Occurrence of nematodes and anthelmintic management of ostrich farms from different Brazilian states: Libyostrongylus douglassii dominates mixed infections

Ostriches were imported to Brazil in the early 1990s without appropriate parasite control leading to a co-importation of parasites. Libyostrongylus douglassii, Libyostrongylus dentatus and Codiostomum struthionis are nematodes of the ostriches that have been reported on some Brazilian farms but no national survey has been performed. Moreover, little is known about anthelmintic usage to control these parasites in Brazil. Therefore, the occurrence of L. douglassii, L. dentatus and C. struthionis and the anthelmintic management practices in Brazilian ostrich farms were studied. A questionnaire about management practices and information on how to collect, store and ship feces to the laboratory were sent to 17 farms located in 9 Brazilian states. Received fecal samples were submitted to fecal cultivation and larval harvested. Infective larvae were morphologically analyzed and quantified. The majority of the farms (88%) used an anthelmintic at least once a year. Ivermectin was the principal anthelmintic. Libyostrongylus sp. and C. struthionis were found in samples from 76 and 18% of the examined farms, respectively. The presence of L. douglassii (61-97%) was higher than L. dentatus (3-57%) and C. struthionis (2-30%). At the farm level, both species of Libyostrongylus were always found together. Sheath tail measurements of their infective larvae were crucial for morphological diagnosis. L. dentatus and L. douglassii are well adapted to Brazil and are widely distributed in the Brazilian territory, proving that this mixed infection in Brazilian ostrich's farms is common.     

The occurrence of Codiostomum struthionis in ostriches (Struthio camelus) of different ages and during the dry and rainy seasons at two farms in the State of Rio de Janeiro, Brazil

Ostriches are susceptible to a wide variety of diseases that can affect their commercial breeding. The aim of this study was to identify morphologically Codiostomum struthionis and examine its seasonal occurrence among animals of three age categories in two commercial farms in Areal and Itaboraí, State of Rio de Janeiro, Brazil. Fresh fecal samples were collected monthly, from June 2004 to May 2006. Eggs per gram of feces (EPG) were counted after centrifugal flotation in saturated sugar solutions and infective larvae were evaluated after coproculture. A total of 432 fecal samples were collected from three groups according to ostrich age: up to 90 days, 91-365 days and over 365 days (adults). The dry and rainy seasons were defined according to official meteorological data. Typical eggs of the Strongylida order were identified in 242 samples (235 from adult ostriches). Once obtained, infective larvae were identified as C. struthionis. Adult ostriches exhibited significantly greater numbers of eggs in their feces during the rainy season compared to the dry season (p<0.05). In the Areal farm, significant (p<0.05) values were found when adults ostriches EPG were compared to ostriches up to 90 days and also for ostriches from 91 to 365 days. When comparing adult EPGs between the two commercial breeding farms, Areal exhibited greater EPGs during both the dry (p=0.0215) and rainy (p=0.0011) seasons.     

Morphological diagnosis of infective larvae of Libyostrongylus douglassii (Cobbold, 1882) Lane, 1923 and L. dentatus Hoberg, Lloyd and Omar, 1995 (Nematoda: Trichostrongylidae) of ostriches

The differentiation of the species of the Libyostrongylus genus is only possible with the obtainment of the adult parasites in the ostriches proventriculus and gizzard. The present work confirms that it is possible to differentiate the infective larvae of L. douglassii and L. dentatus allowing the differential diagnosis of these species by fecal culture. To show this, adult females from both species were collected from ten proventriculus from adult ostriches and separated by species. Both groups were macerated individually added to sterilized feces for standard fecal cultures. The infective larvae were recovered, identified, quantified and measured. All proventriculus analyzed were parasitized by Libyostrongylus spp. and a clear heterogeneous location for each species was observed. The infective larvae from the fecal cultures of macerated L. douglassii presented a mean total length of 874.3+/-33.80 microm, and a short sheath tail (29.5+/-4.11 microm) with acute termination. The infective larvae from the macerated L. dentatus presented mean total length of 856.0+/-43.63 microm, long sheath tail (61.2+/-9.52 microm) with filamentous termination. The mean measures of the tails of both species had a significant difference. The differentiation of the infective larvae of L. douglassii and L. dentatus by fecal cultures will facilitate the diagnosis of both species for further understanding the Libyostrongylus biology.     

