Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems.

Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump's bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.

Multiparasitism in a wild cat (Leopardus colocolo) (Carnivora: Felidae) in southern Brazil.

Parasitic diseases reflect the health and balance of ecosystems, affecting not only individuals but also entire populations or communities. The aim of this study was to report on the diversity of parasitic helminths detected in the feces of a wild feline in southern Brazil. Parasites were obtained from fecal samples, and four techniques were used for parasitological examination: direct examination, centrifugal flotation with zinc sulfate (Faust technique), simple sedimentation (Hoffman technique) and Baermann-Moraes. The parasites were identified through micrometry and morphology, as follows: Ancylostoma sp., Toxocara sp., Trichuridae, Aelurostrongylus abstrusus, Alaria sp., and Spirometra sp. We recorded the genus Ancylostoma parasitizing L. colocolo for the first time.

Multiparasitism in a wild cat (Leopardus colocolo) (Carnivora: Felidae) in southern Brazil / Multiparasitismo em um felino silvestre (Leopardus colocolo) (Carnivora: Felidae) no Sul do Brasil

Abstract Parasitic diseases reflect the health and balance of ecosystems, affecting not only individuals but also entire populations or communities. The aim of this study was to report on the diversity of parasitic helminths detected in the feces of a wild feline in southern Brazil. Parasites were obtained from fecal samples, and four techniques were used for parasitological examination: direct examination, centrifugal flotation with zinc sulfate (Faust technique), simple sedimentation (Hoffman technique) and Baermann-Moraes. The parasites were identified through micrometry and morphology, as follows: Ancylostoma sp., Toxocara sp., Trichuridae, Aelurostrongylus abstrusus, Alaria sp., and Spirometra sp. We recorded the genus Ancylostoma parasitizing L. colocolo for the first time.

Resumo Doenças parasitárias refletem a saúde e o equilíbrio dos ecossistemas, influenciando não só um indivíduo e sim uma população ou comunidade. Este trabalho teve por objetivo relatar a diversidade de helmintos encontradas nas fezes de um felino silvestre na região Sul do Brasil. Os parasitos foram obtidos a partir de amostras fecais, sendo utilizadas quatro técnicas para os exames parasitológicos: exame direto, centrífugo-flutuação com sulfato de zinco (Técnica de Faust), sedimentação simples (Técnica de Hoffman) e Baermann-Moraes. Os parasitos foram identificados através de micrometria e morfologia, sendo esses: Ancylostoma sp., Toxocara sp., Trichuridae, Aelurostrongylus abstrusus, Alaria sp. e Spirometra sp. Estudos da fauna parasitária de animais silvestres são relevantes, tanto para o equilíbrio e saúde desses animais, como para o controle e prevenção de doenças transmitidas ao homem. Ancylostoma spp. foi identificado pela primeira vez em L. colocolo.

Baruscapillaria obsignata: a serious cause of enteropathy and high mortality in turkeys (meleagris gallopavo).

BACKGROUND: Capillariasis, an important parasitic disease of birds is caused at least by seven different genera of trichurid nematodes with clinical outcome ranging from mild enteritis to high mortality. OBJECTIVE: This study was aimed to investigate the causative agent involved in high mortality associated with severe enteric illness among turkey flocks in an organized commercial poultry farm at Bareilly, India. MATERIALS AND METHODS: Turkey carcasses (n = 119) and fecal samples from the affected deep litter pen constituted as the study materials. The disease was investigated by systematic necropsy, direct microscopy and histopathology. Representative samples were screened for other enteric pathogens. RESULTS: Microscopic examination of mucosal scraping revealed capillarid worms and their eggs in all the samples. The morphological features of adult worms were typically consistent to Baruscapillaria obsignata. Histopathology exhibited thickened muscular and mucosal layers, mononuclear and heterophilic infiltration in the lamina propria, blunting and clubbing of villi, epithelial denudation and sections of capillarid worms. Administration of levamisole at 80 ppm in drinking water reduced the mortality, clinical illness and worm load after three days of therapy. CONCLUSIONS: The capillarid worms in different avian hosts can cause different clinical manifestations and outcomes. From India, this is the first report describing intestinal pathology caused by B. obsignata in turkeys. We conclude that the B. obsignata infection is capable of causing life threatening enteropathy in turkeys and, hence, routine screening, scheduled deworming and good litter management are crucial to control the infection and its associated loss.

