Prevention of shedding and re-shedding of Toxoplasma gondii oocysts in experimentally infected cats treated with oral Clindamycin: a preliminary study

This work aimed to evaluate the effects of preventive oral Clindamycin in cats infected with Toxoplasma gondii. Twelve short hair cats were divided into two groups (group 1 and group 2). No titres of T. gondii antibodies were detected in these cats before the experiment. The animals from group 1 were infected with tissue cysts of T. gondii and group 2 were infected and treated with Clindamycin (20 mg/kg/day). The infection was done with almost 40-50 tissue cysts for each cat on day 0. The cats from group 2 were treated with Clindamycin by oral rout for 24 days (from day -3 to day 21). At day 45, the groups 1 and 2 were divided into two subgroups with three animals each. Subgroups 1A and 2A were immunosuppressed with dexamethasone (1 mg/kg/day) for30 days and subgroups 1B and 2B were not immunosuppressed. Faecal exam looking for oocyst shedding was made by 30 days after T. gondii infection, and for 30 days after immunosuppression. All kittens from group 1 shedding oocysts after infection, while animals from group 2 did not shed. After immunosuppression period, all animals from group 1A re-shed oocysts and animals from group 2A remained without shed. However, 2 (66.6%) of the kittens from subgroup 2B shed oocysts 19-20 days after re-challenge. Based on this preliminary study, Clindamycin had a complete inhibitory effect on shedding of oocysts by cats, even under severe immunosuppression, which is a new finding not reported elsewhere.     

雏鸡感染堆型艾美耳球虫及体内一氧化氮的生成

本研究利用Griess反应监测了雏鸡感染堆型艾美耳球虫(Eimeriaacervulina)后血浆NO     

Evidence of interaction between Ascaris suum and Metastrongylus apri in experimentally infected pigs

A study has been carried out with the aim to determine possible interactions between Ascaris suum and Metastrongylus apri under experimentally infected pigs. Twenty-eight Iberian pigs were allocated into four groups. Group 1 was inoculated with 5000 infective A. suum eggs; group 2 received concurrently 5000 infective A. suum eggs and 5000 infective M. apri larvae; group 3 received 5000 infective M. apri larvae; group 4 served as uninfected controls. In each group, pigs were necropsied on day 7 (n = 4) and day 28 (n = 3) post-infection (p.i.). Pigs with single M. apri infections showed earlier and more severe respiratory symptoms compared to pigs with mixed infection, while no clinical signs were observed in pigs single infected with A. suum. Mean burdens of immature A. suum and immature and adult M. apri were reduced in pigs with concomitant infection both on day 7 and 28 p.i., respectively. In contrast, the number of white spots was significantly increased on day 7 in pigs with mixed infection. In addition, pigs of group 1 showed the highest eosinophil levels in blood compared to pigs in groups 2 (intermediate levels) and 3 (moderate levels). The results suggest an antagonistic interaction between A. suum and M. apri in concomitantly infected pigs.     

雏鸡柔嫩艾美耳球虫感染与体内NO的生成

本实验研究了雏鸡柔嫩艾美耳球虫(Eimeria tenella)感染与其体内一氧化氮生成的剂量-效应关系和时间-效应关系。在预试基础上，1日龄黄羽肉用雏鸡饲喂基础日粮2周后，随机分成对照组和0.5×10     

Toxoplasma gondii Infections in Chickens (Gallus domesticus): Prevalence,Clinical Disease,Diagnosis and Public Health Significance

Chickens are considered one of the most important hosts in the epidemiology of Toxoplasma gondii infection because they are an efficient source of infection for cats that excrete the environmentally resistant oocysts and because humans may become infected with this parasite after eating undercooked infected chicken meat. The objective of this study is to review worldwide prevalence of T. gondii infection in chickens and to assess the role of infected chickens in the epidemiology of toxoplasmosis in humans. A very high prevalence of the parasite was found in chickens raised in backyards (up to 100%) and free‐range organic (30–50%) establishments.     

Detection of Oocyst‐Associated Toxoplasmosis in Swine from Southern Chile

Pork has been traditionally considered an important source of human Toxoplasma gondii infection. Pigs, as other meat‐producing animals, can become infected by the ingestion of oocysts that are shed in the environment by infected cats or by the consumption of cysts present in tissues of infected mammals, commonly small rodents. The objective of this study was to investigate the level of T. gondii infection in swine from southern Chile that can be associated with the ingestion of oocysts and therefore exposure to a contaminated environment. A total of 340 serum samples from swine were obtained from three commercial slaughterhouses located in the Araucania and Los Rios Regions from southern Chile. Study animals originated from local farms, mainly small commercial producers, and the meat is sold locally. Overall, 8.8% (30/340) of the samples showed T. gondii‐specific IgG antibodies. Of these sero‐positive animals, 80% (24/30) were also positive for antibodies specific against the oocyst stage of the parasite, indicating that animals had been infected recently by the ingestion of oocysts. The observed results suggest a high level of environmental contamination with oocysts on the farms of origin. In addition to the food safety problems associated with the consumption of meat from infected animals, the high level of environmental contamination on the farm represents a direct health risk for people living and/or working on these farms. Consequently, there is a need to develop on‐farm monitoring programmes and identify risk reduction strategies (food storage, water purification, rodent control and contact with cats) that are appropriate and cost‐effective for informal and outdoor type of farms.     

