Impact of single dose of diethylcarbamazine and other antifilarial drug combinations on bancroftian filarial infection variables: assessment after 2 years

The impact of single dose mass drug administration of diethylcarbamazine (DEC), DEC with albendazole (ALB), and ivermectin (IVR) with albendazole, was examined on the human bancroftian filarial infections in village scale trials in south India, from a follow-up study after 2 years. The treatment arms administered with DEC alone and DEC+ALB demonstrated long-term benefits in reducing microfilaraemia significantly (P<0.05), while antigenaemia reduction was negligible. The arm with ALB+IVR did not show such reductions. Among the antigenaemic and microfilaraemic individuals, 87% became amicrofilaraemic in DEC+ALB arm, which were higher than that observed in the other 2 treatment arms. Among amicrofilaraemics (but Ag+), nearly 35% cleared of infection in DEC+ALB, while 26% and 6% in DEC alone and IVR+ALB arms, respectively. The drug combination DEC+ALB was observed to demonstrate a significant impact in reducing filarial infection even after 2 years post treatment.     

Ivermectin inhibits molting of Wuchereria bancrofti third stage larvae in vitro

The effect of ivermectin or diethylcarbamazine (DEC) on Wuchereria bancrofti molting from the third to the fourth larval stage (L3 to L4) was evaluated in vitro. L3 larvae were harvested from laboratory-reared Aedes togoi 2 wk after feeding upon a microfilaremic human volunteer. The larvae were kept in an artificial medium (Franke's NI medium) with 10% human serum under an atmosphere of 5% CO2 for 20 days. Experimental tubes also contained ivermectin (0.1-1,000 ng/ml) or DEC (0.1-10,000 ng/ml). An estimated concentration of 50 ng/ml ivermectin inhibited molting in 50% of the larvae expected to molt. For DEC, this value was roughly 1,000 ng/ml. In this in vitro culture system, ivermectin inhibited the L3 to L4 molt of W. bancrofti and was roughly 20-fold more potent in this activity than DEC.     

Influence of anti-filarial chemotherapy strategies on the genetic structure of Wuchereria bancrofti populations

Lymphatic filarial (LF) parasites have been under anti-filarial drug pressure for more than half a century. Currently, annual mass drug administration (MDA) of diethylcarbamazine (DEC) or ivermectin in combination with albendazole (ALB) have been used globally to eliminate LF. Long-term chemotherapies exert significant pressure on the genetic structure of parasitic populations. We investigated the genetic variation among 210 Wuchereria bancrofti populations that were under three different chemotherapy strategies, namely MDA with DEC alone (group I, n = 74), MDA with DEC and ALB (group II, n = 60) and selective therapy (ST) with DEC (group III, n = 34) to understand the impact of these three drug regimens on the parasite genetic structure. Randomly amplified polymorphic DNA profiles were generated for the three groups of parasite populations; the gene diversity, gene flow and genetic distance values were determined and phylogenetic trees were constructed. Analysis of these parameters indicated that parasite populations under ST with a standard dose of DEC (group III) were genetically more diverse (0.2660) than parasite populations under MDA with DEC alone (group I, H = 0.2197) or with DEC + ALB (group II, H = 0.2317). These results indicate that the MDA may reduce the genetic diversity of W. bancrofti populations when compared to the genetic diversity of parasite populations under ST.     

Suppression of Brugia malayi (sub-periodic) larval development in Aedes aegypti (Liverpool strain) fed on blood of animals immunized with microfilariae

Preliminary studies were carried out to investigate the role of filarial specific antibodies, raised in an animal model against the filarial parasite, Brugia malayi (sub-periodic), in blocking their early development in an experimental mosquito host, Aedes aegypti (Liverpool strain). In order to generate filarial specific antibodies, Mongolian gerbils, Meriones unguiculatus, were immunized either with live microfilariae (mf) of B. malayi or their homogenate. Mf were harvested from the peritoneal cavity of Mongolian gerbils with patent infection of B. malayi and fed to A. aegypti along with the blood from immunized animals. Development of the parasite in infected mosquitoes was monitored until they reached infective stage larvae (L3). Fewer number of parasites developed to first stage (L1) and subsequently to L2 and L3 in mosquitoes fed with blood of immunized animals, when compared to those fed with blood of control animals. The results thus indicated that filarial parasite specific antibodies present in the blood of the immunized animals resulted in the reduction of number of larvae of B. malayi developing in the mosquito host.     