Occurrence of Libyostrongylus sp. (Nematoda) in ostriches (Struthio camelus Linnaeus, 1758) from the north region of the state of Rio de Janeiro, Brazil

Domestic production of ostrich in Brazil started in the beginning of the last decade, but its sanitary state has not been reported. Libyostrongylus sp. is an ostrich specific nematode whose parasitism can severely affect the birds. Thus, Libyostrongylus spp. larvae were identified in commercial ostriches in the north region of the state of Rio de Janeiro, Brazil. The EPG was determined and fecal cultivation was performed. The eggs presented typical characteristics of strongylid and were present in five out of six farms. The mean EPG varied from 22 to 2395 and Libyostrongylus spp. prevalence was from 0 to 100%, with adult birds more infected. Two types of infective larvae with tail finishing in a tipped spiny knob were distinguished. The first had a mean length of 848 microm (710-1010) with a long sheath tail of about 66 microm (52-112). The other had a mean length of 826 microm (620-940) with a short, more abruptly ending sheath tail of 32 microm (22-40) and a less rounded cephalic end. The differences between these larvae suggest two Libyostrongylus species.     

Winter survival in Sweden of L(3)-stage larvae of the ostrich wireworm Libyostrongylus douglassii

Libyostrongylus douglassii, a pathogenic trichostrongylid nematode of the ostrich, was recently identified as a potentially important parasite in ostriches on Swedish farms. This parasite is well adapted to the hot and dry climates of sub-Saharan Africa, the natural habitat of the ostrich. The influence of low temperatures in colder climates, such as in Sweden, on free-living stages of L. douglassii is however insufficiently known. In this study, parasite free ostrich faeces were mixed with infective L(3)-stage larvae of L. douglassii, which had been cultured from eggs obtained from two Swedish farms. Samples of the mixture were placed on a grass surface, and analyses for L(3)-stage larvae were performed at regular intervals. The results of the study showed that L(3)-stage larvae may remain alive during a period of at least 97 days on pasture during the winter season in Sweden, even when the temperature within the samples was repeatedly below -0.1 degrees C, and the air temperature varied between -19.6 and +14.9 degrees C. It was concluded that L(3)-stage larvae of L. douglassii probably have the ability to remain viable on pastures during winter in Sweden.     

Inactivation of eggs and larvae of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora after passage in pigs

The study investigated the effect of gastrointestinal passage in pigs on free-living stages of bovine nematodes. Two Landrace x Yorkshire pigs, A and B, were fed fresh eggs of Ostertagia ostertagi and Cooperia oncophora while two other pigs, C and D, were fed third stage larvae (L3) of the same parasites. Faeces from the pigs were collected for 48 h after ingestion. In pigs A and B, 15 and 66% of the eggs were recovered after passage, respectively. However, only 0.003 and 0.002% of the ingested eggs developed into third stage larvae (L3) after subsequent culturing. In pigs C and D, 0.01 and 0.02% of the L3 survived the passage of the gastrointestinal tract. Fresh O. ostertagi and C. oncophora eggs were cultured in parasite free porcine and bovine faeces. Only 0.05% L3 developed in porcine faeces, whereas 21% of the eggs developed into L3 in the bovine culture. Our results demonstrate an extremely poor rate of development and survival of both bovine nematode eggs and infective larvae after passage in pigs. It may imply that pigs can play an important role in reducing transmission of cattle nematodes if the two species are grazed together or alternately.     

