Worm Length Research Articles

Goats are more susceptible to Haemonchus contortus infection than sheep under similar experimental settings

Due to differences in their feeding behavior, sheep and goats are often assumed to respond differently to helminth infections. The present study compared Haemonchus contortus infection profile between sheep and goats under the same experimental setting. Experimental infection was conducted using a randomized block design in four groups of intact sheep (InfSH and ConSH) and goats (InfG, and ConG). Groups InfSH and InfG (N = 7 each) received 10,000L3 of H. contortus whereas the control groups ConSH and ConG (N = 7 each) remained uninfected. Faecal egg counts and PCV were measured from Day 0 to day 56 post infection (PI). On day 56 PI, animals were humanely slaughtered and abomasal contents were recovered to measure worm burden, worm length and in utero egg count. The findings show that: (1) Haemonchus infected animals showed an increase in FEC starting from day 21 PI, (2) progressive reduction in PCV was registered from day 7 PI and continued to the end of the experiment while this remained at pre-infection levels in control groups, (3) FEC was much higher (P < 0.001) and PCV was significantly lower (P < 0.05) in infected goats than in infected sheep, (4) at necropsy, total worm burden with worm establishment rates of 63% and 28.87% were registered respectively for infected goats and sheep with significant difference (P < 0.05), (5) Female worms were significantly longer (P < 0.05) in InfG (22.8(± 1.2) compared to InfSH (20.5 ± 0.67 mm) while (6) Mean worm fecundity was 974.8 ± 239.4 and 1162.5 ± 89.4 respectively for groups InfSH and InfG with no significant difference (P = 0.07), and (7) Parasite traits such as worm burden, FEC and female worm length were well correlated in sheep whereas few such patterns were observed in goats. In conclusion, under the same experimental infection, Arsi-Bale goats are more susceptible to H. contortus infection than Arsi-Bale sheep and hence deserve special attention when they are forced to live on grazing rather than browsing.

Secretory Cells in Halla parthenopeia (Oenonidae): Potential Implications for the Feeding and Defence Strategies of a Carnivorous Burrowing Polychaete.

Carnivorous polychaetes are known to bear diversified and often unique anatomical and behavioural adaptations for predation and defence. Halla parthenopeia, a species known to be a specialized predator of clams, thrives in the soft bottoms of the Mediterranean Sea, holding potential for polyculture and biotechnology due to the secretion of bioactive compounds. Our objective was to provide a comprehensive description of H. parthenopeia's anatomy and microanatomy, shedding light on the relation between morphology and habitat, chemical defences, and feeding behaviour. The pharynx, housing maxillae and mandibles connected to an extensive mucus gland, occupies a considerable portion of the worm's length, reaching beyond the oesophagus. This unique gland is responsible for secreting the feeding mucus, which immobilizes and aids in the digestion of clams probably acting as a vehicle of bioactive compounds synthesized by specialized serous cells in the mouth. Moreover, H. parthenopeia combines behavioural tactics, such as burrowing, and anatomical defences to evade predators. Examination of its epidermis revealed a thick cuticle layer and abundant mucocytes secreting locomotion mucus, both of which save the worm from mechanical harm during movement. When it is preyed upon, the worm can release a substantial amount of Hallachrome, a toxic anthraquinone produced by specific cells in its distal region. This pigment, with its known antimicrobial properties, likely acts as a chemical shield in case of injury. The results suggest that the ability of H. parthenopeia to prey on bivalves and to provide mechanical protection plus defence against pathogens rely on its ability to secrete distinct types of mucus. The interplay between highly specialized microanatomical features and complex behaviours underscores its adaptation as a predator in marine benthic environments.

Cross infection of Haemonchus contortus between calves and lambs

Haemonchus contortus can frequently be found infecting pre-weaned beef calves on sheep and beef farms around the North Island of New Zealand. The purpose of this study was to determine whether parasites cycling in young cattle constitute a potentially important source of infection for sheep.A field isolate of H. contortus was cycled through either calves or lambs for 3 generations. The larvae resulting from the third cycle of infection were then used to infect both lambs and calves and the resulting faecal nematode egg count (FEC), worm burden, adult worm length and in utero egg count were measured.Larvae derived from lambs inoculated into calves exhibited lower establishment rates, the adult worms were shorter, had lower in utero egg counts, and the resulting faecal egg counts were also lower than when inoculated into lambs (p < 0.01).H. contortus' lack of ability to passage freely between lambs and calves indicates that large populations are unlikely to occur under mixed grazing, resulting in limited potential as a source of infection in sheep. However, indications of an ability to adapt to the alternative host suggest that some investigation of infection in cattle dominant farming operations in the north of the country might be warranted.

