Fucose Ameliorates Tritrichomonas sp.-Associated Illness in Antibiotic-Treated Muc2-/- Mice.

The mucus layer in the intestine plays a critical role in regulation of host-microbe interactions and maintaining homeostasis. Disruptions of the mucus layer due to genetic, environmental, or immune factors may lead to inflammatory bowel diseases (IBD). IBD frequently are accompanied with infections, and therefore are treated with antibiotics. Hence, it is important to evaluate risks of antibiotic treatment in individuals with vulnerable gut barrier and chronic inflammation. Mice with a knockout of the Muc2 gene, encoding the main glycoprotein component of the mucus, demonstrate a close contact of the microbes with the gut epithelium which leads to chronic inflammation resembling IBD. Here we demonstrate that the Muc2-/- mice harboring a gut protozoan infection Tritrichomonas sp. are susceptible to an antibiotic-induced depletion of the bacterial microbiota. Suppression of the protozoan infection with efficient metronidazole dosage or L-fucose administration resulted in amelioration of an illness observed in antibiotic-treated Muc2-/- mice. Fucose is a monosaccharide presented abundantly in gut glycoproteins, including Mucin2, and is known to be involved in host-microbe interactions, in particular in microbe adhesion. We suppose that further investigation of the role of fucose in protozoan adhesion to host cells may be of great value.

Malaria parasites and related haemosporidians cause mortality in cranes: a study on the parasites diversity, prevalence and distribution in Beijing Zoo.

BACKGROUND: Malaria parasites and related haemosporidian parasites are widespread and may cause severe diseases in birds. These pathogens should be considered in projects aiming breeding of birds for purposes of sustained ex situ conservation. Cranes are the 'flagship species' for health assessment of wetland ecosystems, and the majority of species are endangered. Malaria parasites and other haemosporidians have been reported in cranes, but the host-parasite relationships remain insufficiently understood. Morbidity of cranes due to malaria has been reported in Beijing Zoo. This study report prevalence, diversity and distribution of malaria parasites and related haemosporidians in cranes in Beijing Zoo and suggest simple measures to protect vulnerable individuals. METHODS: In all, 123 cranes (62 adults and 61 juveniles) belonging to 10 species were examined using PCR-based testing and microscopic examination of blood samples collected in 2007-2014. All birds were maintained in open-air aviaries, except for 19 chicks that were raised in a greenhouse with the aim to protect them from bites of blood-sucking insects. Bayesian phylogenetic analysis was used to identify the closely related avian haemosporidian parasites. RESULTS: Species of Plasmodium (5 lineages), Haemoproteus (1) and Leucocytozoon (2) were reported. Malaria parasites predominated (83% of all reported infections). The overall prevalence of haemosporidians in juveniles was approximately seven-fold higher than in adults, indicating high susceptibility of chicks and local transmission. Juvenile and adult birds hosted different lineages of Plasmodium, indicating that chicks got infection from non-parent birds. Plasmodium relictum (pSGS1) was the most prevalent malaria parasite. Mortality was not reported in adults, but 53% of infected chicks died, with reports of co-infection with Plasmodium and Leucocytozoon species. All chicks maintained in the greenhouse were non-infected and survived. Species of Leucocytozoon were undetectable by commonly used PCR protocol, but readily visible in blood films. CONCLUSION: Crane chicks often die due to malaria and Leucocytozoon infections, which they likely gain from wild free-living birds in Beijing Zoo. Molecular diagnostics of crane Leucocytozoon parasites needs improvement. Because the reported infections are mainly chick diseases, the authors recommend maintaining of juvenile birds in vector-free facilities until the age of approximately 6 months before they are placed in open-air aviaries.

Experimental infections with the protozoan parasite Histomonas meleagridis: a review.