Effects of supplemental feeding on gastrointestinal parasite infection in elk (Cervus elaphus): preliminary observations

The effects of management practices on the spread and impact of parasites and infectious diseases in wildlife and domestic animals are of increasing concern worldwide, particularly in cases where management of wild species can influence disease spill-over into domestic animals. In the Greater Yellowstone Ecosystem, USA, winter supplemental feeding of Rocky Mountain elk (Cervus elaphus) may enhance parasite and disease transmission by aggregating elk on feedgrounds. In this study, we tested the effect of supplemental feeding on gastrointestinal parasite infection in elk by comparing fecal egg/oocyst counts of fed and unfed elk. We collected fecal samples from fed and unfed elk at feedground and control sites from January to April 2006, and screened all samples for parasites. Six different parasite types were identified, and 48.7% of samples were infected with at least one parasite. Gastrointestinal (GI) nematodes (Nematoda: Strongylida), Trichuris spp., and coccidia were the most common parasites observed. For all three of these parasites, fecal egg/oocyst counts increased from January to April. Supplementally fed elk had significantly higher GI nematode egg counts than unfed elk in January and February, but significantly lower counts in April. These patterns suggest that supplemental feeding may both increase exposure and decrease susceptibility of elk to GI nematodes, resulting in differences in temporal patterns of egg shedding between fed and unfed elk.     

Habronemiasis: Biology,Signs, and Diagnosis,and Treatment and Prevention of the Nematodes and Vector Flies

Habronemiasis is a parasitic disease of horses and other equids which is caused by the invasion of Draschia megastoma, Habronema majus, and H. muscae nematodes and is transmitted by house flies, face flies, and stable flies. The adult nematodes live in the horse's stomach, laying eggs, which pass out in the horse's feces. The carrier flies lay eggs in the horse's manure, which becomes infested with larvae while the flies feed in the manure. The nematode larvae molt while in the fly larvae and develop into infective L3 larvae by the time the adult flies emerge from their pupal stage. The infective larvae are then transmitted to the horse when flies feed on secretions or discharges from hosts' eyes, nostrils, mouths, wounds, and other openings. The L3 larvae are ingested by the horse and are then able to complete their lifecycle. The most common aberrant forms (conjunctival and cutaneous habronemiasis) are associated with the nematode larvae being deposited in these areas, not completing its life cycle, with resultant signs probably associated with local hypersensitivity. This article details the biology of nematode parasites, diagnosis and treatment of this condition, and methods of prevention and control of the transmitting flies.     

Parasites from farmed ostriches (Struthio camelus) and rheas (Rhea americana) in Europe

During a 4-year-period, more than 500 ostriches and several rheas, all born in European countries and raised in Spain and Portugal, have been analyzed for the presence of ectoparasites and endoparasites. A total of 29 parasite species have been found, most of them of the gastrointestinal tract. Some of the helminth species found may represent spureous parasitosis, as only the eggs (of an ascarid and a trematode) were found in some samples. From the organisms identified, the ectoparasites (lice-Struthiolipeurus rheae, S. nandu; mites-Dermoglyphus pachycnemis, Gabucinia bicaudata), helminths (cestoda-Houttuynia struthionis- and nematoda-Libyostrongylus sp., Codiostomum struthionis-) and the ciliate Balantidium struthionis are known as ratite specific parasites. Capillaria eggs and larvae were also found; there are no previous records of this parasite from ostriches, and the data available do not allow to do a temptative specific diagnosis. Among protozoa, most of the species now found are described for the first time in ratites. They include organisms also found in other birds (Trichomonas gallinae, Tetratrichomonas gallinarum, Chilomastix gallinarum, Spironucleus meleagridis and Pleuromonas jaculans), and organisms whose specific status cannot be established until further analysis are performed (Cryptosporidium sp., Eimeria sp. and/or Isospora sp., Entamoeba sp. of the one-nucleate and of the eight-nucleate mature cyst groups, Endolimax sp., Iodamoeba sp., Monocercomonas sp., Retortamonas sp., Giardia sp., Blastocystis sp. and euglenids).     

Intestinal parasites of baboons (Papio cynocephalus anubis) and chimpanzees (Pan troglodytes) in Gombe National Park.

