Larval Migration Research Articles

Susceptibility of entomopathogenic nematodes to ivermectin and thiabendazole

The objective of the present study was to determine the susceptibility of entomopathogenic nematodes to ivermectin and thiabendazole. Soil samples collected from the municipalities of Irapuato and León, Guanajuato, Mexico, were obtained, from which the entomopathogenic nematodes (EPNs) of the Steinernematidae and Heterorhabditidae families were isolated. The samples were classified from livestock and nonlivestock soils, and the susceptibility of EPNs to anthelmintics was determined with the larval motility assay (LMA, 24 h) and the larval migration inhibition assay (LMI assay, 48 h). Sterile distilled water (T1) and treatments with 1% ivermectin diluted in 5% DMSO (dimethyl sulfoxide) (T2) and 5% thiabendazole diluted in 5% DMSO (T3) were applied to infective juvenile larvae. Analysis of variance was performed with a factorial design and Tukey’s test at 0.05 probability. In addition, different concentrations of ivermectin (0.1, 0.5, 1, 1.2, 1.5, and 2 μg) and thiabendazole (1, 5, 10, 12, 15, and 20 mg) were evaluated to perform a Probit analysis to determine their LC50. All strains of EPNs were susceptible to ivermectin in both the LMA and LMI assay. The results show that EPNs are susceptible to ivermectin and thiabendazole, and the degree depends on the type of test performed, the chemical product used, and the origin of the strain of EPN.

S100A8/S100A9 deficiency increases neutrophil activation and protective immune responses against invading infective L3 larvae of the filarial nematode Litomosoides sigmodontis.

Neutrophils are essentially involved in protective immune responses against invading infective larvae of filarial nematodes. The present study investigated the impact of S100A8/S100A9 on protective immune responses against the rodent filarial nematode Litomosoides sigmodontis. S100A9 forms with S100A8 the heterodimer calprotectin, which is expressed by circulating neutrophils and monocytes and mitigates or amplifies tissue damage as well as inflammation depending on the immune environment. Mice deficient for S100A8/A9 had a significantly reduced worm burden in comparison to wildtype (WT) animals 12 days after infection (dpi) with infective L3 larvae, either by the vector or subcutaneous inoculation, the latter suggesting that circumventing natural immune responses within the epidermis and dermis do not alter the phenotype. Nevertheless, upon intradermal injection of L3 larvae, increased total numbers of neutrophils, eosinophils and macrophages were observed within the skin of S100A8/A9-/- mice. Furthermore, upon infection the bronchoalveolar and thoracic cavity lavage of S100A8/A9-/- mice showed increased concentrations of CXCL-1, CXCL-2, CXCL-5, as well as elastase in comparison to the WT controls. Neutrophils from S100A8/A9-/- mice exhibited an increased in vitro activation and reduced L3 larval motility more effectively in vitro compared to WT neutrophils. The depletion of neutrophils from S100A8/A9-/- mice prior to L. sigmodontis infection until 5dpi abrogated the protective effect and led to an increased worm burden, indicating that neutrophils mediate enhanced protective immune responses against invading L3 larvae in S100A8/A9-/- mice. Interestingly, complete circumvention of protective immune responses in the skin and the lymphatics by intravenous injection of L3 larvae reversed the phenotype and resulted in an increased worm burden in S100A8/A9-/- mice. In summary, our results reveal that lack of S100A8/S100A9 triggers L3-induced inflammatory responses, increasing chemokine levels, granulocyte recruitment as well as neutrophil activation and therefore impairs larval migration and susceptibility for filarial infection.

Effects of Gnathostoma spinigerum infective stage larva excretory-secretory products on NK cells in peripheral blood mononuclear cell culture: focused on expressions of IFN-γ and killer cell lectin-like receptors.