Combined effects of Limonene and Ivermectin on P-glycoprotein-9 gene expression of lambs Infected with Haemonchus contortus

Although ivermectin (IVM) has a wide spectrum and long half-life, its frequent use as an anthelmintic for the last 42 years led to its worldwide tolerance by Haemonchus contortus. We evaluated the combination of limonene (LIM), a P-glycoprotein (Pgp) modulator, with IVM in lambs infected with a multidrug-resistant H. contortus. Twenty-four male Dorper lambs were artificially infected with two doses (seven days apart) of 8000 infective larvae of a multidrug-resistant isolate of H. contortus. The infection was patent 25 days later. Fifteen days before treatment with IVM (DAY –15), animals were divided into 4 groups: Infected-untreated control (CTL), IVM, LIM, and LIM+IVM. From DAY –15 to DAY + 14, groups LIM and LIM+IVM received 200 mg/kg of body weight/day of LIM via oral. On DAY 0, a single dose of IVM at 200 µg/kg of body weight was administered orally to groups IVM and LIM+IVM. On DAY + 7 and DAY + 14, fecal egg counts (FEC) were performed and on DAY + 14 animals were euthanized for total worm count (TWC), worm length, fecundity of females, and Pgp-9 gene expression. On DAY + 7, group LIM+IVM had 96.29% efficacy based on Fecal Egg Count Reduction TEST (FECRT) and a highly significant reduction in FEC (P = 0.0005) when compared to CTL. On DAY + 14, the efficacy of LIM+IVM was 82.87% on FECRT, although no differences were found among groups for FEC, TWC, worm length, or Pgp-9 gene expression. Female worms from the CTL group had higher egg counts in their uterus when compared to LIM. No differences were found for hematological or biochemical parameters, body weight, or weight gain among groups. Thus, LIM given daily at 200 mg/kg was safe for animals and, when combined with IVM, decreased egg shedding and could reduce pasture contamination, although it was unable to kill multidrug-resistant H. contortus.

Growth and Reproduction Rate of Eisenia fetida, using Lagerstroemia speciosa and Waste Disposable Face Masks to Test the NPK Level of the Compost

The main objective of the study is to estimate the NPK produced in the waste disposable of a Face Mask (FM) is decomposed with a medicinal plant Lagerstroemia speciosa (LS). The experimental design consists of 5 groups; each group contains yard soil, cow dung, and coir pith as compost bedding materials. T1 - control, T2 - KWCO, T3 - FMCO, T4 - LSCO and T5 - ESLS. The result shows that the growth rate of Eisenia fetida depends on the length and weight of worms. The reproduction rate varies depending on compost materials, with higher rates in T4 and lower rates in T1. The T5 shows a high level but is a little lower than the T4. Juvenile numbers are low in T2 and T3 trials. NPK Level: The nitrogen level 413.2±2.90** is highest in T4 and T3 is 404.3±1.78. Low in T1 336.6±2.84 trial, moderate in 376.3±3.13** T2 trial and T5 consist of 372.3±2.59**. The Phosphorus is commendably high in T5 1011.3±2.59** and T1 contains 992.3±3.06. The moderate amount is in T4 894.3±2.22**. The lowest level of P 623±2.94** is in T2 and T3 contain 676.6±2.88**. The potassium amount is varying in each trial. T1 is the most prominent amount of Potassium, and the level of K is 1900.3±1.18. The kitchen waste T2 trial produced 1840.6±2.37** and the amount of K is in 1670.3±1.18** T4. The most important face mask blue and black colour samples is mixed with kitchen waste T3 produces 1700±3.77** and medicinal Plant T5 which produces 1569.3±2.84**. Our conclusion is that the medicinal plants had a significant effect on the growth and reproductive rate of red wrigglers, and this effect was positively correlated with the NPK level in the soil.