Three-Dimensional Screen: A Comprehensive Approach to the Health Monitoring of Zebrafish.

Considering the numbers of zebrafish held in the laboratories, it is relevant to develop some tools to monitor the health of the animals, as well as their biotope. Environmental samples can be used to detect aquatic pathogens. Comprehensive health monitoring would thus seek pathogens in three dimensions of the animals and microbes' habitat: the fish, the sludge, and the water. This three-dimensional approach is called the 3D screen and it introduces some complementary tools to routine sentinel screening. For example, sludge and sump swabs analyses allow an efficient detection of pathogens at a low cost and with a fast turnover. These assays are particularly useful in cases of Pseudocapillaria tomentosa infestation or Mycobacterium haemophilum outbreak. Indeed, such a broader choice of diagnostic tests gives flexibility for the veterinarian to investigate Mycobacterium spp. presence in the water systems and fish colonies. Some other robust additional analysis, like the mortality rate monitoring, quickens the decision-making process. The 3D screen describes how this new toolbox can be used efficiently to monitor laboratory fish health.

A simple and efficient protocol for the treatment of zebrafish colonies infected with parasitic nematodes.

Abstract Our zebrafish colony experienced a period of increased mortality rate of 6.5 times more deaths per month in a colony of over 13,000 zebrafish (Danio rerio), which developed over 3 months. We observed that before death, affected fish appeared emaciated, often with an abdominal bulge. We performed dissection on 18 fish that had this appearance and found in 15 that their gut was infected with a nematode that closely resembled Pseudocapillaria tomentosa. We devised a treatment protocol for this nematode infection, which involved addition of fenbendazole, a drug used to treat nematode infections in cattle and sheep, to the fish feed. Fenbendazole produced no severe side effects in the fish and several treatments have effectively eradicated the parasite from our colony. The mortality rate of our fish has decreased to a value of 0.7%/month (p<0.001, equal to that before the infection). We propose this protocol as an inexpensive alternative to having to rederive an entire colony from bleached eggs, and as a prophylactic measure used in quarantine facilities on a regular basis.

First report of Eucoleus boehmi (syn. Capillaria boehmi) in dogs in north-western Italy, with scanning electron microscopy of the eggs.

Dogs can be infected by several nematodes of the Trichuridae family. Trichuridae eggs are all similar, barrel shaped with polar plugs, and misdiagnosis among different species can occur. The most common species is Trichuris vulpis, while the respiratory parasites Eucoleus boehmi (syn. Capillaria boehmi) and Eucoleus aerophilus (syn. Capillaria aerophila) are rarely observed in pets. E. boehmi is reported for the first time in this study in north-western Italy with other Trichuridae. Dog faecal samples (270) were examined by flotation. E. boehmi (2.2%), E. aerophilus (4.4%) and T. vulpis (12.2%) were found; identification was done with measurements and through observation of morphological characters already known. The specific identification of E. boehmi was confirmed using scanning electron microscopy: its egg shell shows a dense network with a fine mesh, surrounding small pits, on the contrary E. aerophilus eggs present a thick mesh with wide depressions, while T. vulpis eggs surface is smooth.

Survey on helminthofauna in pheasants from Eastern Europe.