Brugia malayi: effects of antibacterial agents on larval viability and development in vitro

Recent studies have suggested that intracellular Wolbachia bacteria are necessary for reproduction and survival of adult filarial worms. We now report results of in vitro studies of effects of antibacterial antibiotics (tetracycline, rifampicin, chloramphenicol, azithromycin, and doxycycline) on Brugia malayi infective larvae (L3) motility and molting. All of the antibiotics tested except chloramphenicol decreased L3 motility by 50% or more at 10 days, with minimal effective concentrations (MECs) of 20-100 microg/ml. Tetracyclines, rifampicin, and chloramphenicol inhibited L3 to L4 molting by 12 days in a concentration- and time-dependent manner, with MECs in the range of 1-20 microg/ml. These studies show that antibiotics active against Rickettsiaceae inhibit B. malayi L3 molting at low concentrations in vitro; higher concentrations kill the larvae. While it is possible that antibiotics directly affect filarial L3, we believe it is more likely that the effects seen are indirect effects related to bacterial killing.     

Selection of different genotype larvae and adult worms for anthelmintic resistance by persistent and short-acting avermectin/milbemycins

To understand the factors that influence selection for anthelmintic resistance, it is necessary to examine the impact of drug treatment, particularly persistent drugs, on all phases of the worm life cycle. The efficacy of various avermectin/milbemycin anthelmintics was determined against resident worms, incoming larvae (L3) and development of eggs in faecal culture. Homozygote-resistant and maternal and paternal F1-heterozygote genotypes of Haemonchus contortus were used to infect sheep before or after treatment with ivermectin (IVM) oral, IVM capsule, moxidectin (MOX) oral or MOX injectable. Total worm count and quantitative larval culture were used to determine efficacy against parasitic and free-living stages, respectively. Selection for resistance by IVM capsules occurred at the adult and L3 stages because of poor efficacy against these stages for all resistant genotypes. However, the selective advantage of these surviving worms was reduced due to the low development of their eggs to L3 in faecal culture. For MOX, selection for resistance predominantly occurred after treatment because of high efficacy against resident adult worms of all resistant genotypes but poor efficacy against resistant L3 ingested after drug administration. The results indicated no evidence of sex-linked inheritance for IVM resistance. Mean IVM efficacies against homozygous and heterozygous resistant adult worms were not different, and IVM capsule efficacy against incoming L3 was approximately 70% for all resistant genotypes, consistent with a dominant trait. MOX was highly effective against adults of all resistant genotypes and approximately 76% effective against incoming L3 regardless of resistance genotype, also consistent with a dominant trait. These results will enable the impact of persistent drugs on worm control and anthelmintic resistance to be estimated. The results indicate that IVM capsules should not be used in populations where avermectin/milbemycin resistance is present.     