Effect of sire fecal egg count estimated breeding value on parasite resistance traits in Haemonchus contortus infected Katahdin lambs

In the midst of anthelmintic resistance, genetic resistance to gastrointestinal nematodes (GIN) in lambs could reduce anthelmintic requirements; therefore, the fecal egg count (FEC) estimated breeding value (EBV) was developed as a measure of genetic merit for parasite burden. One of the first breeds to effectively implement the FEC EBV was Katahdin. To better understand the relationship between the FEC EBV and Haemonchus contortus (Hc) challenge infection, a divergent mating scheme was established with extremely high (High FEC, n = 5) or extremely low (Low FEC, n = 5) FEC EBV Katahdin rams over 2 years. Purebred lambs sired by these rams were born beginning in mid-March and managed on pasture until approximately 120 days of age. A primary infection was established based on FEC during this period. At this point, lambs (n = 109 in Year 1, n = 114 in Year 2) were removed from pasture, treated with an anthelmintic to reduce FEC and transported to the Animal Sciences Farm at West Virginia University. After a rest period, lambs were given 10,000 (Year 1) or 5000 (Year 2) Hc L3. Body weights, FEC, and packed cell volume were collected weekly. Lambs were harvested (n = 60/year) at 5 weeks post-infection. Abomasum worm counts were determined, and worm length was measured using Image J. Statistical analysis was performed by year using the Mixed Model procedure of SAS (SAS Institute, Cary, NC) with fixed effects of sire EBV type. Change in FEC after the prepatent period was greater in Year 1 High FEC-sired lambs compared to Low FEC-sired lambs (210 vs. 34 eggs/g, respectively; P < 0.05). At harvest in Year 2, a greater proportion of Low FEC- than High FEC-sired lambs had worm counts of zero (P < 0.05). Worm fecundity was lower in lambs sired by Low FEC rams (P < 0.05). Taken together, sire selection for low FEC EBV will lower FEC and worm count and improve GIN resistance in progeny.

Effects of Zinc Accumulation on Earthworm Aporrectodea caliginosa (Haplotaxida: Lumbricidae)

Earthworms are key to the earth's ecosystem, which helps the soil increase its fertility and repair its existing elements, as well as remove contaminants. This study investigated the accumulation of Zn in the earthworm Aporrectodea caliginosa after 15 days of exposure. The worms were grown in media with different concentrations of Zn between 750 and 1500 ppm. Each treatment consisted of three replicates, containing 30 worms. A control group without Zn was also used. Data were analyzed by using Duncan Multi–Range. The results revealed that A. caliginosa had a strong ability to accumulate Zn in its tissue compared to the control group. It was noticed that the increase of the heavy metal in the worm´s tissue is associated with the elevation of the metal in its media. The results show a significant loss of weight in the worm´s body and loss in the growth rate; they also shown a decrease in specific growth rate. Moreover, there was a significant decrease in the worm length, showing a high effect, especially after 15 days of breeding with all concentrations used in this study. The researchers recommend using earthworms to purify the soil from contaminants because earthworm has a great ability to get rid of all pollutants, whether metals or pesticides or parasites, especially in industrialized countries and agricultural lands.

Saturated C22-tailed cationic surfactant in concentrated brine: Structural evolution of wormlike micelles and rheological properties

Saturated C22-tailed ultra-long-chain imparts cationic surfactant stable chemical structure and strong hydrophobic interaction that favors the generation of wormlike micelles in brine. Such viscoelastic wormlike micellar fluid is expected to exhibit excellent salt-resistance and temperature-resistance. In this work, the self-assembled microstructures and microscopic flow properties of docosyl(trimethyl)azanium chloride (DCTAC) NaCl solutions were systematically investigated by means of cryogenic transmission electron microscopy (cryo-TEM), rheo-small angle neutron scattering (rheo-SANS), steady-state and dynamic rheology. It is found that DCTAC forms long, linear or branched, flexible or stiff wormlike micelles depending on NaCl concentration. The increase in salt content simultaneously promotes the growth of worm length and the formation of branches. While the worm chain elongation plays a dominant role in mid-range of NaCl concentrations, the branching formation acts as a leading role in supper high NaCl concentrations, which results in rheological properties experiencing a significant increase followed by a certain decrease upon increasing NaCl content up to ∼ 20 wt%. Moreover, DCTAC shows much better thickening power than corresponding homologue surfactants bearing shorter hydrophobic tail in brine, and DCTAC solution also shows much better temperature-resistance than them. These findings are helpful to understand salt effect on cationic surfactant self-assemble behavior, and the relationship between microstructure and solution macroscopical properties.

