas espécies de animais domésticos e silvestres, possa agravar o quadro com as lesões produzidas nos vasos venosos e linfáticos, reduzindo a circulação, retardando a cicatrização. O ciclo deste parasita no ser humano, também é descrito.

This work describes the occurrence of the helminth Stephanofilaria spp in ulcerative nodular dermatitis in humans. The ulcers observed presented a variety of sizes and some years of existence. It is believed that the presence of this parasite, which affects several domestic and wild animal species, may harm human health through lesions produced in venous and lymphatic vessels that reduce blood circulation and retard the healing process. The cycle of this parasite in humans is also described.

The observation of microfilarial rate and density in cats inoculated with increasing numbers of Brugia pahangi infective larvae.

Having close kinship to Brugia malayi, B. pahangi is a member of the family Filariidae, which causes lymphatic filariasis in dogs and cats. Although this nematode is unlikely to cause a zoonotic disease in humans, study of the B. pahangi life cycle may help control human filariasis. The objective of this study was to examine microfilarial rates and densities of B. pahangi in experimentally induced infections in cats as a relative measurement. Cats were infected with 3 different amounts of 3rd-stage larvae (L3); 100, 300 and 500. Cats infected with 100 L3 became patent for microfilariae longer than the other groups (mean100 = 99+/-44 days). In comparison, the pre-patent period of B. pahangi was somewhat shorter in cats with 300 and 500 L3 infections (mean300 = 76+/-13 and mean500 = 63+/-5 days). The microfilarial densities of these cats were also determined; the density of microfilariae (mf/1 ml blood) increased relative to the duration of infection. One-way ANOVA tests were used to compare the microfilarial densities of the cats with varying numbers of L3. We found that the microfilarial density of cats with 500 L3 exhibited significant differences (p < 0.05) from cats with 300 and 100 L3. However, we concluded that the amount of microfilariae produced in the blood circulation of these cats were not increasing relative to the numbers of L3 taken by the host.

A juvenile filarial worm, Wuchereria bancrofti, extracted from the vitreous of the eye: the first report in the world literature.

A live worm was extracted from the vitreous of the eye of a patient in Sri Lanka. Based on the details of its length, width, oesophagus and vulva it was identified as Ajuvenile female Wuchereria bancrofti.

Culex quinquefasciatus in Phitsanulok as a possible vector of nocturnally periodic Wuchereria bancrofti transmission in Myanmar immigrants.

The present study was undertaken in order to study whether Culex quinquefasciatus collected in Phitsanulok Province can be an insect host for the development of Wuchereria bancrofti larvae. W. bancrofti infected blood from Myanmar workers in Mae Sot, Tak Province was fed to mosquitoes by using the artificial membrane feeding. An infection of W. bancrofti was found with the highest density of L3 in the mosquito thorax on the 14th day after feeding. The infection rate also correlated to the density of microfilaria found in the donor's blood. Our results showed that Cx. quinquefasciatus in Phitsanulok is a possible vector of nocturnally periodic W. bancrofti.

Transmission of filarial infection through blood transfusion.

Microfilariae can be transmitted by blood transfusion and they may be circulated in the recipient's blood but they do not develop into adult worms. Mortality associated with transfusion associated filarial infection is not documented but it may give rise to morbidity in transfusion recipients in terms of allergic reaction. The present study was carried out to investigate the association of post transfusion reactions and filarial infections in an endemic area. About 11,752 transfusion recipients were followed up and in 15 months period, 47 (0.4%) post transfusion reactions (PTR) were reported. Routine investigations for post transfusion reaction were carried out in all 47 patients and their respective blood donor. Moreover, blood culture, microfilaria detection by concentration technique, filarial antibody and antigen detection (both by ELISA) were done in all subjects. Out of 47 patients showing post transfusion reaction, 29 (61.7%) patients developed allergic reaction. Eighteen (38.3%) patients having allergic reaction did not have previous history of blood transfusion and 14 (29.8%) of them received transfusion from blood donors who was either positive for microfilaria, filarial antigen or antibody. Microfilaremia was demonstrated in 4 (8.5%) patients and 5 (10.6%) blood donors. Microfilaria was concurrently present in 2 patients and their respective donors. Filarial antibody was detected in 27 (56.5%) patients and 26 (55.3%) blood donors but microfilaria was detected in 3 (6.4%) and 4 (8.5%) subjects, respectively. Antigen detection test correlated with microfileraemic state of subjects. The result shows that transfusion associated filarial infection may be a probable cause for transfusion-associated morbidity in endemic areas. In 14 (29.8%) patients having allergic reactions, the probable cause was transfusion-associated filarial infection. Filarial antigen detection test was found to be more useful in detecting infections. Blood donors with active history of filarial infection should be deferred from donating blood. Filarial antigen detection test may be employed as screening test for blood donors, if possible.