The cuticular proteins from free-living and parasitic stages of Haemonchus contortus--I. Isolation and partial characterization

1. Cuticles were isolated from the adult males, adult females, the second molt (2M) sheath from the infective larvae (L3(2M)), and the parasitic third stage (L3) of the sheep parasite Haemonchus contortus by a combination of mechanical disruption and detergent treatment. 2. The 2ME soluble cuticular proteins from adult males contained 4 or 5 major protein bands with molecular weights ranging from 100 to 56 kD with the most prominent band at 56 kD. The cuticular proteins from adult females were similar to the male. 3. Cuticular proteins from the larval stages, 2M cuticle, and L3 cuticle, differed from the adults and from each other. The most prominent protein bands were observed with molecular weights on 78 and 39 kD for the L3 cuticle and 100, 91 and 46 kD for the 2M cuticle. The 2ME soluble cuticular proteins from all developmental stages were at least partially digested by bacterial collagenase. 4. The amino acid composition of cuticular proteins was similar for the L3 and 2M, but adults had lesser amounts of glycine and greater amounts of basic amino acids than the larval stages. The amount of the isolated cuticle solubilized by the 2ME treatment was greatest in adults (80%) compared to the L3 (64%) and the 2M (22%). 5. These results support a hypothesis that there are quantitative and qualitative stage specific differences in the cuticular proteins of H. contortus.     

Possible direct effect of diethylcarbamazine on the infective larvae of Brugia pahangi

The direct action of diethylcarbamazine (DEC) on the infective larvae of Brugia pahangi was studied. The larvae were cultured in RPMI 1640 supplemented with foetal bovine serum and antibiotics for 22 days. Most of the larvae remained alive for 8 days, but survival rate of larvae decreased rapidly from day 10 onwards. The larvae did not grow in the culture system. The addition of DEC did not affect the morbidity of the larvae and no difference was observed in the morphological characteristics between the larvae cultured in the presence or absence of DEC. The infective larvae were cultured in vitro for 5 days in the presence or absence of DEC, and inoculated into jirds. The animals were necropsied at intervals, and developing larvae and adult worms were recovered. When the larvae were cultured without DEC and then inoculated subcutaneously into jirds, 29.8% of the inoculum was recovered 3-15 days, and 25% 19-22 weeks, post-inoculation. However, when the larvae were exposed to DEC in vitro and inoculated into jirds, the rate of recovery was reduced to 25% 3-15 days post-inoculation and 2% after 19-22 weeks. When the control larvae cultured in vitro were inoculated intraperitoneally into jirds, 41.3% of inoculum was recovered 3-15 days, and 42.8% 19-22 weeks, post-inoculation. Again the corresponding value for larvae exposed to DEC in vitro was reduced to 19.8% 3-15 days, and 8% 19-22 weeks, post-inoculation. It was observed that the larvae exposed to DEC in vitro were retarded in their development in jirds.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunity to Litomosoides carinii in Mastomys natalensis. II. Effects of chemotherapeutically abbreviated and postpatent primary infections on challenges with various stages of the parasite

Naive Mastomys natalensis, Litomosoides carinii-infected M. natalensis at a postpatent stage of the infection and L. carinii-infected M. natalensis treated chemotherapeutically with furazolidone (FUR), FUR and diethylcarbamazine (FUR/DEC) or amoscanate (AMOS) were challenged by either injection or implantation of 40 third stage larvae (L3, s.c.), 40 fourth stage larvae (L4, 16 days old, i.p.), 20 male and 20 female preadult worms (36 days old, i.p.), 12 adult female worms (i.p.) or 6 X 10(6) microfilariae/kg (i.v.). Microfilaraemia in animals challenged at a postpatent stage (independent of the kind of challenge), was either totally suppressed or at least greatly reduced. Necropsy of L3-challenged animals showed that neither the length of the worms nor their content of morphologically intact, intrauterine stages was affected. Infected, treated animals challenged with developing stages (L3, L4 and preadult worms) showed reduced levels of microfilaraemia (by up to 75%). Dissection of AMOS-treated, L3-challenged animals showed that both the developmental rate and the fertility of the worms were affected. Microfilaraemia was also reduced after implantation of adult worms into treated animals. This was independent of the interval between treatment and challenge (44-150 days) except in animals challenged 10 days after AMOS-treatment, which showed no difference from naive controls. However, infected, treated M. natalensis, cotton rats and gerbils did not develop immunity against intravenously injected blood microfilariae.     