Praziquantel promotes protection against Schistosoma japonicum infection in mice

Praziquantel (PZQ) is the first line drug for the treatment of schistosomiasis. Several studies have confirmed that PZQ regulates host immunity, and we have recently found that pretreatment with PZQ enhances resistance against Schistosoma japonicum infection in buffaloes. We speculate that PZQ induces physiological changes in mice that prevent S. japonicum infection. To test this hypothesis and provide a practical measure to prevent S. japonicum infection, we determined the effective dose (the minimum dose), protection period and onset time of protection by comparing the worm burden, female worm burden and egg burden in PZQ-pretreated mice and blank control mice. Morphological differences between parasites were observed by measuring the total worm length, oral sucker, ventral sucker and ovary. The levels of cytokines, nitrogen monoxide (NO), 5-hydroxytryptamine (5-HT) and specific antibodies were measured using kits or soluble worm antigens. Hematological indicators on day 0 were analyzed in mice that received PZQ on days -15, -18, -19, -20, -21 and -22. The PZQ concentrations in plasma and blood cells were monitored using high performance liquid chromatography (HPLC). The effective dose was found to be two oral administrations (interval of 24 h) at 300 mg/kg body weight (BW) or one injection at 200 mg/kg BW, and the protection period of PZQ injection was 18 days. The optimal preventive effect was observed at two days post-administration, with a >92% worm reduction rate and significant worm reduction until 21 days after administration. Adult worms from PZQ-pretreated mice were runtish showing a shorter length, smaller organs and fewer eggs in the uteri of females. Detection of cytokines, NO, 5-HT and hematological indicators showed that PZQ induced immune-physiological changes, including higher levels of NO, IFN-γ and IL-2, and a lower level of TGF-β. No significant difference in the anti-S. japonicum specific antibody levels was observed. The PZQ concentrations in plasma and blood cells 8 and 15 days post-administration were lower than the detection limit. Our results confirmed that pretreatment with PZQ promotes the protection of mice against S. japonicum infection within 18 days. Although we observed some immune-physiological changes in the PZQ-pretreated mice, the exact mechanisms involved in the preventive effect require further study.

Antibody response and abomasal histopathology of lambs with haemonchosis during supplementation with medicinal plants and organic selenium

We evaluated the effects of dietary supplementation with medicinal plants (Herbmix) or organic selenium (Selplex) on the immune responses and histopathology of lambs infected with Haemonchus contortus. Twenty-seven lambs were infected and reinfected with approximately 11,000 third-stage larvae of H. contortus during the experiment (on days 0, 49 and 77). Lambs were divided into two supplemented experimental groups (Herbmix and Selplex) and unsupplemented group (Control). The abomasal worm counts at necropsy on day 119 were lower for Herbmix (4230) and Selplex group (3220) than to the Control (6613) which resulted in 51.3% and 36.0% of reduction, respectively. The mean length of adult female worms was in the order Control > Herbmix > Selplex (2.1, 2.08, and 2.01 cm, respectively). The specific IgG response against adults was significantly affected by time (P < 0.001). Serum-specific and total mucus levels of IgA in the Herbmix group were highest on day 15. Mean levels of serum IgM against adults were influenced by treatment (P = 0.048) and time (P < 0.001). The Herbmix group had strong local inflammation in the abomasal tissue, with the formation of lymphoid aggregates and the infiltration of immune cells, but the tissues of the Selplex group had higher numbers of eosinophils, globule leukocytes, and plasma cells. The lymph nodes of each animal had reactive follicular hyperplasia due to the infection. Dietary nutritional supplementation with a mixture of medicinal plants or organic selenium could improve local immune responses and thus enhance the resistance of animals to this parasitic infection.