In recent years, a number of studies about Histomonas meleagridis, and more specifically about experiments in vivo involving H. meleagridis to investigate the pathogenicity and efficacy of drugs or vaccines, have been published. Together with older publications, a considerable amount of information about experimental infections with H. meleagridis exist, which is helpful for planning future animal studies and can reduce the number of birds used in such studies toward better animal welfare. One hundred sixty-seven publications describing experimental infections with H. meleagridis were published in scientific journals between 1920 and 2012. One hundred forty-two of these publications describe infections of turkeys (Meleagris gallopavo) and 52 infections of chickens (Gallus gallus). In 18 studies, experiments involving other species were done. The most popular routes of infection were the intracloacal application of histomonal trophozoites from culture material, from lesions or from feces of infected birds, or using larvae of the cecal worm Heterakis gallinarum (83 studies) and the oral application of eggs or other stages of the cecal worm containing histomonal stages (83 studies). During the last 10 years, intracloacal application of trophozoites has become the most popular way to experimentally infect birds with H. meleagridis due to its high reproducibility and reliability. In most studies, infection doses of several 10,000 or 100,000 histomonal trophozoites were used for infection, and the resulting mortality in turkeys was more than 70 %. First mortality can occur as early as 6 days p.i.; peak mortality usually is 13-15 days p.i. Lower infection doses may delay mortality about 2 days. In chickens infected by the intracloacal route, mortality and clinical signs are rare, but infection rates are similar. Cecal lesions can be observed from 3 to 4 days p.i., lesions up to 3 weeks p.i.; liver lesions may be lacking completely or be present only in a small number of birds. In most studies infecting birds with Heterakis eggs containing histomonal stages, several 100 to 1,000 Heterakis eggs were used. However, lower doses might be sufficient, as infection with as few as 58 eggs per bird caused a mortality up to 90 % in turkeys. Clinical symptoms start 9 days p.i., and first mortality occurs after 12 days, while most of the infected birds die between 19 and 21 days p.i. The infectivity of Heterakis eggs containing histomonal stages for chickens is similar as for turkeys, but mortality and clinical signs are rare. Further infection was done by oral application of histomonal trophozoites either grown in culture or using lesions or feces of infected birds (26 studies). These yielded very mixed results, with infection rates between 0 and more than 80 % in turkeys and chickens. After successful oral infection of turkeys, mortality occurs at roughly the same time as after intracloacal infection. Further 18 studies employed seeder birds to infect in-contact birds. Other means of infection were exposure to contaminated soil or litter (22 studies), feeding contaminated earthworms (7 studies), intracecal inoculation (4 studies), or parenteral injection (4 studies). Main methods to assess the course of the infection were mortality, observation of clinical signs and pathological lesions, monitoring of the weight of the infected birds, and detection of the parasite by various methods.

Systemic histomoniasis associated with high mortality and unusual lesions in the bursa of Fabricius, kidneys, and lungs in commercial turkeys.

An outbreak of systemic histomoniasis in 9-to-11-wk-old meat-type male turkeys associated with high mortality is reported. Clinical signs in turkeys included anorexia, depression, diarrhea, loss of weight, and increased mortality between 7 and 13 wk of age. Seven-week cumulative mortality in the turkeys of affected houses ranged from 24% in one to 68% in the other compared with 1.3% of turkeys in the house not affected by histomoniasis. Necropsy of the turkeys revealed enlargement of the livers, most of which had numerous pale white nodules ranging in size from 0.3 to 1.5 cm in diameter. Cecal walls were severely thickened, the lumens were distended with caseous cores, and the mucosa was ulcerated. The bursa of Fabricius in four birds was enlarged and contained pale yellow exudate in their lumens. The kidneys, pancreas, and spleens in a few birds had pale or yellow foci. Microscopically, there were multifocal necrosis and granulomatous inflammation in the liver, ceca, peritoneum, bursa of Fabricius, kidneys, lungs, pancreas, proventriculus, and in spleens associated with histomonads. Electron microscopy also confirmed the presence of histomonads in the liver and ceca. This is the first report of naturally occurring systemic histomoniasis in turkeys affecting the bursa of Fabricius, kidneys, lungs, pancreas, and proventriculus.

Intestinal pathogen clearance in children with severe acute malnutrition is unrelated to inpatient morbidity.

