Immature Worms Research Articles

Pathological Conditions Associated with Physaloptera Stomach Worm Infestation in Bengal Monitor Lizard Varanus bengalensis

The present study reports pathological conditions associated with Physaloptera sp. stomach worm infestation in Bengal monitor lizard (Varanus bengalensis). Four adult Bengal monitor lizards were rescued from the poachers and kept at Wild Animal Rescue and Rehabilitation Centre, Bannerghatta Biological Park, Bengaluru. Faecal samples from all were collected and examined. Physaloptera sp. ova were found in all faecal samples examined by sedimentation technique. One Bengal monitor lizard died during the period and systematically examined at necropsy for gross pathological lesions. Adult Physaloptera sp. nematodes were found to be firmly attached to the gastric mucosa causing erosive inflammatory foci. Histopathological examination of stomach revealed the presence of many immature and mature worms encapsulated within fibrous connective tissue. The intestinal villi were stunted and many cross sections of the worms could be demonstrated in the lumen. The worms also migrated to liver parenchyma and got encapsulated with fibrous connective tissue with infiltration of eosinophils in the sinusoids and the vascular tissue spaces. This seems to be the first report of Physaloptera sp. nematodiasis in Bengal monitor lizard kept under captivity for a period of 1.5 months.

Characterization of Lead Compounds Targeting the Selenoprotein Thioredoxin Glutathione Reductase for Treatment of Schistosomiasis.

Schistosomiasis is a widespread human parasitic disease currently affecting over 200 million people. Chemotherapy for schistosomiasis relies exclusively on praziquantel. Although significant advances have been made in recent years to reduce the incidence and intensity of schistosome infections, these gains will be at risk should drug-resistant parasites evolve. Thioredoxin glutathione reductase (TGR) is a selenoprotein of the parasite essential for the survival of schistosomes in the mammalian host. Several high-throughput screening campaigns have identified inhibitors of Schistosoma mansoni TGR. Follow up analyses of select active compounds form the basis of the present study. We identified eight compounds effective against ex vivo worms. Compounds 1-5 are active against all major species and development stages. The ability of these compounds to target immature worms is especially critical because praziquantel is poorly active against this stage. Compounds 1-5, 7, and 8 displayed schistosomicidal activity even after only 1 h incubation with the worms. Compounds 1-4 meet or exceed standards set by the World Health Organization for leads for schistosomiasis therapy activity. The mechanism of TGR inhibition was studied further with wild-type and mutant TGR proteins. Compounds 4-6 were found to induce an nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity in TGR, leading to the production of superoxide and hydrogen peroxide. Collectively, this effort has identified several active compound series that may serve as the basis for the development of new schistosomicidal compounds.

Human dirofilariasis: current situation and possibilities for diagnosis

Dirofilariasis in humans is a rarely diagnosed vector-borne zoonotic helminthic invasion. It is caused from filarial nematodes genus Dirofilaria & Railliet Henry, 1911 (Onchocercidae, Nematoda). Usually, the life cycle of the parasite takes place between mosquitoes (Culicidae) and carnivores. The human is an accidental host for the dirofilaria wherefore microfilariae in his blood circulation are almost always absent. The pathology of dirofilariasis results from the accidental localization of immature worms in the human body. Dirofilaria immitis caused heart and pulmonary dirofilariasis, but Dirofilaria repens, D. tenuis, and others can are found in different parts of the human body. The spread and Dirofilaria-invasive rates аrе undergone significant modifications affected by global climate change. Surgical extraction of the parasite in humans usually has a therapeutic effect, after which an etiological diagnosis is possible. The aim of the study is to review the current situation of human dirofilariasis in clinical and epidemiological aspects and possibilities for diagnosis.

Feline asthma and heartworm disease: Clinical features, diagnostics and therapeutics.