Human gnathostomiasis is mainly caused by third-stage larvae of Gnathostoma spinigerum (G. spinigerum L3). Excretory-secretory products (ES) released from infective helminthic larvae are associated with larval migration and host immunity modulation. Natural killer (NK) cells have important immune functions against helminth infection. Currently, the effects of ES from G. spinigerum L3 (G. spinigerum ES) on NK cell activity are unclear. This study investigated whether G. spinigerum ES affected human NK cells. Human normal peripheral blood mononuclear cell (PBMC) cultures were used to mimic immune cells within the circulation. PBMC were co-cultured with G. spinigerum ES (0.01-0.05μg/ml) for 5 or 7days. Levels of IFN-γ in cultured supernatants were measured by enzyme-linked immunosorbent assay. The expressions of mRNA encoding NK cell receptors, especially the C type killer cell lectin-like family (KLR; NKG2A, NKG2C, and NKG2D) and IFN-γ in ES induced PBMC were determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). ES induced PBMC markedly decreased the levels of IFN-γ and increased the expressions of NKG2A and NKG2D on NK cells. In conclusion, low amounts of G. spinigerum ES modulated NK cells by downregulating the transcription of IFN-γ and upregulating the expressions of KLR (NKG2A and NKG2D receptors) during the 7-day observation period. These findings indicate more in-depth studies of NK cell function are required to better understand the mechanism involved in immune evasive strategies of human gnathostomiasis.

Optimal age of Trichostrongylus colubriformis larvae (L3) for the in vitro larval exsheathment inhibition test under tropical conditions

This study identified the optimal age of Trichostrongylus colubriformis larvae (L3) under tropical conditions for the in vitro evaluation of plant extracts using the larval exsheathment inhibition test (LEIT). Two T. colubriformis isolates with different anthelmintic (AH) susceptibility status were used for this study. The L3 of both isolates were maintained on refrigeration (4−5 °C) until use. For the LEIT, the isolates were tested every week during 16 weeks, using a stock solution of Acacia pennatula acetone:water extract at different dilutions (80–1200 μg/mL). Respective positive controls (levamisole 12.5 mM) and negative controls (PBS) were included. Effective concentrations 50 % (EC50), 90 % (EC90) and respective 95 % confidence intervals (95 %CI) for exsheathment inhibition were calculated at 60 min after exposure to chlorine solution. Motility of L3 (migration percentages (M%)) was recorded weekly using the larval migration test (LMT) as an indicator of L3 fitness over time. No correlation was found between L3 age or M% and the extract’s EC50 or EC90 values. However, the EC50 values for the A. pennatula extract ranged from 80 to 200 μg/mL from weeks 2–10. Beyond week 12, larval exsheathment was irregular, with higher EC50 and EC90 values and wider 95 %CI. The M% decreased below 85 % on week 7 for Paraiso isolate, and on week 10 for FMVZ isolate. A linear negative relationship was observed between the age of L3 and M% for both T. colubriformis isolates. The relationship (slope) for both isolates was similar therefore a single linear equation was estimated describing all M% data (r2 = 0.771, df = 164, P < 0.05). Thus, when using these T. colubriformis isolates under our tropical conditions for the evaluation of AH activity of plant extracts with LEIT, the optimal age of L3 is between weeks 2–7, when M% remained above 85 %. The latter may ensure consistent and reproducible exsheathment results for T. colubriformis. Each laboratory must identify optimal conditions to perform the LEIT.

In vitro anthelmintic activity of Psidium guajava hydroalcoholic extract against gastro-intestinal sheep nematodes.

Tanniferous plants have been used for ruminants verminosis control and represent a possibility to minimize the pharmacological resistance against conventional antiparasitics. This study aimed to evaluate the antihelminthic activity of the hydroalcoholic extract of stem bark of guava tree (PgHA). It was performed the hatchability and larval migration inhibition assays to evaluate PgHA at the following concentrations 0.62, 1.25, 2.5 and 5.0 mg mL-1 and the control treatments. The total polyphenol, flavonoid and tannin contents were determined by phytochemical analysis, high performance liquid chromatography coupled to mass spectrometry. The antioxidant activity was evaluated by 1,1-diphenyl-2-picrylhydrazyl, ferric reducing antioxidant power and thiobarbituric acid reactive substances tests. It was also determinated total protein, intracellular H2O2 and antioxidant activity of enzimes: glutathione S-transferase and superoxide dismutase. PgHA was able to inhibit both hatchability and larval migration, but only hatchability inhibition presented dose-dependent pattern. The antioxidant activity was demonstrated by linear regression with IC50 corresponding to 534.02 μg mL-1. The antiparasitic mechanism occurred through pro-oxidative activity by the increase of total proteins, intracellular H2O2 and the lipid peroxidation products, as well as the increase of the enzymes above related. Thus, the PgHA showed antiparasitic activity in vitro.