BACKGROUND & AIMS: Children with Severe Acute Malnutrition (SAM) often suffer from diarrhea, which is associated with increased mortality. The contribution of intestinal bacteria, parasites and viruses to morbidity such as diarrhea in SAM remains poorly understood. To evaluate their association with clinical outcomes, we detected stool pathogens in children with SAM at hospital admission and after clinical stabilization prior to discharge. METHODS: 15 intestinal pathogens, fecal calprotectin and C-reactive protein (CRP) were determined at admission and after clinical stabilization in children aged 8-59 months (n = 47) hospitalized in Malawi for complicated SAM. Differences in fecal pathogens, intestinal and systemic inflammation, and clinical outcomes between time points were evaluated using the Wilcoxon Signed-Rank test or Wilcoxon rank-sum test. RESULTS: On admission pathogens were present in nearly all children and after clinical stabilization many were cleared with only 55% of children still harboring a pathogen (89% vs. 55%, p = 0.001). Nosocomial infections were infrequent. The pathogens Giardia lamblia and Shigella spp. were most likely to persist. After clinical stabilization, fecal calprotectin was higher in children harboring a pathogen (median (IQR): 383 mg/kg (903-149 mg/kg) vs 140 mg/kg (300-71 mg/kg), p = 0.03). CRP did not correlate with fecal calprotectin levels nor was it associated with pathogen detection. Presence of stool pathogens was not associated with clinical outcomes such as diarrhea. CONCLUSIONS: Fecal pathogens were very common and cleared in most children with complicated SAM treated with antibiotics. The presence of stool pathogens after stabilization was associated with increased intestinal inflammation but not with clinical outcomes. (http://www.isrctn.com/ISRCTN13916953).

Mitigation of lethal effects of Karlodinium veneficum and K. armiger on Sparus aurata: changes in haematocrit and plasma osmolality.

Between January and April 2000, several experiments were performed during a Karlodinium spp. proliferation in Alfacs Bay (Ebro delta, NW Mediterranean) to determine the effects of these dinoflagellates on sea bream Sparus aurata cultivated in the area. Moribund fish showed an increase in plasma osmolality together with a decrease in the haematocrit percentage compared to control fish. The efficacy of copper sulphate, hydrogen peroxide, potassium permanganate and formalin against Karlodinium spp. was also tested. None of these treatments had mitigation effects when applied in the presence of fish; on the contrary, lethal effects appeared at lower Karlodinium spp. densities compared to fish control groups. When a lytic agent, such as copper sulphate, was used as a water pre-treatment, in the absence of fish, Karlodinium spp. toxicity was significantly reduced. Protocols for water pre-treatments were studied as a potential tool for combating Karlodinium spp. in fish farms.

Incidence of histomonosis in turkeys in France since the bans of dimetridazole and nifursol.

Between April 2003 and March 2005, 113 outbreaks of histomonosis were recorded in standard turkey farms in France, and 15 cases were recorded in turkey breeding centres. Most of the cases were in north-west France, the principal farming area for turkeys. The majority of the cases occurred during the hottest months, from April to September. Large numbers of cases occurred among birds from four to eight weeks of age, but there were some cases in three-week-old birds and some in birds up to 17 weeks of age. In most of the standard turkey flocks the mortality was less than 10 per cent, but it was above 30 per cent in nearly 20 per cent of the outbreaks. In the breeding flocks, the average mortality was 60.2 per cent. The size of the flocks, the sex of the birds and the age at which the first clinical signs appeared did not seem to influence the mortality.

Genetic variation in susceptibility to Trypanoplasma borreli infection in common carp (Cyprinus carpio L.).

Gynogenetic reproduction of homozygous females, or crossbreeding two homozygous animals, results in fish lines without genetic variation. Hybrid crosses are expected to express a more stable development than homozygous lines, the latter may have an important value for gaining insight into genetic components of host resistance to parasite infection. We examined the antibody response of carp (Cyprinus carpio L.) to infection with Trypanoplasma borreli. Outbred carp responded with a production of specific antibodies, but highly susceptible isogenic hybrid carp did not. This suggests an apparent relationship between susceptibility and the lack of specific antibody production. This relation was partially confirmed by the passive transfer of immunity with immune plasma. In addition, two isogenic homozygous carp lines were highly susceptible to the trypanoplasm (100% mortality), in contrast with outbred carp, the majority of which survived infection. None of the carp in either homozygous carp line produced an antibody response to parasite-unrelated antigen (DNP-KLH). This suggests that the low antibody response was not entirely due to a poor state of health, but that these carp have a genetically predetermined low antibody response.