A survey of gastrointestinal parasites of nonhuman primates in Gombe National Park, Tanzania, was conducted during July and August 1989. Fresh fecal samples collected from 35 baboons (Papio cynocephalus anubis) and 20 chimpanzees (Pan troglodytes) were analyzed for intestinal parasites. Parasite identifications were based upon the size and appearance of eggs and larvae on both direct and fecal flotation examinations. Seven species of helminths and three species of protozoa were noted in the chimpanzees, and seven species of helminths, two species of trematodes, and one species of protozoa were noted in baboons. An unidentified parasite, with a thick egg shell similar to that of ascarids, was noted in both the chimpanzee and baboon population. Unfortunately a definitive identification of this parasite could not be made based upon egg size and morphology alone and insufficient numbers of larva were present to aid identification. One previously unreported parasite, Schistosoma mansoni, was identified in the baboon population. This is the first report of a schistosome in Gombe's nonhuman primate population.     

Gastrointestinal parasites of mountain gorillas (Gorilla gorilla beringei) in the Parc National des Volcans, Rwanda.

Ninety-eight fecal samples were collected from 74 free-living mountain gorillas (Gorilla gorilla beringei) from the Parc National des Volcans, Rwanda, between July 1995 and January 1997 and examined for parasites by Sheather's sugar and zinc sulfate flotation methods, trichrome staining, and larval cultures. All samples contained at least one parasite. Seventeen endoparasites were identified, including eight protozoa, seven nematodes, one cestode, and one trematode. Two species of arthropod mite were also recovered from the fecal samples. Parasites observed on fecal examinations included strongyle/trichostrongyle-type eggs (72/74) (representing Oesphagostomum sp., Trichostrongylus sp., Hyostrongylus spp., and possibly Murshidia sp.), Strongyloides sp. (1/74), Trichuris trichiura (2/74), Probstmayria sp. (7/74), Anoplocephala sp. (63/74), Entamoeba hartmanni cysts and trophozoites (19/70), Endolimax nana cysts (31/70), Iodamoeba buetschlii cysts (11/70), Endolimax nana or Iodamoeba buetschlii trophozoites (63/70). Entamoeba coli cysts and trophozoites (14/70), Entamoeba histolytica trophozoite (1/70), Chilomastix sp. cysts and trophozoites (31/70), and Giardia sp. cysts (2/70). In addition, one ascarid and one trematode egg were seen. There were no significant differences in the prevalence of parasites between males and females and between age groups: however, infants and juveniles appeared to have a lower prevalence of Anoplocephala gorillae, and the silverbacked males appeared to have a higher prevalence of Probstmayria sp. Parasite prevalence was consistent among the five social groups studied except Susa group had a significantly lower prevalence of Anoplocephala gorillae. Trichuris trichiura, Strongyloides sp., Chilomastix sp., and Endolimax nana were identified for the first time in this population, and it is possible that these parasites were of human origin. Although there were no obvious clinical effects due to the presence of these parasites, six parasites identified (Trichuris trichiura, Strongyloides sp., Oesphagostomum sp., Trichostrongylus sp., Entamoeba histolytica, and Giardia sp.) could potentially be pathogenic. Some of the parasite products and cultured larvae could not be speciated.     

Efficiency of feeding Duddingtonia flagrans chlamydospores to grazing ewes on reducing availability of parasitic nematode larvae on pasture

Gastrointestinal nematodes are of concern in sheep production because of production and economic losses. Control of these nematodes is primarily based on the use of anthelmintic treatment and pasture management. The almost exclusive use of anthelmintic treatment has resulted in development of anthelmintic resistance which has led to the need for other parasite control options to be explored. The blood sucking abomasal parasitic nematode Haemonchus contortus causes severe losses in small ruminant production in the warm, humid sub-tropic and tropics. This study evaluated the effectiveness of a nematode trapping fungus, Duddingtonia flagrans, in reducing availability of parasitic nematode larvae, specifically H. contortus, on pasture. Chlamydospores of D. flagrans were mixed with a supplement feed which was fed daily to a group of crossbred ewes for the duration of the summer grazing season. A control group was fed the same supplement feed without chlamydospores. A reduction in infective larval numbers was observed in fecal cultures of the fungus-fed group. Herbage samples from the pasture grazed by the fungus-fed group also showed a reduction in infective larvae. There were no significant (P > 0.05) differences in overall fecal egg count, packed cell volume or animal weight between fungus-fed and control groups. Tracer animals were placed on the study pastures at the end of the study to assess pasture infectivity. Although tracer animals were only two per group, those that grazed with the fungus-fed group had substantially reduced (96.8%) nematode burdens as compared to those from the control group pasture. Results demonstrated that the fungus did have activity against nematode larvae in the feces which reduced pasture infectivity and subsequently nematode burdens in tracer animals. This study showed that D. flagrans, fed daily to grazing ewes, was an effective biological control agent in reducing a predominantly H. contortus larval population on pasture.     