Pathology of larvae and adults in dogs and cats.

Endoparasitism remains a significant cause of morbidity and mortality in puppies and kittens. The ascarids of dogs and cats (Toxocara canis, Toxascaris leonina, Toxocara cati) cause significant pathology due to either larval migration or adult nematode burden within the gastrointestinal tract. This chapter will review the important pathologic events and lesions that are encountered during ascarid development in dogs and cats with emphasis given to the characteristic gross pathology lesions that are seen in affected animals.

Acute fasciolosis outbreak with aberrant larval migration among sheep in Kashmir Valley

The present study describes the outbreak of acute fasciolosis among sheep in the twin districts of Budgam and Kupwara of the Kashmir Valley in 2019-20. To ascertain the cause of deaths, clinical examination and fecal examination of diseased sheep and post-mortem examination of the dead animals was carried out. The diseased sheep presented symptoms of cold extremities, anorexia, dullness, aloofness, depression, green-colored profuse watery discharge from nose and mouth (rumen contents) and recumbency before death. The liver showed typical migratory tracts due to fasciolosis. Fasciola spp. parasites were observed in liver and gall bladder during postmortem examination. The aberrant migration of the parasite was also noted in some cases with adult parasites recovered from lungs in only one case. All fecal samples tested negative for Fasciola spp. eggs. Most of the deaths were observed in pregnant ewes and younger sheep. Further mortality was controlled by mass dosing with Triclabendazole. The results indicated Acute Fasciolosis as the cause of death in these sheep as revealed by gross lesions and retrieval of adult Fasciola spp. from liver and gallbladder. The parasite demands serious attention due to its ill effects on animal health, economy of farmers and zoonotic implications.

Ascaris lumbricoidesandAscaris suumvary in their larval burden in a mouse model

Ascariasis is a neglected tropical disease, caused by Ascaris lumbricoides, affecting 800 million people worldwide. Studies focused on the early stage of parasite infection, occurring in the gut, liver and lungs, require the use of a mouse model. In these models, the porcine ascarid, Ascaris suum, is often used. The results obtained from these studies are then used to draw conclusions about A. lumbricoides infections in humans. In the present study, we sought to compare larval migration of A. suum and A. lumbricoides in mouse models. We used a previously developed mouse model of ascariasis, which consists of two mouse strains, where one mouse strain - C57BL/6J - is a model for relative susceptibility and the other - CBA/Ca - for relative resistance. Mice of both strains were infected with either A. suum or A. lumbricoides. The larval burden was assessed in two key organs, the liver and lungs, starting at 6 h post infection (p.i.) and ending on day 8 p.i. Additionally, we measured the larval size of each species (μm) at days 6, 7 and 8 p.i. in the lungs. We found that larval burden in the liver is significantly higher for A. lumbricoides than for A. suum. However, the inverse is true in the lungs. Additionally, our results showed a reduced larval size for A. lumbricoides compared to A. suum.

Nematocidal Effect of Oyster Culinary-Medicinal Mushroom Pleurotus ostreatus (Agaricomycetes) against Haemonchus contortus.

The nematocidal effect of Pleurotus ostreatus (white variety of oyster mushroom) aqueous extract (AE) was evaluated against Haemonchus contortus eggs and infective larvae (L3) in vitro and in artificially infected gerbils (Meriones unguiculatus). The chemical analyses indicated that constituents of AE are tridecanoic, tetradecanoic, linolelaidic, 9,15-octadecadienoic, and oxalic acids. P. ostreatus extract inhibited larval hatching by 100% at the concentration of 2.24 mg/mL and (50% effective concentration) EC50 of 0.73 mg/mL. In the larval development test, AE induced a larvicidal effect at the concentration of 50 mg/mL and EC50 of 17.24 mg/mL. The larval migration test revealed a reduction of 94.7% at a concentration of as low as 4 mg/mL and EC50 of 1.25 mg/mL. No significant effects of treatment with P. ostreatus AE were seen on H. contortus in the gerbil model. Thus, our results demonstrate an important nematocidal in vitro effect of P. ostreatus AE against the parasite H. contortus. However, further investigations are necessary to confirm the anthelmintic potential of P. ostreatus extract in small ruminants.