Effect of three feeding levels on the pathogenesis and establishment of Haemonchus contortus in parasite-naïve Pelibuey hair sheep lambs during their first infection

This study evaluated the effect of three feeding levels on the pathogenesis and establishment of H. contortus upon the first infection of parasite-naïve Pelibuey hair sheep lambs. Forty-two 6-month-old hair sheep lambs (24 ± 4 kg) raised parasite free from birth were used. The lambs were assigned to 3 groups (n = 14), and each was fed a diet designed for different daily weight gain (DWG): 75 g/d (Diet 1), 125 g/d (Diet 2) and 200 g/d (Diet 3). After four weeks of diet adaptation, 10 lambs/group were infected with 450 L3H. contortus/kg BW (infected), and 4 lambs/group were kept parasite-free (NInf). DWG, hematocrit (Ht), hemoglobin (Hb), peripheral eosinophils (EOS), IgG concentration against H. contortus, and eggs per gram (EPG) of feces were measured in each lamb from day 14 before infection until day 29 postinfection (PI). On day 29 PI, the lambs were slaughtered to determine the total number of adult parasites (TAW), the length of the female worms, and the number of eggs in utero (EIU). Each group reached the expected DWG (P = 0.001), and there was no effect of infection or the diet × infection interaction. Ht was lower in infected lambs than in NInf lambs, and this difference was significant for animals on Diets 1 and 2 (P = 0.044). From day 14 PI onward, Hb was lower in the infected lambs than in the NInf lambs (P = 0.001). Furthermore, compared with NInf lambs, the infected lambs had higher EOS from day 7 PI and higher IgG from day 14 PI. Neither EOS nor IgG were affected by diet. Lambs on Diet 3 had lower EPG during patency than those fed Diets 1 or 2 (days 25 and 28 PI; P = 0.002). Furthermore, lambs fed Diet 3 had lower TAW (Diet 1 vs 3 P = 0.037; Diet 2 vs 3 P = 0.049) and EIU (P = 0.004) than lambs fed Diet 1 or 2. Lambs were resilient to infection regardless of diet. Although EOS and IgG were higher in all infected animals than in Ninf animals, EPG, TAW and EIU decreased only in lambs fed Diet 3. Thus, a diet targeting a DWG of 200 g/d can significantly limit the establishment of H. contortus in Pelibuey lambs infected for the first time.

Ascaridia galli eggs obtained from fresh excreta, worm uteri or worms cultured in artificial media differ in embryonation capacity and infectivity

Ascaridia galli infection models use eggs isolated from chicken excreta, worm uteri and worms cultured in artificial media. The aim of this study was to compare the infectivity of A. galli eggs isolated from these sources under two infection regimens. A 3 × 2 factorial arrangement was employed to test the infectivity of A. galli eggs from the three sources and two modes of infection (single or trickle infection). One hundred and fifty-six Isa-Brown one day-old cockerels randomly assigned to the six treatment groups (n = 26) were orally infected with embryonated A. galli eggs obtained from the three A. galli egg sources (worm uteri, excreta or eggs shed in vitro) administered either as single dose of 300 eggs at one day-old or trickle infected with 3 doses of 100 eggs over the first week of life. Twenty-two negative control birds remained uninfected. Eggs obtained from cultured worms or excreta exhibited a higher embryonation capacity (P = 0.003) than eggs obtained from worm uteri. There were higher worm establishment (infectivity) rates from embryonated eggs originating from cultured worms and worm uteri compared with eggs obtained from fresh excreta (P < 0.0001). Trickle infection resulted in a significantly higher total worm burden (P = 0.002), establishment rate (P = 0.002) and excreta egg counts (EEC, P = 0.025) than single infection. Worm length was greater in birds infected with embryonated eggs from excreta than from uteri or cultured worms (P < 0.0001). However, mode of infection did not affect worm length (P = 0.719) and weight (P = 0.945). A strong significant positive linear correlation was observed between EECs and female worm counts at 12 weeks of post infection sampling (r = 0.75; P < 0.0001). Body weight of birds was negatively correlated with both worm burden (r = − 0.21; P < 0.01) and EEC (r = − 0.20; P < 0.05) at 12 weeks post infection. In conclusion, our results show that eggs shed by cultured worms or isolated from worm uteri had greater infective capacity than eggs harvested from excreta and that trickle rather than bolus infection resulted in higher worm establishment. These factors should be taken into account when considering artificial infection protocols for A. galli.