Efficacy of an Arthrobotrys oligospora N mutant in nematode-trapping larvae after passage through the digestive tract of sheep

For oral applications, biocontrol of animal parasitic nematodes in ruminants, ion beam implanted nematode-trapping fungi must have the capacity to survive the passage through the digestive tract and be efficient in reducing infective larvae of nematodes in the faeces.Ion beams induced mutation in the spores of nematode-trapping Arthrobotrys oligospora. Mutants with genetic stability were bred. The fungi were cultured in bottles with corn kernels as growth media, and spores of different doses were, respectively, administered orally to each group of sheep naturally infected with gastrointestinal nematodes. The control group did not receive fungi. The faeces of these experimental animals were collected and faecal cultivations carried out. The fungal germination, growth, reproduction and predation of livestock parasitic nematode larvae were tested in laboratory. The efficacy of an A. oligospora N mutant in nematode-trapping larvae after passage through the digestive tract of sheep was tested. The results indicated that the ion beam implantation induced mutation of the nematode-trapping fungi is a positive mutation. The mutant spores through the digestive tract of sheep can kill livestock parasite nematode larvae in vitro. These results indicate the potential of the A. oligospora N mutant as a biological control agent for sheep nematodes.This study showed that such biotechnology could be explored for improving the effectiveness of the use of fungal infections to control livestock parasitic nematodes. This work represents the first application of nematode-trapping fungi in eukaryotic microorganisms.     

Induction of nematode-trapping organs in the predacious fungus Arthrobotrys oligospora (Hyphomycetales) by infective larvae of Ostertagia ostertagi (Trichostrongylidae)

Laboratory experiments were designed to study the influence of temperature, concentrations of nematodes, oxygen tension, light, and nutrient levels, on the induction of nematode-trapping hyphal nets in the predacious fungus Arthrobotrys oligospora. When induced by infective Ostertagia ostertagi larvae, a maximum number of nets was produced at 20 degrees C, at which temperature nets in surplus were produced at larval concentrations up to 1,000 larvae per cm2. A. oligospora did not produce nets in an anaerobic atmosphere containing 21% CO2 (v/v), and net induction was suppressed to a certain degree by exposure to light. The composition of the medium had an important influence on the saprophytic growth and the net-forming capability of A. oligospora as a maximum number of nets was induced at a relatively low concentration of corn meal supporting the relatively sparse mycelium. It was shown that a proportion of trapping nets in A. oligospora maintained their trapping potential for more than 7 weeks when the temperature was below 25 degrees C. Induction of nematode-trapping organs in A. oligospora is discussed in relation to control of infective nematode parasite larvae in cow pats.     

The experimental establishment of ruminant nematodes in European hares (Lepus europaeus)

The factors that control the demography of European hare Lepus europaeus populations are poorly understood, but it has been recognized that the decline of hares in Europe is associated with an increasing intensity of agricultural activity. Many mechanisms have been suggested. We propose another mechanism; a negative impact arising from ingestion of the infective larvae of ruminant livestock. We dosed juvenile hares from a worm-free colony with a conservative dose from a mixed culture of infective larvae of the nematode parasites of sheep and cattle. We examined the hares post-mortem for the establishment of those ruminant nematodes, differences in weight changes, and the shedding of eggs.     

Efficacy of albendazole and moxidectin and resistance to ivermectin against Libyostrongylus douglassii and Libyostrongylus dentatus in ostriches

Anthelmintic resistance has emerged globally as a problem amongst nematode of livestock and has been particularly well documented in equine and small ruminants. There are no studies regarding the efficacy of anthelmintics against the hematophagous nematodes in ostriches, Libyostrongylus dentatus; and just a few on L. douglassii. Here the efficacy of albendazole, ivermectin and moxidectin were evaluated against these two species in an ostrich farm in Minas Gerais state, Brazil. The feces were collected on the day of treatment and after 13days of an oral dose of albendazole (6mg/kg), or an injected dose (0.2mg/kg) of ivermectin or moxidectin. The fecal egg count reduction test and coprocultures were performed to determine possible resistance against the drugs used. An efficacy of 60% was found for ivermectin, while albendazole and moxidectin were 100% effective. Both worm species appeared to have reduced sensitivity to ivermectin.     