Species of Anisakidae nematodes and Clinostomum spp. infecting lisa Mugil curema (Mugilidae) intended for human consumption in Mexico.

Anisakisspp. nematodes are potentially zoonotic parasites; that infects a wide variety of aquatic species worldwide, with marine fish being the paratenic hosts. The aim of study was identify the presence of Anisakidae nematodes, and other parasites in Mugil curema . A total of 96 M . curema obtained from local markets in Tulancingo, Hidalgo, Mexico, were analyzed by necropsy. Only five M . curema present nematode collection in epaxial muscle. The tissues with the highest prevalence of parasites were identified, and samples of epaxial muscle with larval migration analyzed by histopathology. Visible parasites in necropsy tissues were classified according to their morphology. Nematode found in the liver were Contracaecum spp. (41.17%) and Pseudoterranova spp. third stage (7.36%); in the caudal part of the kidney were Anisakis spp. (13.23%), Pseudoterranova spp. third stage (11.77%) and Contracaecum spp. (5.88%); and in epaxial muscle were Anisakis spp. Larva I (5.88%) and Pseudoterranova spp (4.42%). In one fish, Clinostomum spp. was detected in epaxial caudal muscle. The present work reports for the first time the presence of nematodes of the family Anisakidae and Clinostonum spp. metacercariae, with zoonotic potential, in M . curema intended for human consumption in Tulancingo, Hidalgo, Mexico.

Larval Migration Behaviour of Busseola fusca (Lepidoptera: Noctuidae) on Bt and Non-Bt Maize under Semi-Field and Field Conditions.

Busseola fusca (Fuller) (Lepidoptera: Noctuidae) is a destructive pest of maize throughout the African continent. Bt maize is an effective control measure for this pest, however, selection pressure for resistance evolution is high. This necessitates the implementation of insect resistance management (IRM) strategies such as the high-dose/refuge strategy. This IRM strategy relies on the validity of several assumptions about the behaviour of pests during insect-hostplant interactions. In this study, the migration behaviour of B. fusca larvae was evaluated in a semi-field (greenhouse) and field setting. The effect of factors such as different Cry proteins, plant growth stage at infestation, and plant density on the rate and distance of larval migration were investigated over four and five week periods. Migration of the larvae were recorded by using both a leaf feeding damage rating scale and destructive sampling at the end of the trials. Results indicated that B. fusca larval migration success was significantly affected by plant growth stage and plant density—while limited larval migration was recorded in plots inoculated with larvae at a late growth stage (V10), higher plant density facilitated increased interplant migration. The results also suggest that B. fusca larvae do not migrate extensively (rarely further than two plants from the natal plant) and that larval mortality is high. Implications for IRM strategies are discussed.

Effect of essential oils on cattle gastrointestinal nematodes assessed by egg hatch, larval migration and mortality testing.

The efficacy of eight essential oils (EOs) (Solidago canadensis, Eucalyptus globulus, Pelargonium asperum, Ocimum basilicum, Thymus vulgaris, Mentha piperita, Cymbopogon citratus and Cymbopogon martinii) against gastrointestinal nematodes (GINs) was evaluated using eggs collected from naturally infected cattle and cultured infective larvae (L3). The larvae species cultured from the faecal samples and subjected to two in vitro tests were Haemonchus spp. (55.5%), Trichostrongylus spp. (28.0%), Cooperia spp. (15.0%) and Oesophagostomum spp. (1.5%). The genus of EO Cymbopogon (C. citratus and C. martinii) showed the highest anthelmintic activity at the dose of 8.75 mg/ml, for the egg hatch, the larval migration and mortality assays. All of the EOs tested reduced egg hatching to rates <19.0%, compared to the controls (water and water + Tween 20) that had rates >92.0%. Cymbopogon citratus and C. martinii treatments resulted in 11.6 and 8.1% egg hatch, had the lowest migration of larvae through sieves, 60.5 and 54.9%, and the highest mortality rates, 63.3 and 56.3%, respectively. Dose-response tests showed that EO from C. citratus had the lowest larval LC50 and migration inhibition concentration (IC50) values of 3.89 and 7.19 mg/ml, respectively, compared to two other EOs (C. martinii and O. basilicum). The results suggest that EOs from the genus Cymbopogon can be interesting candidates for nematode control in cattle, although it may prove challenging to deliver concentrations to the gastrointestinal tract sufficient to effectively manage GINs.