Increased susceptibility of bobwhites (Colinus virginianus) to Histomonas meleagridis after exposure.

Bobwhites given heterakid eggs but no Sevin became infected with cecal histomonads, but there was no pathological histomoniasis. Quail given 50 microgram of Sevin (10 microgram/day) behaved normally, but at necropsy they had slightly discolored livers. Quail given various doses of heterakid eggs and Sevin (Sevin increasing from 2.5 to 50 microgram) and those given various doses of heterakid eggs and 10 microgram/day of Sevin developed pathological histomoniasis and mortality rates of 36 and 63%, respectively.

Hexamita meleagridis (Spironucleus meleagridis) infection in chukar partridges associated with high mortality and intracellular trophozoites.

An outbreak of infectious catarrhal enteritis, associated with the flagellated protozoan Spironucleus meleagridis (syn. Hexamita meleagridis), is reported from a commercial flock of chukar partridges in California. The disease affected birds between the ages of 4 and 6 wk and resulted in diarrhea, listlessness, depression, and high mortality. Concurrent infection with other intestinal pathogens, including Cryptosporidia, group E Salmonella, long-segmented filamentous microorganisms (LSFMOs), and Rotavirus-like virus particles, was found in some but not all affected birds. Dermatitis of the face, shanks, and feet, suggestive of B-complex vitamin deficiency, was present in most affected birds as well. Flagellated protozoan parasites could be found in the lumen of the duodenum and jejunum and in the intestinal crypts. In some cases the flagellates were wedged between epithelial cells or were located intracellularly within cells of the mucosal epithelium and the intestinal lamina propria.

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus.

Atlantic salmon Salmo salar L. (Salmonidae) were experimentally infected with Spironucleus barkhanus (Diplomonadida: Hexamitidae). Parasites were found in the blood 1 to 8 wk after infection, after which they disappeared from the blood and were found mainly in the internal organs (e.g. spleen and liver), eye socket or muscles. Mortality (38 out of 40 infected fish) occurred when fish had lesions in internal organs and/or on the body surface. Uninfected fish cohabiting with infected fish became infected after 4 wk, indicating direct transmission. There was no difference in susceptibility to spironucleosis between 3 different families of Atlantic salmon. All families developed the disease with a similar pattern of parasitaemia in the blood, similar clinical signs and gross pathology, and with very high mortality (29 out of 30). Clinical signs of systemic spironucleosis may include anemia, skin blisters, muscle ulcerations or unilateral exophthalmia. Gross pathologies include hemorrhaging of internal organs, splenomegaly or deformed (globulated) spleen, or granulomatous lesions in the spleen and liver.

Fatal Neospora caninum infection in kittens.

Three 3-day-old kittens were inoculated subcutaneously and orally with Neospora caninum tachyzoites. A littermate and the queen were not inoculated with N. caninum and served as controls. Kitten 1 died between 14 and 16 days postinoculation (DPI) and was eaten by the mother. Kitten 2 died 17 DPI and kitten 3 was euthanized 29 DPI in a moribund condition. The control littermate and the dam remained healthy. Granulomatous skeletal myositis and nonsuppurative encephalomyelitis were the main lesions and were associated with numerous N. caninum tachyzoites in kittens 2 and 3. Cysts were found in kitten 3. Oocysts were not found in any cats. Neither lesions nor parasites were found in control cats.

Neospora caninum-like protozoon associated with fatal myelitis in newborn calves.

Neospora caninum-like organisms were found in histologic sections of spinal cord of 4 paralyzed calves as reported by Parish et al. (1987). Tachyzoites divided by endodyogeny. Tissue cysts were up to 62 microns wide and the cyst wall was up to 2.5 microns thick. The organism in calves was structurally distinct from Toxoplasma gondii and Sarcocystis species and reacted positively with anti-N. caninum serum in an immunoperoxidase test.

Mortality of young-of-the-year rainbow smelt (Osmerus mordax) in Lake Erie associated with the occurrence of Glugea hertwigi.