A chromosome-level genome of the human blood fluke Schistosoma japonicum identifies the genomic basis of host-switching.

The evolution and adaptation of S.japonicum, a zoonotic parasite that causes human schistosomiasis, remain unclear because of the lack of whole-genome data. We construct a chromosome-level S.japonicum genome and analyze it together with 72 samples representing six populations of the entire endemic region. We observe a Taiwan zoophilic lineage splitting from zoonotic populations ∼45,000 years ago, consistent with the divergent history of their intermediate hosts. Interestingly, we detect a severe population bottleneck in S.japonicum, largely coinciding with human history in Asia during the last glacial maximum. We identify several genomic regions underlying natural selection, including GATAD2A and Lmln, both showing remarkable differentiation among different areas. RNAi knockdown suggests association of GATAD2A with parasite development and infection in definitive hosts, while Lmln relates to the specificity of the intermediate hosts. Our study provides insights into the evolution of S.japonicum and serves as a resource for further studies.

Local immune response of Canarian Majorera goats infected with Teladorsagia circumcincta

BackgroundDue to increased anthelmintic resistance, alternative methods to drugs are necessary to control gastrointestinal nematodes (GINs). Some of the most promising alternatives are based on the immune response of the host, such as the selection of genetically resistant breeds or the use of vaccines against these parasites. Given the limited information available on the immune response against GINs in goats, this study investigated the local immune response of goat kids of an indigenous Canary Islands breed (Majorera breed) experimentally infected with Teladorsagia circumcincta, one of the most pathogenic and prevalent GIN species.MethodsFor this purpose, the relationship between different parasitological (number of mature and immature worms, worm length, and number of intrauterine eggs) and immunological parameters at the local level (related to both the humoral and cellular immune response) was analyzed at early (1 week post-infection [wpi]) and late (8 wpi) stages of infection.ResultsPrimary infection of goat kids with T. circumcincta infective larvae (L3) generated a complex immune response that could be defined as Th2 type, characterized by increased infiltration in abomasal tissues of several effector cells as well as a progressive presence of specific antibodies against parasitic antigens in the gastric mucus. Cellular responses were evidenced from 1 wpi onward, showing an increase in antigen-presenting cells and various lymphocyte subsets in the gastric mucosa.ConclusionsThe complexity of the host response was evidenced by statistically significant changes in the number of all these subpopulations (MHCII+, CD4+, CD8+, γδ+, CD45R+, IgA+, and IgG+), as well as in the evolution of the relative cytokine gene expression. From a functional point of view, negative associations were observed between the number of most of the immune cells (CD4, IgA, IgG, and CD45R cells) and parameters that could be related to the fecundity of worms, a phenomenon that was especially evident when the number of IgG and CD45R cells or the specific IgA levels of the gastric mucus were compared with parasitological parameters such as the female worm length or fecal egg counts at 8 wpi.Graphical

New species of Guidus Ivanov, 2006 (Cestoda: Phyllobothriidea) from Bathyraja magellanica (Philippi) from the Patagonian Continental Shelf of Argentina.

Specimens representing two new species of Guidus Ivanov, 2006 were collected from the Magellan skate (Bathyraja magellanica [Philippi]) in the Patagonian Continental Shelf of Argentina, Southwestern Atlantic Ocean. Guidus francoi sp. n. and Guidus magellanicus sp. n. differ from their congeners by a particular combination of features, including type of bothridia, worm length, number of testes, and distribution of vitelline follicles. Guidus francoi sp. n. is distinguished from G. magellanicus sp. n. by having fewer proglottids, fewer testes and a higher ratio between the cirrus sac length and the proglottid width. The microthrix pattern of species of Guidus from the Southwestern Atlantic is described, based on specimens of G. francoi sp. n., Guidus magellanicus sp. n., and newly collected specimens of Guidus argentinense Ivanov, 2006. These three species share the presence of wide aristate gladiate spinitriches on the proximal bothridial surface, narrow gladiate spinitriches on the bothridial rim, and filitriches on the distal bothridial surface. The diagnosis of Guidus is revised to include several features exhibited by the new species (i.e., presence of bothridial indentations and bothridial stalks, distribution of vitelline follicles, and eggs grouped in cocoons). The discovery of G. francoi sp. n. and G. magellanicus sp. n. from B. magellanica increases the number of species of Guidus collected from batoids in the Southwestern Atlantic from one to three. The specificity exhibited by the species herein described reinforces the tight association between rays in the genera Guidus and Bathyraja.