In order to assess the parasitological status in imported pheasants, 51 birds (Phasianus colchicus) coming from Poland and Rumania and used in Italy in repopulation interventions for hunting purposes were examined. From each animal the trachea, oesophagus, crop and intestine were collected and examined for the presence of nematodes. The examination of the oesophagi and crops of 5 birds revealed the presence of parasites pertaining to the family Capillariidae: Eucoleus contortus was found in all of the 5 animals, E. annulatus was present along with the previous parasite in one animal coming from Poland. The examination of the tracheae revealed the presence of Syngamus trachea in 5 animals (9.80%). Adult or larval stages of Heterakis gallinarum (37.25% of birds) and Capillariidae (35.29%) were found in the intestinal tracts. Aonchotheca caudinflata was detected only in one bird coming from Poland; the capillarids found in all of the remaining pheasants exhibited morphological characteristics referable to Capillaria phasianina, a species never reported in Italy. The release of game from foreign countries, therefore, may always constitute a risk for the autochthonous one due to the spread of new parasitic infections.

Trichurid nematodes in ring-necked pheasants from backyard flocks of the State of Rio de Janeiro, Brazil: frequency and pathology.

The present investigation is related to the frequency of infection and to the gross and microscopic lesions associated to the presence of trichurid worms in 50 ring-necked pheasants (Phasianus colchicus) from backyard flocks in the state of Rio de Janeiro, Brazil. In the investigated birds, the overall infection rate was of 74%, with the presence of Eucoleus perforans with 72% of prevalence and 21.2 of mean intensity, in the esophageal and crop mucosa and rarely in the junction of the proventriculus and esophagus, E. annulatus with 2% and 3 in the crop mucosa, Capillaria phasianina, with 12% and 4.3 in the cecum and small intestine and Baruscapillaria obsignata, for the first time referred in this host, with 2% and 1 in the small intestine. Clinical signs were absent. The gross lesions observed in the crop and esophagus of 14 (38.9%) pheasants parasitized with E. perforans were thickening, small nodules, congestion, and petechial haemorrhages in the mucosa. These birds presented a mean infection of 37.5 and a range of infection of 10-82. The microscopic lesions revealed chronic esophagitis with diffuse inflammatory process in the lamina propria characterized mostly by a mononuclear cell infiltrate and also with the presence of granulocytes. In the case of the parasitism of pheasants with C. phasianina, the gross lesions were absent; microscopic lesions were characterized by chronic typhlitis with mononuclear infiltrate. Gross and microscopic lesions were absent in the pheasants parasitized with E. annulatus and B. obsignata.

Histological study of Trichosomoides nasalis (Nematoda: Trichinelloidea) in the nasal cavities of the murid Arvicanthis niloticus, with associated pathology.

Histological study of the nasal cavities and upper maxillae of Arvicanthis niloticus naturally infected with Trichosomoides nasalis shows that the female worms reside in the epithelial monolayer of the nasal mucosa of the posterior and median cavities. Eggs laid by T. nasalis were infiltrated between the female body wall and the epithelial lining. Small groups of eggs, mixed with mucus and polymorphonuclear cells, were found in the nasal lumen, freed by rupture of the stretched epithelium. Two females and a few eggs were also found in the connective tissues. One male was found in a female uterus and two were apparently in the lumen of the nasal cavity but the surrounding tissues were disrupted. No male was identified in the lamina propria of the mucosa. However, significant inflammatory lesions occurred in the lamina propria, similar to those induced by the males of Anatrichosoma spp. which live in this part of the mucosa. In rodents, the lesions resulted in rhinosinusitis characterised by a lymphocytic infiltration leading to nasal obstruction.

Pseudocapillaria (Ichthyocapillaria) ophisterni sp. n. (Nematoda : Capillariidae) from the swamp-eel Ophisternon aenigmaticum (Pisces) in Mexico.

A new nematode species, Pseudocapillaria ophisterni sp. n., is described from the intestine and rarely from the stomach of the swamp-eel, Ophisternon aenigmaticum Rosen et Greenwood, from Catemaco Lake, Veracruz, Mexico. In having both caudal lobes in the male interconnected by a distinct dorsal membrane, it belongs to the subgenus Ichthyocapillaria. It differs from the three species in this subgenus mainly in possessing either a distinctly longer spicule or a smaller length of oesophagus relative to body length. It also differs in host type and geographical distribution. P. ophisterni is the first capillariid species reported from synbranchiform fishes.