An extensive mortality of young-of-the-year rainbow smelt (Osmerus mordax) occurred in Lake Erie during the fall of 1969. Dead and dying smelt were observed along the north shore from west of Long Point in the central basin to Port Maitland in the eastern basin. No other fish species was involved. Glugea hertwigi, a microsporidan parasite, was observed in 90% of the distressed smelt examined, and is believed to have been a major contributing factor to the mortality.

Mortality in Atlantic salmon (Salmo salar) associated with trichodinid ciliates.

A protozoan infection (Trichodina truttae) was identified in captive Atlantic salmon (Salmo salar) kelts that died in spring of 1988 and 1989. Fish with intense infections showed signs of listlessness, erratic swimming and inappetence. The infection induced excessive mucus secretion, epithelial sloughing and lesions that probably permitted entry of opportunistic bacteria which eventually caused ulcers and death. A seawater bath for 30 min each week for 4 wk effectively controlled the parasite.

Mortality in muscovy ducks (Cairina moschata) caused by Haemoproteus infection.

Haemoproteus infection was confirmed in white Pekin ducks (Anas platyrhynchos domesticus) following placement in a pond where severe mortality from a respiratory condition apparently caused by schizonts of Haemoproteus had occurred in muscovy ducks (Cairina moschata). Blood from one infected white Pekin duck was experimentally inoculated into five muscovy ducks; two of the five developed large numbers of schizonts in the endothelial cells and died. Illness did not occur in the white Pekin ducks.

Newly recognized fatal protozoan disease of dogs.

Histologic sections and case histories from 23 dogs with proven fatal toxoplasmosis-like illness at the Angell Memorial Animal Hospital were reviewed. Toxoplasma gondii was identified in 13 dogs. A newly identified parasite, Neospora caninum, structurally distinct from T gondii, was found in 10 dogs. The newly discovered organism, belonging to a new genus and new species, formed meronts in many tissues of the dogs, especially the brain and spinal cord. Neospora caninum was located directly in the host cell cytoplasm without a parasitophorous vacuole; it divided by endodyogeny, contained more than 11 rhoptries, and did not react with the anti-T gondii serum in the immunoperoxidase test. Meningoencephalomyelitis and myositis were the main lesions associated with N caninum. Ulcerative and fistulous dermatitis was the major lesion in 1 dog.

Cryptobia iubilans infection in juvenile discus.

Four commercial producers of discus (Symphysodon aequifasciatus) were found to have fish infested with the flagellate Cryptobia iubilans. Affected fish had granulomatous gastritis, and many also had granulomatous disease of other organs. The parasite had to be differentiated from the related flagellates Spironucleus spp, which induce different lesions. Transmission electron microscopy was found to be useful in detecting and identifying the parasite. Morbidity and mortality rates in the various fish populations appeared to be linked to a number of variables, including water quality, presence of other parasites and bacteria, diet, species, size, and age of the fish, and optimization of husbandry appeared to be important in alleviating the severity of disease. Metronidazole was not effective for treatment of C iubilans, but bath treatments with dimetridazole (80 mg/L for 24 hours, repeated daily for 3 days) or 2-amino-5-nitrothiazol (10 mg/L for 24 hours, repeated daily for 3 days) may be useful in decreasing the prevalence of infestation.

Hexamitiasis in cadmium-exposed mice.

Mortality was observed in 4- to 5-week-old Swiss Webster mice exposed to 300 or 3 ppm cadmium as cadmium chloride in the drinking water. Mice receiving 300 ppm cadium suffered 26% mortality as compared with 7% of those on the low cadmium dose. Death did not occur in control mice. Clinical signs and histopathology established Hexamita muris as the causative agent. Cadmium lesions were not observed. It is suggested that mortality due to hexamitiasis resulted from synergism between cadmium and H muris.

Deaths in Australian freshwater fishes associated with Chilodonella hexasticha infection.

The cause of the deaths of bony bream and other native fish in the Finke River near Alice Springs in winter 1984 was infection with the protozoan ciliate Chilodonella hexasticha. The parasites induced severe generalised epithelial hyperplasia in the gills, which would have compromised respiratory exchange and killed the fish through hypoxaemia. It is suggested that similar winter epizootics of ectoparasites have contributed to previous deaths of fish observed in the Finke River, in association with dry season conditions of low water temperatures and flow rates.