Ovicidal and larvicidal potential of Rosmarinus officinalis to control gastrointestinal nematodes of sheep.

Gastrointestinal Nematode Infection (GIN) are the main constraint to the production of small ruminants. Studies of medicinal plants have been an important alternative in the effort to control these parasites. Therefore, the purpose of this study was to evaluate the in vitro ovicidal and larvicidal activity of essential oil of Rosmarinus officinalis. The oil was extracted, analyzed by gas chromatography and tested on GIN eggs and larvae in six concentrations, 227.5mg/mL, 113.7mg/mL, 56.8mg/mL, 28.4mg/mL, 14.2mg/mL and 7.1mg/mL. To determine the ovicidal activity, GIN eggs were recovered from sheep feces and incubated for 48h with different concentrations of the oil. For the evaluation of larval migration, third-stage larvae (L3) were obtained by fecal culture, and associated with the essential oil for 24h at the same concentrations, after which they were left for another 24 hours on microsieves, followed by the count of migrating and non-migrating larvae. The assays of R. officinalis oil showed a significant (p<0.05) 97.4% to 100% inhibition of egg hatching and a significant (p<0.05) 20% to 74% inhibition of larval migration. The main constituent revealed by gas chromatography was Eucalyptol. The results indicate that R. officinalis essential oil has ovicidal and larvicidal activity on sheep GINs.

Combining data from different sampling methods to study the development of an alien crabChionoecetes opilioinvasion in the remote and pristine Arctic Kara Sea

Data obtained using three different types of sampling gear is compared and combined to assess the size composition and density of a non-indigenous snow crab population Chionoecetes opilio in the previously free of alien species Kara Sea benthos. The Sigsbee trawl has small mesh and catches even recently settled crabs. The large bottom trawl is able to catch large crabs, but does not retain younger crabs, due to its large mesh. Video sampling allows the observation of larger crabs, although some smaller crabs can also be spotted. The combined use of such gear could provide full scope data of the existing size groups in a population. The density of the crabs was calculated from the video footage. The highest figures were in Blagopoluchiya Bay at 0.87 crabs/m2, where the settlement seems to be reaching its first peak of population growth after the introduction. High density in the Kara Gates Strait at 0.55 crabs/m2, could be due to the close proximity of the Barents Sea from where the crabs can enter by both larval dispersal and active adult migration. All size groups have been present in most sampled areas, which suggest successful settlement and growth of crabs over a number of years. Again, this was not the case in Blagopoluchiya Bay with high density of small crabs (<30 mm CW), which confirms its recent population growth. Male to female ratio was strikingly different between the bays of the Novaya Zemlya Archipelago and west of the Yamal Peninsula (0.8 and 3.8 respectively). Seventy five ovigerous females were caught in 2016, which confirms the presence of a reproducing population in the Kara Sea. The spatial structure of the snow crab population in the Kara Sea is still in the process of formation. The presented data indicates that this process may lead to a complex system, which is based on local recruitment and transport of larvae from the Barents Sea and across the western Kara shelf; formation of nursery grounds; active migration of adults and their concentration in the areas of the shelf with appropriate feeding conditions.

Combination of quercetin and ivermectin: In vitro and in vivo effects against Haemonchus contortus

The aim of the present study was to evaluate the in vitro effect of quercetin combined with ivermectin (IVM) on Haemonchus contortus larvae and adults with different resistance profiles and demonstrate the in vivo anthelmintic action of this combination when used in sheep naturally infected. The effect of combination was evaluated based on the analysis of the mean effective concentration (EC50) obtained for larvae using the larval migration inhibition test and for adults using the motility test on females. The tests with larvae and adults were conducted using isolates with different degrees of susceptibility to IVM (sensitive, intermediate and highly resistant). The in vivo effect was evaluated based on the reduction in the egg count (FEC) and reduction in the count of adult helminths recovered after parasitological necropsy. Using the combination of quercetin with IVM, it was observed that in larvae, quercetin did not significantly reduce the EC50 for IVM in the sensitive and highly resistant isolates, but led to a significant reduction in the EC50 for IVM in the intermediate isolate. In adults, quercetin did not significantly reduce the EC50 for IVM in any of the isolates. No significant effect of the combination was found regarding the reduction in FEC or total count of parasites. The results of the in vitro and in vivo tests performed in the present study on quercetin activity underscore the importance of evaluating resistance-reversing agents among different stages of parasite development as well as among isolates with different resistance profiles. The action of quercetin combined with IVM on the motility of H. contortus larvae and adults was influenced by the degree of resistance and development stage of the parasite. The combination was effective only on intermediate resistant larvae. No action of the combination against adults was found. Moreover, this combination, when administered through the intra-abomasal route, was not effective at reducing the FEC and parasite load of naturally infected sheep.