Effects of diethylcarbamazine on the motility of Angiostrongylus cantonensis and Dirofilaria immitis

Effects of piperazine derivatives, especially of diethylcarbamazine (DEC) on adult Angiostrongylus cantonensis and Dirofilaria immitis were examined. Piperazine (3 X 10(-5)-10(-4) M) paralyzed A. cantonensis and the action was antagonized by picrotoxin. 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP) (10(-5)-10(-4) M) caused contraction but little effect was produced by strychnine. An inhibitory effect on untreated preparations was caused by lower concentrations (3 X 10(-6)-10(-5) M) of diethylcarbamazine citrate (DEC) and also on the preparations contracted by eserine. A stimulatory effect was also seen when higher concentrations (10(-4)-3 X 10(-4) M) of this drug were applied to both preparations. The inhibitory action of DEC was antagonized by gabergic antagonists such as picrotoxin and bicuculline, but not by alpha-adrenergic antagonists like dibenamine and phentolamine. When the worm preparation was paralyzed by strychnine or hexylresorcinol (inhibitors of the release of acetylcholine in this worm), the stimulatory effect of DEC was blocked, but pyrantel (a nicotinic cholinergic agonist) contracted the paralyzed preparation. However, the effect of DEC on D. immitis (10(-7)-3 X 10(-4) M) was inhibitory, and this action was also antagonized by picrotoxin. These results suggest that the DEC inhibitory and stimulatory action is through the gabergic and cholinergic mechanisms in adult A. cantonensis and D. immitis.     

Accumulation of copper by roots,hypocotyls, cotyledons and leaves of sunflower (Helianthus annuus L.)

The effects of different concentrations of copper sulfate on the growth of and the accumulation of Cu2+ by root, hypocotyl, cotyledon and leaf growth of sunflower (Helianthus annuus L.) were examined in this study. The concentrations of copper sulfate (CuSO4 x 5H2O) used were in the range from 10(-5) to 10(-3) M. Seedlings exposed to 10(-5) M Cu2+ solution exhibited a 33% increase in growth (P < 0.005) when compared with the root length of the control. The seedlings treated with 10(-3) M Cu2+ were significantly inhibited in shoot growth (P < 0.005). The Cu2+ content in roots, hypocotyls, cotyledons and leaves increased with increasing solution Cu2+ concentration. The roots of plants exposed to 10(-3) M Cu2+ accumulated a large amount of Cu (1070 microgram/g DW), and the Cu2+ level was approximately 25 fold higher than that of control. The Cu2+ contents in sunflower roots treated with 10(-4) and 10(-5) M Cu2+ were about 3.3 and 2.6 fold higher than the control, respectively. Also, the Cu2- level of the roots exposed to 10(-3) M Cu2+ was approximately 7.7 and 9.8 fold respectively, in comparison with the roots of plants grown in 10(-4) and 10(-5) M Cu2+. At 10(-3) M Cu2+, the Cu accumulated mainly in the roots (about 73%), and small amounts of Cu2+ (27%) were translocated to the hypocotyls, cotyledons and leaves. The Cu2+ concentration in the roots was less than that of the above parts of seedlings in treated groups with 10(-5) - 10(-4) M Cu2+. H. annuus has potential ability to accumulate Cu without being overly sensitive to Cu toxicity.     

CD8+ T lymphocytes are not required for murine resistance to human filarial parasites.