For feline practitioners, the cat with a cough or respiratory distress and thoracic radiographs with a bronchial or bronchointerstitial pattern suggests lower airway disease. Two important differentials, allergic asthma and heartworm disease (HWD), have many overlapping clinicopathologic features, but also clear and important differences in terms of cause and disease progression, treatment and prognosis. Notably, asthma is readily treatable and HWD is preventable. Feline HWD comprises two clinical syndromes: the comparatively recently described heartworm-associated respiratory disease (HARD) and adult HWD. The former is much more common; very few cats with HARD develop adult HWD. In HARD, following death of immature worms, pulmonary lesions may improve over time ('self-cure'). Lesions of adult HWD also improve over time as long as reinfection does not occur; however, with death of adult heartworms, mortality is high, and the prognosis is guarded. In asthma, morbidity is relatively high, but mortality is low, with an overall good to excellent prognosis. Feline asthma is encountered worldwide. In the authors' impression, feline HWD is often under-recognized. The aim of this review is to assist clinicians in differentiating feline asthma from feline HWD; as such, the emphasis is on distinguishing clinical features, as well as on diagnostics, therapy and prognosis. In differentiating these conditions, clinicians can attempt the goal of properly managing these diseases and can best educate owners on prognosis. For both feline asthma and feline HWD, the authors have drawn on the available peer-reviewed literature studies involving experimental models as well as spontaneous disease.

Acanthocephalans from Marine Fishes from Patagonia, Argentina

In this study, 542 individual fish from 20 species from the Patagonian continental shelf of Argentina were examined for acanthocephalans. A total of 1,547 acanthocephalans belonging to 5 species were collected from 18 species of fish. Adult forms were represented by 2 species: Aspersentis johni ( Baylis, 1929 ) (Heteracanthocephalidae) from longtail southern cod, Patagonotothen ramsayi (Regan) (new host record), and Breizacanthus aznari Hernández-Orts, Alama-Bermejo, Crespo, García, Raga and Montero, 2012 (Arhythmacanthidae) from raneya, Raneya brasiliensis (Kaup). Immature worms of B. aznari were also collected from the intestine of pink cusk-eel, Genypterus blacodes (Forster) (new host record). Cystacanths of 3 species of Corynosoma Lühe, 1904 (Polymorphidae) were found encapsulated in the mesenteries of fish. Corynosoma australe Johnston, 1937 was the most abundant acanthocephalan in our study, infecting 18 species of fish and accounting for >89.9% of all specimens collected. A cystacanth of Corynosoma bullosum (Linstow, 1892) was found in "castañeta", Nemadactylus bergi (Norman) (new host record), and cystacanths of Corynosoma cetaceum Johnston and Best, 1942 were collected from red searobin, Prionotus nudigula Ginsburg, and flounders Paralichthys isosceles Jordan (new host record) and Xystreurys rasile (Jordan). The Patagonian shelf of Argentina represents a new locality record for A. johni and C. bullosum. This survey is a starting point for understanding the diversity of marine acanthocephalans in Patagonian waters.

In silico repositioning of approved drugs against Schistosoma mansoni energy metabolism targets

Schistosomiasis is a neglected parasitosis caused by Schistosoma spp. Praziquantel is used for the chemoprophylaxis and treatment of this disease. Although this monotherapy is effective, the risk of resistance and its low efficiency against immature worms compromises its effectiveness. Therefore, it is necessary to develop new schistosomicide drugs. However, the development of new drugs is a long and expensive process. The repositioning of approved drugs has been proposed as a quick, cheap, and effective alternative to solve this problem. This study employs chemogenomic analysis with use of bioinformatics tools to search, identify, and analyze data on approved drugs with the potential to inhibit Schistosoma mansoni energy metabolism enzymes. The TDR Targets Database, Gene DB, Protein, DrugBank, Therapeutic Targets Database (TTD), Promiscuous, and PubMed databases were used. Fifty-nine target proteins were identified, of which 18 had one or more approved drugs. The results identified 20 potential drugs for schistosomiasis treatment; all approved for use in humans.

The benign helminth Hymenolepis diminuta ameliorates chemically induced colitis in a rat model system.

The tapeworm Hymenolepis diminuta is a model for the impact of helminth colonization on the mammalian immune system and a candidate therapeutic agent for immune mediated inflammatory diseases (IMIDs). In mice, H. diminuta protects against models of inflammatory colitis by inducing a strong type 2 immune response that is activated to expel the immature worm. Rats are the definitive host of H. diminuta, and are colonized stably and over long time periods without harming the host. Rats mount a mild type 2 immune response to H. diminuta colonization, but this response does not generally ameliorate colitis. Here we investigate the ability of different life cycle stages of H. diminuta to protect rats against a model of colitis induced through application of the haptenizing agent dinitrobenzene sulphonic acid (DNBS) directly to the colon, and monitor rat clinical health, systemic inflammation measured by TNFα and IL-1β, and the gut microbiota. We show that immature H. diminuta induces a type 2 response as measured by increased IL-4, IL-13 and IL-10 expression, but does not protect against colitis. In contrast, rats colonized with mature H. diminuta and challenged with severe colitis (two applications of DNBS) have lower inflammation and less severe clinical symptoms. This effect is not related the initial type 2 immune response. The gut microbiota is disrupted during colitis and does not appear to play an overt role in H. diminuta-mediated protection.