A morfologia do intestino médio de Culex quinquefasciatus Say, 1823 e sua interação experimental com microfilárias de Litomosoides chagasfilhoi Moraes Neto, Lanfredi e De Souza, 1997 e Wuchereria bancrofti (Cobbold, 1877) Seurat, 1921 / The morphology of the midgut of Culex quinquefasciatus Say, 1823 and its interaction with experimental microfilariae of Litomosoides chagasfilhoi Moraes Neto, Lanfredi and De Souza, 1997 and Wuchereria bancrofti (Cobbold, 1877) Seurat, 1921

Analisamos a morfologia do intestino médio do mosquito Culex quiquefasciatus de região endêmica e a sua interação com as microfilárias de Wuchereria bancrofti (agente da filariose humana) e de Litomosoides chagasfilhoi (agente de filariose de roedores). O intestino médio está dividido em regiões torácica e abdominal como visto em outras espécies de mosquitos. Porém, o intestino médio torácico é formado por dois tipos celulares: células principais e basais; e o intestino médio abdominal por três tipos: células principais clara e basais. Existe uma rede muscular organizada revestindo externamente o órgão, a qual sofre mudanças irreversíveis no intestino médio abdominal após o repasto sangüíneo. A estrutura da matriz peritrófica e a sua formação induzida pela alimentação sangüínea são distintas das demais espécies de mosquitos. O L. chagasfilhoi invade o intestino médio entre 1h e 3h após o repasto infectivo, enquanto que a W. bancrofti entre 30 minutos e 5 horas com o maior número de invasão nas primeiras 2 horas. O L. chagasfilhoi preferencialmente invade o intestino médio abdominal enquanto que a W. bancrofti invade o intestino médio torácico, principalmente perto da região abdominal. Ambas espécies de microfilárias cruzam o intestino médio através do espaço intercelular, causando hipertrofia e extrusão de uma a duas células epiteliais. A perda da bainha pelas microfilárias de ambas espécies não é um pré-requisito para a evasão do intestino. Etapas subseqüentes do ciclo da W. bancrofti, tais como, a melanização na hemocele e a invasão precoce da musculatura de vôo foram também observadas. Não foi possível observar tais etapas nos mosquitos infectados com L. chagasfilhoi porque eles morreram poucas horas após a invasão do intestino médio. Estas diferenças no processo de interação das espécies de microfilárias provavelmente estão relacionados com a capacidade vetorial do mosquito.

A morfologia do intestino médio de Culex quinquefasciatus Say, 1823 e sua interação experimental com microfilárias de Litomosoides chagasfilhoi Moraes Neto, Lanfredi e De Souza, 1997 e Wuchereria bancrofti (Cobbold, 1877) Seurat, 1921

Analisamos a morfologia do intestino médio do mosquito Culex quiquefasciatus de região endêmica e a sua interação com as microfilárias de Wuchereria bancrofti (agente da filariose humana) e de Litomosoides chagasfilhoi (agente de filariose de roedores). O intestino médio está dividido em regiões torácica e abdominal como visto em outras espécies de mosquitos. Porém, o intestino médio torácico é formado por dois tipos celulares: células principais e basais; e o intestino médio abdominal por três tipos: células principais clara e basais. Existe uma rede muscular organizar revestindo externamente o órgão, a qual sofre mudanças irreversíveis no intestino médio abdominal após o repasto sangüíneo. A estrutura da matriz peritrófica e a sua formação induzida pela alimentação sangüínea são distintas das demais espécies de mosquitos. O L. chagasfilhoi invade o intestino médio entre 1h e 3h após o repasto infectivo, enquanto que a W. bancrofti entre 30 minutos e 5 horas com o maior número de invasão nas primeiras 2 horas. O L. chagasfilhoi preferencialmente invade o intestino médio abdominal enquanto que a W. bancrofti invade o intestino médio torácico, principalmente perto da região abdominal. Ambas espécies de microfilárias cruzam o intestino médio através do espaço intercelular, causando hipertrofia e extrusão de uma a duas células epiteliais. A perda da bainha pelas microfilárias de ambas espécies não é um pré-requisito para a evasão do intestino. Etapas subseqüentes do ciclo da W. bancrofti, tais como, a melanização na hemocele e a invasão precoce da musculatura de vôo foram também observadas. Não foi possível observar tais etapas nos mosquitos infectados com L. chagasfilhoi porque eles morreram poucas horas após a invasão do intestino médio. Estas diferenças no processo de interação das espécies de microfilárias provavelmente estão relacionados com a capacidade vetorial do mosquito

Subconjunctival adult bancroftian filarial worm.