Mice are resistant to the establishment of infection with the nematode parasite Brugia malayi, an etiologic agent of human lymphatic filariasis. We have recently shown that T and B lymphocyte-deficient C.B.-17 scid/scid mice are permissive for infection with this parasite, whereas coisogenic C.B.-17+/+ mice are resistant. This observation suggests that T and B lymphocytes that comprise the antigen-specific immune system orchestrate murine resistance to B. malayi. In order to define the component of the antigen-specific immune response that is responsible for this resistance, we have tested the susceptibility of beta 2M-/- mice to infection with B. malayi L3 larvae. These mice are homozygous for insertional disruption of their B2m genes, which encode beta 2-microglobulin, the small subunit of the major histocompatibility (MHC) antigens. They do not express beta 2-microglobulin and, as a consequence, fail to express the class I major histocompatibility antigens, and they do not develop the CD8+ class I MHC-restricted cytotoxic T cell subset. We find that these mice are completely resistant to B. malayi, indicating that the CD8+ T lymphocyte subset is not an obligate requirement for murine resistance to human filarial parasites.     

Studies with Brugia pahangi 17. The anthelmintic effects of diethylcarbamazine.

Diethylcarbamazine (DEC) was active in vitro against infective larvae and microfilariae of Brugia pahangi but only at high concentrations. When fed to mosquitoes which were infected with B. pahangi it had little or no activity. In jirds it was inactive against B. pahangi microfilariae and adults when administered at 300 mg/kg for 5 days either by the intraperitoneal or oral route. In cats given 25 or 50 mg DEC/kg intraperitoneally on 3 or 5 occasions it was not microfilaricidal, but most of the adult worms died within 30 days of the end of treatment. Although most microfilariae disappeared from the blood of cats immediately (i.e., within an hour) after treatment, they reappeared within a few hours in the same numbers. Microfilarial levels were reduced after treatment but there was no precipitate decline as occurs in human B. malayi patients.     

Thyroid stimulating hormone enhancement of bovine oocyte maturation in vitro.

Efforts to improve bovine embryonic development in vitro involved study of effects of thyroid stimulating hormone (TSH) alone or in combination with LH on bovine oocyte maturation (IVM). Putative effects were assessed by observing cumulus expansion (CE), fertilization (IVF), and development to morulae/blastocysts (M/B). Effects of prolactin (PRL) were also investigated. Variables for the 24-hr IVM interval were no hormone (control), TSH (0.1, 0.5, or 1.0 micrograms/ml) or PRL (10, 100, or 1000 micrograms/ml), luteinizing hormone (LH) (0, 10, or 100 micrograms/ml) + TSH (0.1 or 0.5 micrograms/ml), and serum (20%, v/v) + 0.5 micrograms TSH/ml; data were from 4-5 trials for each IVM treatment. Higher proportions of oocytes exhibited complete CE with hormones or serum than without (P less than 0.05). All oocytes (with and without CE) were inseminated with heparin-capacitated sperm. A higher proportion of inseminated oocytes cleaved after IVM with 0.5 micrograms TSH/ml (53.4%) than for other TSH treatments (P less than 0.05). The combination of TSH (0.1 and 0.5 micrograms/ml) with 10 micrograms LH/ml for IVM enabled higher proportions (P less than 0.05) of ova to fertilize (67.4 and 69.2%) than did medium alone (28.3%), LH (10 micrograms/ml) alone (54.1%) or serum + 0.5 micrograms TSH/ml (55.6%). No improvement in proportions undergoing fertilization was seen after addition of TSH to 100 micrograms LH/ml for IVM. Frequency of CE and cleavage did not differ among PRL treatments. More M/B developed from cleaved ova after IVM with LH or TSH than with PRL or no hormone (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