Using natural clinoptilolite zeolite as an amendment in vermicomposting of food waste.

The effect of adding different proportions of natural clinoptilolite zeolite (5 and 10%) to food waste vermicomposting was investigated by assessing the physicochemical characteristics, worms' growth, and maturation time of finished vermicompost in comparison with the vermicompost prepared with no amendment (control). Vermicomposting was performed in 18 plastic containers for 70days. The experimental results showed that the carbon-to-nitrogen (C/N) ratios were 15.85, 10.75, and 8.94 for 5 and 10% zeolite concentration and control after 70days, respectively. The addition of zeolite could facilitate organic matter degradation and increase the total nitrogen content by adsorption of ammonium ions. Increasing the proportion of zeolite from 0% (control) to 10% decreased the ammonia escape by 25% in the final vermicompost. The natural zeolite significantly reduced the electrical conductivity (EC). At the end of the process, salinity uptake efficiency was 39.23% for 5% zeolite treatment and 45.23% for 10% zeolite treatment. The pH values at 5 and 10% zeolite-amended treatments were 7.31 and 7.57, respectively, in comparison to 7.10 in the control. The maturation time at the end of vermicomposting decreased with increasing zeolite concentration. The vermicompost containing 5 and 10% zeolite matured in 49 and 42days, respectively, in comparison to 56days for the control. With the use of an initial ten immature Eisenia fetida worms, the number of mature worms in the 10% zeolite treatment was 26 more than that in the 5% zeolite treatment (21 worms) and 9 more than that in the control treatment (17 worms). Significantly, natural zeolite showed a beneficial effect on the characteristics of the end-product when used in the vermicomposting of food waste.

Phthalimido-thiazole as privileged scaffold: activity against immature and adult worms of Schistosoma mansoni.

Phthalimide, 1,3-thiazole, and thiazolidinone cores are considered privileged scaffolds and represent an attractive starting point to design new bioactive compounds for neglected tropical disease (NTD). Schistosomiasis is a NTD, caused by Schistosoma worms which praziquantel (PZQ) is the only drug used to treat humans, but the decrease in the effect after treatment has been reported. Recently, some phthalimide-thiazole derivatives exhibited in vitro antischistosomal activity against adult worms with significant ultrastructural changes and a lower cytotoxic effect on splenocytes. This new biological phthalimido-thiazole profile has motivated us to evaluate a new generation of such molecules in immature and adult worms. Thus, a phthalimido-thiazolidinone derivative, (3c), and three phthalimido-thiazoles (6c, 7a, and 7h) were evaluated concerning their in vitro activity on schistosomulae and adult worms. The results showed that these compounds brought a significant reduction on the mortality, inhibited oviposition, and then induced mortality in immature and adult worms alike. According to scanning electron microscopy, the tegument was the principal target for 7a and 7h and revealed gradual damage to the tegument surface, inducing destruction and decomposition of the tegument in the same areas and exposition of subtegumental tissue and of muscle tissue. Furthermore, they caused less toxicity in splenocytes than PZQ. Compounds 7a and 7h revealed to possess promising activity against larval forms. According to the present study, the privileged structure phthalimido-thiazoles act as a molecular framework that has antischistosomal activity and most form the basis to the next pre-clinical tests. Graphical abstract.