We report a case of live adult Bancroftian Filarial nematode removed from the sub conjunctival tissue of a 58-year-old woman from the temporal limbus of her left eye.

Brugia malayi in a naturally infected cat from Narathiwat Province, southern Thailand.

Brugia malayi-like from an infected cat from Narathiwat Province, southern Thailand was identified intensively by microfilarial morphometry, acid phosphatase activity, and adult morphology. The results indicated that both microfilarial and adult characteristics conformed to the topotypic B. malayi.

Transmission efficiency of Culex quinquefasciatus and Aedes aegypti to Wuchereria bancrofti infection: an experimental study.

Present study was undertaken to evaluate the suitability of natural (Culex quinquefasciatus) and experimental (Aedes aegypti) vectors for supporting the development of W. bancrofti larvae for onward transmission. Both the species permitted development of W. bancrofti mf to infective larvae (L3) within 11 to 13 days. The mf intake by both the species of mosquitoes was directly related to mf density in donor's blood. Culex exhibited higher L3 recovery than Aedes. In Aedes maximum percent L3 development occurred after ingesting 4 mf whereas Culex exhibited best establishment at an average mf intake of 11.5. Nevertheless wide variation in mf density in donor's blood did not significantly affect the larval establishment in Aedes mosquito while in Culex very high (> 400 mf/40 microliters) or low (< 50 mf/40 microliters) mf counts in donor's blood adversely affected the L3 recovery. The results reveals that A. aegypti has an edge over the natural vector, Culex in being a voracious feeder, their easy laboratory maintenance and better transmission potential.

Breast filariasis--a case report.

An unusual presentation of filariasis as a breast lump simulating breast carcinoma, in a 50 year old woman residing in Pokhara, Nepal. The case was reported on Fine Needle Aspiration Cytology and also evaluated by histopathology. Morphology of the nematode is that of Brugia species which is unusual in Nepal.

Autogenous Aedes togoi sub-colony (Chanthaburi, Thailand strain), an efficient laboratory vector in study of filariasis.

Comparative filarial susceptibility and biology between stock colony and selectively autogenous Aedes. togoi sub-colony were carried out to determine the laboratorial vector-capacity and viability of autogenous sub-colony. The results of susceptibility revealed that the selectively autogenous Ae. togoi sub-colony yielded higher susceptibility than the stock colony, ie Dirofilaria immitis: susceptibility rates=80.00% [Exp1(F8)] and 76.19% [Exp2(F17)] (autogenous sub-colony), 53.33% (Exp1) and 71.43% (Exp2) (stock colony); Brugia malayi: susceptibility rates=83.33% [Exp1(F17)] and 84.38% [Exp2(F19)] (autogenous sub-colony), 81.25% (Exp1) and 75.00% (Exp2) (stock colony), but not at the level of statistically significant differences except the Exp1 of D. immitis, which was significant difference. In addition, the average No. L3 per infected mosquito in the selective autogenous sub-colony (D. immitis: Exp1=3.37, Exp2=3.19; B. mallayi: Exp1=8.80, Exp2=3.37) was also higher than in stock colony (D. immitis: Exp1=2.44, Exp2=2.73; B. malayi: Exp1=7.85, Exp2=3.02), but not at the level of statistically significant difference. The results of comparisons on some biological aspects demonstrated that most of the cases they have similar biology except the average egg deposition per gravid female of stock colony (130.17+/-43.33) was significantly more than selectively autogenous sub-colony(F9) (94.33+/-13.69), egg length x width 575.62+/-18.06 microm x 186.15+/-9.35 microm of stock colony was significantly larger than selectively autogenous sub-colony(F9) 560.49+/-18.96 microm x 177.99+/-8.40 microm, and mean longevity of adult female of stock colony [41.60(6-61)] was significantly longer than selectively autogenous sub-colony(F9) [35.00(5-39)]. The selectively autogenous sub-colony was established and twenty-two successive generations have been colonized.