In vitro anthelmintic activity of the crude hydroalcoholic extract of Piper cubeba fruits and isolated natural products against gastrointestinal nematodes in sheep

This study describes the in vitro anthelmintic activity of a hydroalcoholic extract from the fruit of Piper cubeba and its major isolated components against the eggs and larvae of gastrointestinal nematodes obtained from naturally-infected ovines. In vitro anthelmintic activity was evaluated using the egg hatch test (EHT), larval development test (LDT) and L3 migration inhibition test (LMT). The extract showed ovicidal and larvicidal activity, with an EC50 of 200 μg/mL and 83.00 μg/mL in the EHT and LDT, respectively. The extract inhibited 100% of larval migration at the lowest tested concentration (95 μg/mL). The crude extract was purified using successive silica gel chromatographic columns, which revealed the lignans hinokinin, cubebin and dihydrocubebin as the major compounds that were present, which were then used in in vitro tests. Cubebin, dihydrocubebin and hinokinin showed higher activity than the crude extract, with an EC50 for ovicidal activity of 150.00 μg/mL, 186.70 μg/mL and 68.38 μg/mL, respectively. In the LDT, cubebin presented an EC50 of 14.89 μg/mL and dihydrocubebin of 30.75 μg/mL. Hinokinin inhibited 100% the larval development at all concentrations evaluated. In the LMT, dihydrocubebin inhibited 100% the larval migration in all concentrations evaluated while cubebin and hinokinin showed EC50 values of 0.89 μg/mL and 0.34 μg/mL, respectively. P. cubeba extract is rich in several classes of active compounds, but here we demonstrate that the described anthelmintic activity may be related to the presence of these lignans, which are present in larger concentrations than other components of the extract. Our results demonstrate for first time the anthelmintic activity against gastrointestinal nematodes in sheep for this class of special metabolites that are present in P. cubeba fruit. However, future detailed studies are needed to evaluate the effectiveness of P. cubeba fruits extract and active lignans in in vivo tests.

Downstream migration and mortality of larval Ayu (Plecoglossus altivelis altivelis) on the Sanriku coast, northern Japan

The ecology and mortality of larval ayu (Plecoglossus altivelis altivelis) during downstream migration was investigated during three spawning seasons (2008–2010) in the Unosumai River, Iwate Prefecture, Japan. Downstream larval migration occurred from early September to mid-December, peaking from late September to mid-October. Larval drift numbers recorded at the lowest reach spawning ground peaked from 18:00–21:00, twenty-four-hour-surveys in 2008 and 2010 resulting in total daily numbers of drifting larvae of 2,855,660 and 785,880, respectively, with 88.1 and 88.7%, respectively, occurring between 17:00–24:00. A twenty-four-hour-survey at the river mouth in 2010 recorded larvae between 19:00–06:00 (following day), peaking at 21:00 (67.5% of larvae occurred between 21:00–23:00), approximately 2 h later than the peak at the lowest reach spawning ground. The river mouth survey estimated a total larval number of 113,724, corresponding to 14.5% of the number recorded from the lowest reach spawning ground on the same day, indicating a mortality rate of 85.5%.

Histopathological characterization of Toxocara canis- and T. cati-induced neurotoxocarosis in the mouse model.