Evaluation of nanoselenium and nanogold activities against murine intestinal schistosomiasis

Nanomedicine is one of the most important methods used to treat human diseases including parasitic diseases. Schistosomiasis is a major parasitic disease that affects human health in tropical regions. Whilst Praziquantel is the main classic antischistosomal drug, new drugs are required due to the poor effect of the drug on the parasite juveniles and immature worms, and the emergence of drug resistant strains of Schistosoma. The present study aimed to examine the curative roles of both gold and selenium nanoparticles on jejunal tissues of mice infected with Schistosoma mansoni. Transmission electron microscopy was used for characterization of nanoparticles. Gold nanoparticles of 1 mg/kg mice body weight and selenium nanoparticles 0.5 mg/kg body weight were inoculated separately into mice infected with S. mansoni. The parasite induced a significant decrease in glutathione levels; however, the levels of nitric oxide and malondialdehyde were significantly increased. Additionally, the parasite introduced deteriorations in histological architecture of the jejunal tissue. Treatment of mice with metal nanoparticles reduced the levels of body weight changes, oxidative stress and histological impairment in the jejunal tissue significantly. Therefore, our results revealed the protective role of both selenium and gold nanoparticles against jejunal injury in mice infected with S. mansoni.

Effect of Nigella sativa oil on Schistosoma mansoni immature worms in experimentally infected mice

Background Nigella sativa oil is one of the promising drugs of a plant origin that have an antischistosomal effect. Aim The aim of the present work was to explore the role of N. sativa oil either alone or in combination with praziquantel on immature worms of Schistosoma mansoni. Materials and methods N. sativa oil capsules dissolved in corn oil were administered to S. mansoni-infected mice aiming to study the therapeutic and prophylactic effects. The study included six groups of mice: (a) noninfected and nontreated, (b) infected and nontreated, (c) prophylactic N. sativa oil group, (d) praziquantel group, (e) therapeutic N. sativa oil group, and (d) combined group. All mice were killed 4 weeks after infection and treatment. The antischistosomal effect of N. sativa oil was assessed by worm burden, histopathology, and scanning electron microscopy. Results Therapeutic N. sativa oil group showed the highest significant decrease in the mean number of immature worm burden (R% was 57%) when given after infection followed by the combined group (R%=55.67%), whereas prophylactic N. sativa oil group showed nonsignificant decrease (R%=37%). The least changes were observed in praziquantel group (R%=0%). Conclusion N. sativa oil has a potent effect on immature S. mansoni which could be helpful for potentiating praziquantel effect and thus reducing development of resistance.

Californian red worm biomass increase and its cobalt accumulation under different concentrations of the metal in nutrient medium

<p>Californian red worms hybrid biomass is a biologically valuable feed supplement for livestock, poultry and fish. The worm body contains a significant amount of protein, lipids, vitamins and minerals. The chemical composition of worms depends on the nutrient medium in which they were grown. Worms can accumulate minerals from the nutrient medium in the body which called forth conducting research on obtaining worms biomass enriched with Cobalt with the prospect of its further use in fish feeding.<br />It has been found out that the worms quantity and the mass increase depends on the content of Cobalt in the nutrient medium. Adding 20 mg / kg of Cobalt to the nutrient medium contributed to 38.0% and 40.4% increased amount of worms weighing 0.4-0.8 g respectively. The study reveals that the number of immature worms increased by 32.2%. Adding 40 mg / kg of Cobalt to the nutrient medium resulted in worms number and mass increased by 45.9 and 51.1% respectively. Adding 160 mg / kg of Cobalt resulted in 6.5-27.7% smaller amount of worms weighing 0.4-0.8 g as compared with the experimental groups. It has been found out that the number of small worms reduced by 24.0-50.7% compared with the experimental groups under adding 160 mg / kg of Cobalt. The weight reduced by 22.4% and amount of young worms compared with the control has been revealed.<br />We have established the pattern that as Cobalt amount in the nutrient medium increases, its concentration in the worms biomass increases as well. The highest content of Cobalt was found in the biomass dry matter of worms grown in the nutrient medium enriched with the studied metal in the amount of 160 mg / kg.<br />Biomass of worms grown in the nutrient medium with 40 mg / kg of the studied metal added can be used in fish feeding as a protein supplement with a high content of Cobalt.</p>

Epidemiological investigation of a severe rumen fluke outbreak on an Irish dairy farm.