Scanning electron microscopy as a confirmatory tool for morphological identification of Dirofilaria (Nochtiella) repens

Background: Dirofilaria repens (Railliet & Henry, 1911) is a filarial nematode that mainly affects canids. It can also infect humans accidentally, causing subcutaneous or conjunctival lesions, especially in the Old-World, including Egypt. Scanning electron microscopy (SEM) is very useful in diagnosis and identification of different parasites including D. repens. Objective: The current work aimed use SEM as a confirmatory tool for morphological identification of D. repens. Material and Methods: Morphological diagnosis of the excised nematodes was performed by studying their naked eye and light microscopic appearance. This was then confirmed using scanning electron microscopy (SEM). Results: Three worms were identified; one excised from maxillofacial subcutaneous lesion and two were excised from the bulbar conjunctival lesion. All worms were females. Scanning electron microscopic examination showed external cuticular longitudinal ridges and circular striations on the whole worm length surface especially in the mid-body region, which are characteristic for D. repens. The anterior end of the worms was wider than the posterior end that possessed a dimple-like anal orifice. The anterior end of the subconjunctival worms was spoon-like, and wider than that of the subcutaneous worm. The dimple-like anal orifice of the subconjunctival worm was more retracted than that of the subcutaneous worm. Conclusion: Two new human cases of dirofilariasis have been diagnosed in Egypt. SEM was used as a confirmatory tool for their diagnosis.

Zein-based nanocarriers for the oral delivery of insulin. In vivo evaluation in Caenorhabditis elegans.

The aim was to evaluate the potential of nanocarriers, based on the coating of zein nanoparticles (ZNP) with a Gantrez® AN-PEG conjugate (GP), for the oral delivery of insulin. ZNP-GP displayed less negative surface charge and a 14-fold higher diffusion coefficient in pig intestinal mucus than ZNP. Both nanoparticles showed a spherical shape and an insulin load of 77.5µg/mg. Under simulated gastric conditions, ZNP-GP released significantly lower amount of insulin than ZNP, while under simulated intestinal conditions, both types of nanoparticles displayed similar behaviour. In Caenorhabditis elegans wild-type N2, grown under high glucose conditions, insulin treatments reduced glucose and fat accumulation without altering the growth rate, the worm length, or the pumping rate. The effect was significantly greater (p<0.001) when insulin was nanoencapsulated in ZNP-GP compared with that encapsulated in ZNP or formulated in solution. This would be related to the highest capability of ZNP-GP to diffuse in the dense peritrophic-like layer covering intestinal cells in worms. In daf-2 mutants, the effect on fat and glucose reduction by insulin treatment was suppressed, indicating a DAF-2 dependent mechanism. In summary, ZNP-GP is a promising platform that may offer new opportunities for the oral delivery of insulin and other therapeutic proteins.

Enhancement in the ATP level and antioxidant capacity of Caenorhabditis elegans under continuous exposure to extremely low-frequency electromagnetic field for multiple generations

Purpose Safety concerns about the effects of long-term extremely low-frequency electromagnetic field (ELF-EMF) exposure on human health have been raised. To explore the effects of continuous exposure to ELF-EMF on organisms for multiple generations, we selected Caenorhabditis elegans as a model organism and conducted long-term continuous exposure studies for multiple generations under 20 °C, 50 Hz, and 3 mT ELF-EMF. Materials and methods Each generation of worms was treated with ELF-EMF from the egg in the same environment. After long-term exposure to ELF-EMF, the body length of the worms was detected, and 15th generation adult worms were selected as the research object. The ATP level and ATPase were detected, and the expression levels of genes encoding ATP synthase (r53.4, hpo-18, atp-5, unc-32, atp-3) were detected by RT-PCR. In worm’s antioxidant system, the level of reactive oxygen species (ROS) was detected by dichlorofluorescein staining, and the total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and catalase (CAT) activity were investigated. The expression of genes encoding superoxide dismutase (sod-1, sod-2, sod-3) was detected in adult (60 h) worms of the fifteenth generation (F15). Results These results showed that the body length of F15 worms increased significantly, ATP content increased significantly, ATP synthase activity was significantly enhanced, and the expression levels of the r53.4, hpo-18, atp-5, and atp-3 genes encoding ATPase were significantly upregulated in F15 worms. In addition, SOD activity increased significantly, and the expression levels of the sod-1, sod-2, and sod-3 genes encoding SOD were also significantly upregulated in F15 worms. Conclusions These results indicated that continuous exposure to 50 Hz, 3 mT ELF-EMF for multiple generations can increase the body length of worms, induce the synthesis of ATP and enhance the antioxidant capacity of worms.