与中红侧沟茧蜂生境与寄主定位相关的玉米及棉铃虫幼虫体表挥发性成分的提取与鉴定

【目的】鉴定玉米Zea mays L.和棉铃虫Helicoverpa armigera (Hübner)幼虫体表挥发性成分中对中红侧沟茧蜂 Microplitis mediator (Haliday)具有生境与寄主定位作用的信息化合物,从化学生态学的角度研究玉米-棉铃虫-中红侧沟茧蜂三重营养关系,解释中红侧沟茧蜂寻找寄主的过程中的信息识别机制,为害虫的综合防治的“推-拉”方法提供一定的理论基础。【方法】利用触角电位仪(EAG)、触角电位联用仪(GC-EAD)、气质联用仪(GC-MS)及“Y”型嗅觉仪确定玉米和棉铃虫幼虫体表提取物的信息化合物。在室内利用玉米以及棉铃虫幼虫体表挥发物标准品化合物以及模拟混合物,使用“Y”型嗅觉仪进行中红侧沟茧蜂成虫行为反应试验。【结果】玉米挥发物中有11种化学成分,棉铃虫幼虫体表挥发物中有6种化学成分对中红侧沟茧蜂的触角具有电生理活性,其中4种成分在两种挥发物中都存在。室内行为反应试验发现：与正己烷对照相比,玉米的模拟组分对雌、雄蜂均表现出显著（P<0.05）的诱引作用;棉铃虫1龄幼虫体表模拟组分对雌蜂具有极显著的诱引作用（P<0.01）,对雄蜂具有显著的诱引作用（P<0.05）;棉铃虫2龄幼虫体表模拟组分对雌蜂具有显著的诱引作用（P<0.05）。【结论】本研究证明了玉米以及棉铃虫幼虫体表挥发物中分别存在11种（庚醛、2-己醇、1-己醇、1-辛烯-3-醇、壬醛、葵醛、苯甲醛、反式-2-壬烯-1-醇、己酸、苯基乙醇、月桂醇）和6种（2-己醇、己酸乙酯、1-己醇、壬醛、辛酸乙酯、癸醛）中红侧沟茧蜂生境及寄主定位的化学信息物质。     

Detection of resistance to ivermectin in Haemonchus contortus.

Infective, third-stage (L3) larvae of Haemonchus contortus isolates resistant to ivermectin (IVM) show a decreased sensitivity to IVM-induced paralysis in vitro. The inhibition of larval motility by IVM can be detected in L3 larvae incubated in the dark on an agar matrix containing IVM, by the failure of affected larvae to move when stimulated by exposure to light. Optimally, avermectin (AVM) potency is quantified after three cycles, each involving storage in the dark for 24 h followed by a brief exposure to light. For IVM-susceptible isolates, a 50% inhibition of motility (LP50) was achieved with IVM concentrations between 0.30 and 0.49 microM, while LP50 values in IVM-resistant isolates ranged from 0.8 to 2.6 microM depending on the in vivo resistance status of the isolate. A limited study of structure-activity relationships within the AVM class indicated that in vitro inhibition of L3 motility was consistent with the known in vivo efficacy of each analogue. Resistance factors for IVM-resistant isolates were dependent on AVM structure with the more polar AVM B2 analogue being a particularly sensitive probe of IVM-resistance status.     

Development of the nematode eyeworm, Thelazia skrjabini (Nematoda: Thelazioidea), in experimentally infected face flies, Musca autumnalis (Diptera: Muscidae)

The development of Thelazia skrjabini Erschow, 1928, was studied in experimentally infected laboratory-reared Musca autumnalis De Geer. Thelazia skrjabini developed to the infective third stage in a minimum of 9 days in M. autumnalis maintained at 27 +/- 2 C. First-stage larvae were not observed postinoculation, but second-stage larvae were first observed 3 days postinoculation. Development was asynchronous. Second- and third-stage larvae occur in capsules, occasionally in the head but primarily in the abdomen attached to fat bodies. First-stage larvae have anteriorly 1 ventral and 2 dorsal hooks, directed posteriorly. Second-stage larvae have 4 submedian cephalic papillae and faint annular striations. Third-stage larvae have 6 labial papillae, 4 submedian cephalic papillae and pronounced annulations. Morphometric studies of each larval stage were performed with specimens in glycerine.     