Although the rumen fluke, Calicophoron daubneyi is now very common and widespread throughout Western Europe, reports of clinical cases are still rare. This study explores the epidemiological background to a severe rumen fluke outbreak in 6-month-old heifers on a dairy farm in Ireland. Sequence analysis of the cytochrome oxidase subunit 1 (Cox1) gene of the rumen fluke metacercariae on pasture failed to identify predominant, possibly pathogenic subtypes. However, estimates of metacercarial load indicated that the animals were exposed to a daily dose of about 5334 C. daubneyi metacercariae for a period of 3 weeks resulting in the build-up of very large numbers of immature worms in the small intestine. It is hypothesized that specific environmental conditions may favour this parasite over its competitor, the liver fluke, Fasciola hepatica, possibly by allowing it to emerge earlier. The possibility that C. daubneyi may be better adapted to the Irish climate than F. hepatica together with the fact that selective treatment against F. hepatica effectively frees the niche for C. daubneyi, may result in the gradual replacement of F. hepatica by C. daubneyi.

The progression of heartworm associated respiratory disease (HARD) in SPF cats 18\xa0months after Dirofilaria immitis infection

BackgroundHeartworm-associated respiratory disease (HARD) in cats is induced by the arrival and death of immature adult Dirofilaria immitis in the pulmonary system and is indistinguishable from mature adult heartworm infection.MethodsA controlled, blind research study investigated the long-term (18 months post infection, PI) consequences of the inflammatory response associated with the death of immature adult heartworms in cats.Three groups of cats, 10 per group, were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated with selamectin (Revolution®; Zoetis) per label directions at 28 days PI and once monthly for 17 months. Group B cats were treated orally with ivermectin (Ivomec®; Merial) at 150 μg/kg) at 70 days PI, then every 2 weeks for 15 months. Group C cats were untreated PI. At baseline (Day 0) and on Days 70, 110, 168, 240, 309, 380, and 505 PI, peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected.ResultsThe selamectin-treated cats (Group A) and ivermectin-treated cats (Group B) were free of heartworms or heartworm fragments at necropsy. All cats became heartworm antibody positive at some time point in the study except for one cat in Group A. Only cats in Group C (all with adult heartworms) were heartworm antigen positive. The heartworm antibody titer for Group B was highest on Days 110 to 168 and then decreased over time and 50% were serologically antibody negative on Day 240. Eosinophilic bronchoalveolar lavage (BAL) cytology and peripheral eosinophilia were most pronounced on Day 110 in all cats. Randomly distributed myofibrocytes in the lungs of some Group A cats suggest that precardiac larval stages were affecting the lungs. Radiographs in Group B cats demonstrated partial resolution of the initial HARD reaction but chronic myofibrocyte proliferation was histologically evident 18 months after infection.ConclusionHARD was induced by immature adult worm infection with progressive improvement starting 6 to 8 months after infection but histologic lesions were evident in some cats 18 months after infection. The serologic antibody assay was negative in 50% of cats at 8 months and 100% of cats at 18 months post infection. Abnormal radiographic lung patterns continued in a subset of Group B cats for months after heartworm antibody serology and BAL cytology returned to normal.

Heartworm-associated respiratory disease (HARD) induced by immature adult Dirofilaria immitis in cats

BackgroundA controlled, blind research study was conducted to define the initial inflammatory response and lung damage associated with the death of immature adult Dirofilaria immitis in cats as compared with cats developing adult heartworm infections and cats on preventive medication.MethodsThree groups of cats were utilized, 10 per group. All cats were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated topically with selamectin (Revolution®; Zoetis) per label directions at 28 days post infection (PI) and once monthly for 8 months. Group B cats were treated orally with ivermectin (Ivomec®; Merial) at 150 μg/kg at 70 days PI, then every 2 weeks for 5 months. Group C cats were untreated PI. At baseline (Day 0) and on Days 70, 110, 168, and 240 PI, peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected on all cats. Upon completion of the study (Day 245), cats were euthanized and necropsies were conducted.ResultsResults were analyzed statistically between groups by ANOVA and by paired sample T testing for changes within the group over time. The selamectin-treated cats (Group A) did not develop radiographically evident changes throughout the study and were free of adult heartworms or worm fragments at necropsy. The heartworm life cycle was abbreviated with oral doses of ivermectin (Group B), shown by the absence of adult heartworms or worm fragments at necropsy. The early stage of immature adult worm in Group B cats, however, did induce severe pulmonary airway, interstitial, and arterial lung lesions, revealing that the abbreviated infection is a significant cause of respiratory pathology in cats. Cats in Groups B and C could not be differentiated based on radiographic changes, serologic antibody titers, complete blood count, or bronchoalveolar lavage cytology at any time point throughout the study. Eighty percent of cats in Group A and 100% of cats in Groups B and C became heartworm antibody positive at some time point post infection.ConclusionsThe clinical implications of this study are that cats that become infected with immature adult heartworms may not develop fully mature heartworms and are only transiently heartworm antibody positive, but do develop Heartworm-Associated Respiratory Disease (HARD).