Infective larvae of Toxocara canis and T. cati, the common roundworms of dogs and cats, may invade the central nervous system of paratenic hosts, including humans, causing neurotoxocarosis (NT). Previous studies on NT in the model organism "mouse" have indicated distinct differences between T. canis and T. cati regarding larval migration patterns as well as the severity of clinical symptoms and behavioural alterations. The objective of the present study was to provide an extensive characterization of the underlying histopathological alterations, comparing T. canis- and T. cati-induced changes in different brain areas over the course of murine infection. Four histological sections of five brains each of T. canis- and T. cati-infected as well as uninfected C57Bl/6 mice were investigated 7, 14, 28, 42, 70 and 98days post infection (dpi), while brains of T. cati-infected and control mice were also available 120 and 150 dpi. In addition to haematoxylin-eosin and luxol fast blue-cresyl violet staining, immunohistochemistry was employed to study microglia/macrophage cell morphology and to detect accumulation of β-amyloid precursor protein (β-APP) as an indicator of axonal damage. Haemorrhages, eosinophilic vasculitis and activated microglia/macrophages were detected in both infection groups starting 7 dpi, followed by eosinophilic meningitis in cerebra as from 14 dpi. Overall, little differences in the proportion of animals affected by these alterations were found between the two infection groups. In contrast, the proportion of animals displaying β-APP accumulation was significantly higher in the T. canis than T. cati group as from 28 dpi regarding the cerebrum as well as at 98 dpi regarding the cerebellum. In T. canis-infected mice, myelinophagic microglia/macrophages ("gitter cells") appeared as from 14 dpi, whereas these were first observed at 70 dpi in T. cati-infected animals. The proportion of animals displaying demyelination and/or gitter cells in the cerebrum was significantly higher in the T. canis than T. cati group as from 28 dpi, and at 28 and 42 dpi regarding the cerebellum. Earlier and more severe neurodegeneration during T. canis- than T. cati-induced NT, especially in the cerebrum, may explain the differences in behavioural alterations observed in previous studies. In addition to differences in larval migration preferences, immunological processes may contribute to these patterns, which warrant further investigation.

Occurrence of Rat Lungworm (Angiostrongylus cantonensis) in Invasive Coqui Frogs (Eleutherodactylus coqui) and Other Hosts in Hawaii, USA

The rat lungworm (Angiostrongylus cantonensis) has emerged as an important human and animal health concern in Hawaii, US. Although the life cycle of the parasite requires both rat and gastropod hosts, other animals acting as paratenic hosts, such as frogs and centipedes, have been identified as sources of infection. We investigated the occurrence of rat lungworm infections in potential paratenic hosts in Hawaii to provide information on how they might be involved in transmission of angiostrongyliasis. We confirmed the presence of rat lungworm in 87% (21/24) of introduced Puerto Rican coqui frogs (Eleutherodactylus coqui) in Hilo, Hawaii, by real-time PCR. Additionally, four Cuban greenhouse frogs (Eleutherodactylus planirostris), two cane toads (Rhinella marina), and three centipedes (Scolopendra subspinipes) were found to be infected. In the frogs and toads, multiple tissue types were positive, including stomach and intestine, muscle, liver, heart, and brain, indicating larval migration. We identified rat lungworm infections in frogs, toads, and centipedes in Hawaii and highlighted the lack of knowledge of the role paratenic hosts may be playing in the transmission and life cycle maintenance of rat lungworm in Hawaii.

Lupin (Lupinus spp.) seeds exert anthelmintic activity associated with their alkaloid content

The growing range of drug resistant parasitic nematode populations threatens the sustainability of ruminant farming worldwide. In this context, nutraceuticals, animal feed that provides necessary dietary requirements while ensuring parasite control, could contribute to increase farming sustainability in developed and low resource settings. In this study, we evaluated the anthelmintic potential of lupin seed extracts against the major ruminant trichostrongylids, Haemonchus contortus and Teladorsagia circumcincta. In vitro observations showed that seed extracts from commercially available lupin varieties could significantly but moderately inhibit larval migration. This anthelmintic effect was mediated by the seed alkaloid content and was potent against both fully susceptible and multidrug resistant H. contortus isolates as well as a susceptible T. circumcincta isolate. Analytical chemistry revealed a set of four lupanine and sparteine-derivatives with anthelmintic activity, and electrophysiology assays on recombinant nematode acetylcholine receptors suggested an antagonistic mode of action for lupin alkaloids. An in vivo trial in H. contortus infected lupin-fed ewes and goats failed to demonstrate any direct anthelmintic effect of crude lupin seeds but infected lupin-fed goats suffered significantly less parasite-mediated blood losses. Altogether, our findings suggest that the anthelmintic potential of lupin remains limited. However, the potent alkaloids identified could lead to the development of novel drugs or may be used in combination with current anthelmintics to improve their efficacy.