Contracaecum osculatum (sensu lato) infection of Gadus morhua in the Baltic Sea: inter- and intraspecific interactions

The subpopulation of Atlantic cod, Gadus morhua, in the eastern part of the Baltic Sea has experienced a significant increase in infections with anisakid nematode larvae of the species Contracaecum osculatum sensu lato (s.l.) since the year 2000. The life cycle of the parasite includes seals and especially the grey seal, Halichoerus grypus, as final hosts, carrying the adult nematodes in the stomach, crustaceans (copepods, amphipods) as first intermediate hosts and various fish species (clupeids, sandeel) including cod as second intermediate/paratenic hosts. Cod with a body length below 28 cm are generally non-infected but experience increasing infection levels when they switch to a piscine diet (infected intermediate/paratenic hosts). We present an overall frequency distribution analysis of worms in 166 cod (body length 30–49 cm) collected in the spawning area over the last 5 years. It shows a fit to the negative binomial distribution, a prevalence of infection of 89.8%, a mean intensity of 29.3 parasites per fish (range 1–377) and a variance/mean ratio of 59.2 (≫1), indicating overdispersion. We present measurements of the adult Contracaecum osculatum (s.l.) specimens in the seal stomach and show that the parasites reach a maximum length of 6.6 cm (females) and 5.8 cm (males). L3s in sprat have a total length from 1to 11 mm whereas the larvae in cod liver are 3–27 mm. A decreasing mean worm length associated with high worm densities in cod (number of nematodes per liver) was recorded. Possible explanations might include timing of feeding on infected intermediate/paratenic hosts, intraspecific competition (crowding) between larvae in cod and host responses (indicated by a significant antibody production in cod against C. osculatum (s.l.) antigens). A significant negative correlation between infection intensity and muscle mass of cod was found, suggesting parasite-induced down-regulation of growth factors in cod.

Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity

A Trichinella spiralis aminopeptidase (TsAP) has been identified in intestinal infectious larvae (IIL) and adult worms (AW), but its biological function in the T. spiralis life cycle is unknown. The aim of this study was to characterize TsAP and ascertain its functions in the invasion, development and fecundity of T. spiralis. Recombinant TsAP (rTsAP) was expressed and purified. rTsAP has strong immunogenicity. qPCR and western blotting show that TsAP was transcribed and expressed at all T. spiralis lifecycle stages, but the expression level of TsAP mRNA and proteins at IIL and AW stages was obviously higher than those in muscle larvae (ML) and newborn larvae (NBL). The IFT results reveal that TsAP was principally located at the cuticle and the intrauterine embryos of this nematode. rTsAP had the enzymatic activity of natural aminopeptidase to hydrolyze the substrate Leu-pNA with an optimal temperature of 50 °C and optimal pH of 8.0. rTsAP promoted the larval penetration into intestinal epithelial cells, whereas anti-rTsAP antibodies suppressed the larval intrusion; the promotion and suppression was dose-dependently related to rTsAP or anti-rTsAP antibodies. TsAP protein expression level and enzymatic activity were reduced by 50.90 and 49.72% through silencing of the TsAP gene by specific siRNA 842. Intestinal AW and muscle larval burdens, worm length and female reproductive capacity were significantly declined in mice infected with siRNA-transfected ML compared to the control siRNA and PBS group. These results indicate that TsAP participates in the invasion, development and fecundity of T. spiralis and it might be a candidate target for anti-Trichinella vaccines.