The effects of birdsfoot trefoil (Lotus corniculatus) and chicory (Cichorium intybus) when compared with perennial ryegrass (Lolium perenne) on ovine gastrointestinal parasite development, survival and migration

Two experiments were conducted to investigate the effects of birdsfoot trefoil and chicory on parasitic nematode development, survival and migration when compared with perennial ryegrass. In experiment one, sheep faeces, containing 10,385 Cooperia curticei eggs were added to 25 cm diameter pots containing birdsfoot trefoil, chicory or ryegrass, and the pots maintained under optimal conditions for nematode parasite development. Replicate pots of each forage type were destructively sampled on day 8, 16, 20, 28 and 37 to collect the nematode larvae. When forages were compared on a dry matter basis, by day 16 there were 31% and 19% fewer larvae on birdsfoot trefoil and chicory than on ryegrass, respectively (P<0.01). In the second experiment, replicate 1m(2) field plots of birdsfoot trefoil, chicory and ryegrass were sub-sampled on day 14, 21, 35 and 49 for larval counts following the application of sheep faeces containing 585,000 Teladorsagia circumcincta eggs to each plot on day 0. Results showed there were a minimum of 58% and 63% fewer infective stage parasitic larvae on birdsfoot trefoil and chicory, respectively, compared with ryegrass on day 14 and 35 when forages were compared on a forage dry matter, plot area sampled and leaf area basis (P<0.01). Overall, these results indicate that the number of infective stage larvae on birdsfoot trefoil and chicory pasture was reduced by the effect of their sward structure on the development/survival/migration of ovine parasitic nematodes. These effects may be one of the ways in which these forages may affect parasitic infections in grazing livestock.     

Gastrointestinal nematodes of small ruminants: life cycle, anthelmintics, and diagnosis.

Strongylid gastrointestinal nematodes are an important cause of disease and economic loss in small ruminants. These parasites are particularly important in the eastern United States where the bloodsucking parasite Haemonchus contortus is the predominant species. Sheep and goats are infected with gastrointestinal nematodes while grazing and the biology of the infective larvae on the pastures is important in planning effective control programs. Fecal egg counts can be used to monitor control programs and drug efficacy and evaluate levels of pasture contamination.     

The epidemiology of nematode infections of sheep

Nematode parasites have been a major factor limiting sheep production in New Zealand for more than 100 years. Twenty-nine species of nematodes were unintentionally introduced with sheep into New Zealand, but it is principally species of Haemonchus, Ostertagia, Trichostrongylus, Nematodirus and Cooperia that are associated with production losses and clinical disease. The seasonal dynamics of nematode infection are the consequence of complex inter-relationships between the sheep, their husbandry and the prevailing climate. The patterns of pasture contamination by nematode eggs and then larvae and the subsequent levels of infection in ewes and lambs are broadly similar throughout New Zealand. Numbers of infective larvae on pasture build up over summer to a peak in autumn/early winter with, in some years, a spring peak derived from the parturient rise in faecal nematode egg counts (FEC), expressed in eggs per gram of faeces (epg), in lactating ewes. The immune capability of lambs is initially low but increases with the magnitude and duration of exposure to infection. Once significant immunity has developed (usually by 10-12 months of age), sheep are capable of markedly restricting parasite infection, except during times of disease, malnutrition or stress. For the effective control of nematode parasites, farmers have come to rely almost exclusively on broad-spectrum anthelmintics. However, issues relating to resistance, residues and eco-toxicity increasingly threaten the sustainability of chemotherapy. In order to maintain present levels of parasite control and productivity in the long term, farmers need to integrate management practices aimed at minimising animal exposure to parasites with reduced reliance on anthelmintics.