The effect of diethylcarbamazine on microfilariae of Litomosoides carinii in vitro and in vivo

Culture-derived Litomosoides carinii microfilariae (MFF) were used in in vitro and in vivo systems to investigate the effect of diethylcarbamazine (DEC) on these MFF. In vivo: Male rats, Mastomys natalensis, all of the same age, were injected intrathoracically (12) or intraperitoneally (36) with 10(3) or 10(4) MFF. After 30 min one half of each group of rats was given DEC per os. At 30, 60, and 120 min after DEC administration, two rats from the treated and two from the untreated group were bled and killed. The pleural or peritoneal cavities were rinsed with warm saline (0.15 M NaCl) to recover MFF. In both the intrathoracic and intraperitoneal experiments, equal numbers of MFF were recovered from treated and control rats at 30 and 120 min. However, at 60 min 85.5% fewer were recovered from the treated than from the nontreated animals. MFF were not found in the blood. In vitro: MFF were added to tissue culture dish wells (Linbro Div., Flow Labs, Hamden, Conn) prepared as follows: DEC-Serum (serum from normal rats given DEC at 500 mg/kg), DEC + Serum (serum with added DEC), serum only, RPMI 1640 only, and RPMI 1640 + DEC. Furthermore, the five treatments were prepared either with or without unstimulated peritoneal exudate (PE) cells. At 30 min in the DEC-Serum wells 45% of the MFF had adherent PE cells; in the remaining wells these cells adhered to 11% or fewer MFF. We interpret the aforementioned phenomena as representing the first step in the trapping and elimination of MFF after DEC treatment of L. carinii-infected M. natalensis.     

Pleiotropic effects of the neuropeptides CCAP and myosuppressin in the beetle, Tenebrio molitor L.

Biological activities of crustacean cardioactive peptide (CCAP; PFCNAFTGCa) and leucomyosuppressin (Lem-MS; pQDVDHVFLRFa) were studied in heterologous bioassays in the larvae and adults of Tenebrio molitor. CCAP exerted a reversible and dose-dependent cardio-stimulatory effect on the semi-isolated heart of the experimental beetles at a concentration of >10(-7) M and induced an effect similar to the endogenic cardio-stimulatory peptide, proctolin. Injections of CCAP (10(-9)-10(-3) M) into 4-day-old adult reproductive females increased the concentration of soluble proteins in hemolymph in comparison to the saline injected controls. Electrophoretic analyses indicated significant increase in the level of two proteins 130 and 170 kDa, and a partial increase of the level of 67-kDa protein. The studies indicated that CCAP increased also free hemolymph sugar concentration in young larvae and adults of the mealworm beetle. The cardio-inhibitory peptide Lem-MS exerted the opposite effect: at concentration 10(-7)-10(-6) M, it significantly decreased the heartbeat frequency. The induced changes were dose-dependent and reversible, but at higher concentrations (>10(-5) M) the stimulatory effect disappeared. Injections of the Lem-MS into young larvae at concentrations 10(-9)-10(-3) M, also increased the free hemolymph sugar level similarly to the CCAP. This work demonstrates the pleiotropic effects of CCAP and Lem-MS in Tenebrio molitor.     

Onchocerca volvulus, O. gutturosa, Brugia malayi, and Dirofilaria immitis: a comparative study of the immunochemical properties of cuticular proteins from filarial parasites

We compared the chemical and immunological properties of cuticular collagens from four species of filarial nematodes, Onchocerca volvulus, O. gutturosa, Brugia malayi, and Dirofilaria immitis. The electrophoretic mobility of the major polypeptides extracted from adult worms is characteristic for each species studied. Cuticular collagens from adult worms and infective larvae differ in their susceptibility to proteases that cleave vertebrate collagens and to collagenases prepared from different developmental stages of filarial parasites. The overall amino acid composition of filarial collagens resembles that of vertebrate interstitial collagens and differs from that reported for collagens from free-living or intestinal nematodes. However, cuticular proteins of the four filarial species studied significantly differed in amino acid composition and in their reactivity with antisera to interstitial and basement membrane collagens of vertebrates.