Species composition of larvae cultured after anthelmintic treatment indicates reduced moxidectin susceptibility of immature Cylicocyclus species in horses

For the control of cyathostomins in horses, the macrocyclic lactones (MLs), moxidectin (MOX) and ivermectin (IVM) are the most commonly used anthelmintics. However, reduced activity, observed as shortening of the egg reappearance period (ERP) has been described. Shortening of the ERP may be caused by a decreased susceptibility of immature worms for MLs. Alternatively, immature worms may develop faster into egg producing adults as a result of repeated ML treatments. The species composition of the larval cultures obtained shortly after ML and pyrantel (PYR) treatment can confirm the hypothesis of decreased ML susceptibility, as this is often class-specific, whereas faster development would also occur after treatment with anthelmintics with a different mode of action. From 3 farms with a known history of shortened ERP, 8 horses per farm were selected and divided into 2 groups. The MOX-PYR-MOX group was treated twice with MOX (day 0 and 126) and once with PYR (day 84) and the IVM-PYR-IVM group was treated twice with IVM (day 0 and 98) and once with PYR (day 56). Cultured infective larvae (L3s) were counted and differentiated with the reverse line blot on pooled samples. Per cyathostomin species, the number of larvae per gram was calculated. The efficacy of all ML treatments was 100% and a shortened ERP was found on all 3 farms. The species composition of the larval cultures after ML treatment did not differ significantly from that after PYR treatment in the IVM-PYR-IVM group, but it did differ in the MOX-PYR-MOX group. The larval cultures obtained after MOX treatment consisted mostly of Cylicocyclus nassatus, while after PYR treatment Cylicostephanus longibursatus was the most abundant species. In the cultures from 42days after MOX treatment 6 cyathostomin species from 3 genera were found on the farm with the lowest activity (farm 1), while on the farm with the highest activity (farm 3) only 3 species from one genus were found in the same number of examined L3s. The high numbers of L3s of Cylicocyclus species 42days after MOX treatment and the low numbers 42days after PYR treatment can be explained by reduced susceptibility of the immature worms to MOX, but not by a faster development. In conclusion, shortening of the ERP following MOX treatment is most likely a process in which an increasing number of immature worms from an increasing number of species is becoming less susceptible to the active compound.

Review of the genus Shirleyrhynchus Beveridge & Campbell, 1988 (Trypanorhyncha: Shirleyrhynchidae), with the resurrection of S. butlerae Beveridge & Campbell, 1988 and the description of S. panamensis n. sp.

A new species of trypanorhynch cestode is described from two species of stingrays, the Panamic stingray Urotrygon aspidura (Jordan & Gilbert) and the Pacific chupare Himantura pacifica (Beebe & Tee-Van) collected in the Golfo de Montijo in the Eastern Pacific Ocean off the coast of Panama. Shirleyrhynchus panamensis n. sp. represents an important addition to the family, which until now consisted of two monotypic genera. The new species is characterised by an elongate scolex with four, ovate bothria, presence of prebulbar organs, absence of gland cells within the muscular bulbs and an oncotaxy with a typical heteroacanthous, heteromorphous tentacular armature, a characteristic basal armature and the presence of a slight basal swelling. It is readily distinguished from its congeners by a smaller scolex and features of the oncotaxy, such as dissimilar hooks on opposing principle rows, a commencement of hook rows from the bothrial to the antibothrial surface and a much shorter basal armature. Although described only on the basis of immature worms lacking a strobila, the new species adds information on features of the oncotaxy within Shirleyrhynchus Beveridge & Campbell, 1988. Observation of the holotype of Shirleyrhynchus aetobatidis (Shipley & Hornell, 1906) revealed apparent differences from Australian specimens that have been described as Shirleyrhynchus butlerae Beveridge & Campbell, 1988 but which were later synonymised. Observations of type-specimens of S. butlerae also revealed differences from the original description and some morphological characteristics are amended. Shirleyrhynchus butlerae is herein resurrected and an amended generic diagnosis and a key to the identification of species are provided. The molecular voucher specimen of 'S. aetobatidis' utilised in previous molecular phylogenetic studies was re-observed which revealed a misidentification of the specimen with Parachristianella indonesiensis Palm, 2004.