[Nematode parasites of birds of the fauna in Tunisia]. / Nématodes parasites d'oiseaux de la faune de Tunisie.

330 birds of Tunisia were necropsied; they belong to 73 species among 29 families and 13 orders. We so collect 36 species of nematodes from 9 families. The relative importance of these is variable. The best represented are: Capillariidae (6 species), Spiruridae (6 species), Acuariidae (9 species) and Filariidae (6 species). The parasitism by nematodes is not uniform. Among the 330 birds autopsied only 51 were parasited by nematodes (15.45%) among 25 of the examined birds species (34.2%). Among these 25, sixteen (64%) presented only one species of parasitic nematode, six (24%) arboured two, (8%) three and only one, (partridges) presents six species of nematodes.

Capillaria procyonis (Nematoda: Trichuroidea) eggs from the tongue of the raccoon (Procyon lotor).

During an investigation to determine the prevalence and intensity of larvae of Trichinella spiralis in raccoons (Procyon lotor), unembryonated capillariid-type eggs were found routinely in the sediment resulting from 1% pepsin-HCl acid digests of the tongue. The sediment from 36 (90%) of the 40 tongue digests examined contained these capillariid-type eggs. Mean measurements of egg width and length were 25 +/- 2 microns and 62 +/- 2 microns, respectively. In addition, the surface of the egg shell had a pitted appearance. Adult parasites were not found in any of the tongue digests. The egg measurements and surface topography agree with what has been described previously and are tentatively identified as Capillaria procyonis. The detection of C. procyonis eggs in the present report indicates that this parasite inhabits the epithelial lining of the raccoon tongue, in addition to the esophagus, and that eggs expelled by females are deposited in the tracts these parasites create as they migrate.

Anatrichosoma haycocki sp. n. (Nematoda: Trichuridae) from the paracloacal glands of Antechinus spp. With notes on Skrjabinocapillaria skarbilovitsch.

Anatrichosoma haycocki sp. n. (Nematoda: Trichuridae) is described from the paracloacal glands of Antechinus swainsonii (Waterhouse, 1840) and A. stuartii Macleay, 1841 (Marsupialia: Dasyuridae) from Nadgee State Forest, New South Wales, Australia. Immature male and female worms occur free in the intestine. A. haycocki is distinguished from other species of Anatrichosoma by the following suite of morphological characters: small size, absence of longitudinal cuticular striations in both sexes, subterminal constriction of male tail bearing 14 minute papillae and inflation of posterior portion of mature female giving Trichuris-like appearance. It is also characterised by its occurrence in paracloacal glands in a genus of small carnivorous dasyurid marsupial restricted to Australia and New Guinea. Skrjabinocapillaria rodentium Wertheim and Chabaud, 1979 is recognised as a synonym of Anatrichosoma gerbillis (Bernard, 1964). The genus Skrjabinocapillaria Skarbilovitsch, 1946 is placed as a synonym of Capillaria Zeder, 1800, resulting in the new combination for the type species: C. eubarsata (Skarbilovitsch, 1946) comb. nov. It is suggested that deep penetration of the female uterus by the male at insemination is a behavioural feature common to members of the Trichinelloidea and that the morphology of the male reproductive tract reflects the mechanisms of penetration employed by members of each genus.

An epidemiological survey of wild caught and domestic born rhesus monkeys (Macaca mulatta) for Anatrichosoma (Nematoda: Trichinellida).

A group of wild caught rhesus monkeys and their domestic born infants were examined for Anatrichosoma, a nematode that inhabits the nasal mucosa and subcutaneous tissue of the face, hands, and feet. The diagnosis was made using nasal swabs. Fifty-four percent of the wild caught animals reported positive ina survey taken 3 years earlier were negative based upon results of reexamination, while one animal was found positive that had been reported negative. None of the infants examined had positive samples for Anatrichosoma.