Temporal variations in biting density and rhythm of Culex quinquefasciatus in tea agro-ecosystem of Assam, India.

Temporal changes in the biting density and host-seeking periodicity of Culex quinquefasciatus, the vector of bancroftian filariasis, were studied for two years in the tea agro-ecosystem of Assam, India. Average biting density of the vector varied from 0.8/hour in December to 13.3/hour in March. Minimum temperature was found to have a limiting effect on the biting density of the vector mosquitos. Multiple regression analysis showed that the rainfall and minimum temperature were significant factors influencing biting density of this vector mosquito. The biting rhythm of Cx. quinquefasciatus was found to be nocturnal with two distinct peak periods of biting activity. The first peak was around 19.00 to 20.00 hours and the second peak period was around 22.00 to 23.00 hours. Biting activity however was seen throughout the night with declining trend as the night proceeded.

Dietilcarbamazina no tratamento da filariose bancroftiana / Diethylcarbamazine in the treatment of Bancroft's filariasis

Os autores realizaram uma ampla revisão sobre o tratamento da filariose bancroftiana com a droga dietilcarbamazina. Os aspectos interessantes sobre o histórico de sua descoberta e os conceitos básicos de sua farmacologia foram relatados de forma resumida. Ênfase especial, por outro lado, foi dada às especulações feitas pelos diversos autores sobre os achados intrigantes descritos na literatura. Foram trazidos os novos avanços sobre o conhecimento da doença, como por exemplo, a visualização pela ultra-sonografia do verme vivo de Wuchereria bancrofti, no seu hospedeiro natural, o homem. Isso possibilitou a compreensão de muitos dos achados aparentemente paradoxais encontrados na literatura sobre o tratamento da infeção com a DEC. Assim, devido à inexistência de uma droga sucessora que reunisse efeitos micro e macrofilaricidas ideais e aos novos conhecimentos sobre a bancroftose e sobre a própria dietilcarbamazina, foi-lhe conferido um novo realce. Esses aspectos a colocaram numa posição de destaque no cenário da infecção, à época do seu quase cinqüentenário de existência.

The authors presented a detailed review about the treatment of bancroftian filariasis with diethylcarbamazine. The interesting aspects about the drug discovery and the basic concepts about its pharmacology were reported in a summarised form. On the other hand, emphasis was made about the speculation done by several authors about the intriguing findings regarding its efficacy reported in the literature. Latter, it was brought the new advances about the disease, as for example, the visualization by ultrasound of living Wuchereria bancrofti adult worm on its natural host--the human being. This made possible the comprehension of several paradoxical issues reported, focusing the treatment of infection using diethylcarbamazine. So far, because of the lack of ideal drug with micro and macrofilaricidal properties, together with the new understand about the disease and the new parameters for monitoring the efficacy of the drug, diethylcarbamazine has back its importance conquered at the begin of its discovery, almost fifth years ago.

Studies on the transmission potential of surviving microfilaremias after basic control of filariasis.

After filariasis was basically controlled (the microfilarial rate was lower than 1%) in Henan Province in 1987, longitudinal observation of the disease has been carried out in all the province in order to study the regular pattern of growth and decline or the transmission potential of the disease. According to the distribution of filaria species and original microfilarial rate, 7 administrative villages in 7 counties were selected as surveillance sites. From 1988 to 1995, etiological and mosquito vector surveys were made continuously in all sites where no control measure was conducted. 10 surviving microfilaremic individuals became negative gradually over the first 6 years and no new microfilaremia was found. Since then, the microfilarial rate was zero. During the 8 years, 19 vector mosquitos were positive, with a total of 33 filarial larva. Culex pipiens pallens was the predominant mosquito species inside human dwelling in all sites. The man-biting rate of mosquitos for outdoor sleepers fluctuated greatly, the highest was 360.60 mosquitos per man per night and the lowest 7.20. The man-biting rate of mosquitos for sleepers inside mosquito-nets was approximately 1. The proportion of multiparous mosquitos also fluctuated greatly, the highest was 88.10% and the lowest 27.27%. According to the data described above, the man-biting rate of mosquitos which contained filaria L3 was less than 1 mosquito per man per transmission season. It is suggested that after the microfilarial rate was lower than 1%, the surviving microfilaremias became negative gradually in 3-5 years, and the transmission of the disease was blocked. Therefore, in the districts where filariasis was basically controlled, elimination of the disease was attainable.