PROPOLIS AS A NATURAL FEED ADDITIVE IN RUMINANT DIETS; CAN PROPOLIS AFFECT THE RUMINANTS PERFORMANCE?: A REVIEW

Propolis is a resinous substance collected by honeybees from buds and leaves of trees and plants, mixing with pollen as well as enzymes secreted by bees. Propolis is known to be active against gram positive bacteria, viruses, fungi, oxidants, inflammation, tumors, and parasites thus recently it have been considered among the natural feed alternative to antibiotics in ruminant diets. Propolis extracts were found to have a promising effect on decreasing energy lost as methane and total N losses, the most significant inefficiencies remaining in ruminant production systems that may limit production performance and contribute to the release of pollutants to the environment. However, these effects are divers among propolis types. In vitro, however alamo and green propolis were extracted with the same method, but alamo propolis presented a higher efficiency to reduce methane production than the green one and both of them had no effect on ammonia concentrations, gas production and ruminal degradability. Red and brown propolis extracted similarly had very different compositions and seemed to act in a similar way in modifying the ruminal degradation of nutrients and mitigating methane formation. In vivo studies involving ruminant metabolism, divers' effects of different propolis samples have been found on the total digestibility, ruminal digestibility, intestinal digestibility. However, similar effects were observed with the different propolis studies on reducing ammonia production and the number of ruminal ciliate protozoa in buffaloes and dairy cows, as well as in in viro trails. The inhibitory activity of different propolis samples were observed on both mature and immature hepatic worms, and intestinal eggs. Red propolis had good impact on general ewe health during critical periods such as flushing and found to have high efficacy against the gastro intestinal helminthiasis. Similar findings in growth performance in a study on feedlot lambs fed a diet supplemented with brown propolis, but the green propolis was not efficient. Often, in most studies, the type or the chemical composition of the propolis is not even specified in publications. The biological activity of propolis may vary, together with its chemical composition, as it comprises numerous constituents together with the method of extraction and concentrations of the solvent and propolis. Thus in order to ensure the propolis quality as feed additive and to be officially accepted by major health agencies, propolis requires a chemical standardization and identify the active component(s) of propolis and subsequently analyze these in commercial batches.

Dirofilaria immitisin a child from the Russian Federation

An immature female worm, Dirofilaria immitis, was isolated from the scrotum of a 14-month-old child. This is the first identification of human dirofilariosis caused by D. immitis in a relatively Northern region (Moscow) of the Russian Federation. The parasite was diagnosed by means of morphological examination of the worm, confirmed by PCR assay. This case raises questions about the range of the D. immitis distribution among humans in Russia. To better understand the geographical distribution of dirofilarioses, detailed clinical and epidemiological information should be collected from human cases with appropriate laboratory confirmation.

The morphology of free-living stages and immature parasites of Rhabdias paraensis (Nematoda: Rhabdiasidae), a parasite of Rhinella marina (Anura: Bufonidae) in Brazil.

Rhabdias paraensis Santos, Melo, Nascimento, Nascimento, Giese et Furtado, 2011 was described based on fully gravid worms. Further investigations on the free-living stages, immature worms and young individuals were facilitated by cultivation in the laboratory, which allowed us to add new information about the morphology and development of the species. Observations on the free-living development of R. paraensis showed that the life cycle is typical of Rhabdias, with alternation of gonochoristic and hermaphroditic generations and without homogony. Males of the free-living generation were different from those in several species of the genus studied previously. In the original description, the excretory glands and duct were absent in gravid specimens of R. paraensis, while in this study, distinct excretory glands and a duct were observed in immature and young individuals. Additionally, we recognised the separation of the buccal capsule walls into anterior and posterior portions and described the specific shapes of these portions in lateral and apical view. Studies on the morphology and development of free-living stages of Rhabdias spp. from Neotropical regions may provide additional information for species determination.