Radiofrequency‐Induced Thermal Inactivation of Toxoplasma gondii Oocysts in Water

Toxoplasma gondii, a ubiquitous parasitic protozoan, is emerging as an aquatic biological pollutant. Infections can result from drinking water contaminated with environmentally resistant oocysts. However, recommendations regarding water treatment for oocyst inactivation have not been established. In this study, the physical method of radiofrequency (RF) power was evaluated for its ability to inactivate T. gondii oocysts in water. Oocysts were exposed to various RF energy levels to induce 50, 55, 60, 70 and 80°C temperatures maintained for 1 min. Post‐treatment oocyst viability was determined by mouse bioassay with serology, immunohistochemistry and in vitro parasite isolation to confirm T. gondii infections in mice. None of the mice inoculated with oocysts treated with RF‐induced temperatures of ≥60°C in an initial experiment became infected; however, there was incomplete oocyst activation in subsequent experiments conducted under similar conditions. These results indicate that T. gondii oocysts may not always be inactivated when exposed to a minimum of 60°C for 1 min. The impact of factors such as water heating time, cooling time and the volume of water treated must be considered when evaluating the efficacy of RF power for oocyst inactivation.     

Amprolium and furazolidone as preventive treatment for intestinal coccidiosis of piglets

Two coccidiostats, amprolium and furazolidone, were used as preventive treatments for intestinal coccidiosis in three-day-old piglets experimentally infected with 50,000 sporulated oocysts of Isospora suis.

All infected piglets, treated or not, displayed clinical signs compatible with coccidiosis. Diarrhea and anorexia appeared around five days postinoculation in the non-treated and in the amprolium-treated groups; these signs were delayed to days 7 and 8 postinoculation in the furazolidone-treated group. The treatments did not prevent growth retardation. Amprolium seemed to reduce oocyst shedding whereas furazolidone had no effect. Villous atrophy was present in all infected piglets.

     

Infectivity of a novel type of Cryptosporidium andersoni to laboratory mice

Previously, we reported 'a novel type' of Cryptosporidium andersoni detected from cattle in Japan, and showed that the isolate was infective to mice. In the present study, we examined the patterns of oocyst shedding in both immunocompromised and immunocompetent mice, as well as pathological lesions in the infected mice. After oral inoculation with 1 x 10(6) oocysts, all five severe combined immunodeficiency (SCID) mice began to shed endogenously produced oocysts on day 6 post-inoculation (p.i.). The number of oocysts per day (OPD) reached 1 x 10(6) on day 17 p.i., and an OPD level of 1 x 10(6) to 10(7) was maintained until 91 days p.i. when the mice were sacrificed. In the five immunocompetent mice inoculated with 1 x 10(6) oocysts, the pre-patent and patent periods were 6 and 19 days, respectively, and the maximal OPD level was 1.5 x 10(5) on average. On histological examinations of infected SCID mice, a large number of parasites were present on the surface of the gastric glands of the stomach, but not in other organs examined. In conclusion, the novel type of C. andersoni, which genetically coincides with C. andersoni reported in other countries, is infective to mice, but susceptibility was lower than that of Cryptosporidium muris infecting rodents from the perspective of infectivity to immunocompetent mice.     

Unusual biphasic disease in domestic pigeons (Columba livia f. domestica) following experimental infection with Sarcocystis calchasi

A novel Sarcocystis species has recently been reported in the domestic pigeon (Columba livia f. domestica) as intermediate host, causing severe central nervous signs similar to Paramyxovirus-1 or Salmonella Typhimurium var. cop. infection. Transmission of the parasite via the northern goshawk (Accipiter gentilis) as definitive host has been established. Experimental infection of domestic pigeons with sporocysts excreted by experimentally infected northern goshawks reproduced the natural infection in the pigeon, proving the causative role of the parasite in the disease. Here, we describe in greater detail the course of the fulminant biphasic disease depending on the infectious dose. Pigeons infected with 10(3) or 10(4) sporocysts showed clinical signs of polyuria and apathy around 10-11 days postinfection (dpi) and sudden neurological signs 51-57 dpi as a second phase of disease. Pigeons infected with higher doses died within 7-12 dpi, also showing polyuria and apathy but without nervous signs. At necropsy, livers and spleens had multifocal necroses and infestations with parasitic stages, namely, schizonts. Moreover, lesions and schizonts were also found in the lung, bone marrow, and next to blood vessels in the connective tissue of various organs. Pigeons infected with 102 sporocysts remained symptomless until 58-65 dpi, when sudden central nervous signs occurred. Major histopathologic findings of pigeons with neurological signs were encephalitis and myositis of virtually every skeletal muscle with high infestations of sarcocysts. Only mild myocarditis and very few cysts were found in the heart muscles. Importantly, a sentinel pigeon developed identical lesions when compared to those of low-dose infected pigeons, suggesting a risk of mechanical transmission of sporocysts from freshly infected to uninfected pigeons in a flock. By contrast, chickens failed to develop any clinical signs or pathologic lesions in the same experiment. The findings further characterize the new highly pathogenic disease in domestic pigeons, which clinically mimics paramyxovirosis and salmonellosis in both phases of the disease and exclude chickens as further intermediate host species.     

Concurrent Ascaris suum and Oesophagostomum dentatum infections in pigs.

The aim of this study was to examine interactions between Ascaris suum and Oesophagostomum dentatum infections in pigs with regard to population dynamics of the worms such as recovery, location and length; and host reactions such as weight gain, pathological changes in the liver and immune response. Seventy-two helminth-na?ve pigs were allocated into four groups. Group A was inoculated twice weekly with 10000 O. dentatum larvae for 8 weeks and subsequently challenge-infected with 1000 A. suum eggs, while Group B was infected with only 1000 A. suum eggs; Group C was inoculated twice weekly with 500 A. suum eggs for 8 weeks and subsequently challenge-infected with 5000 O. dentatum larvae, whereas Group D was given only 5000 O. dentatum larvae. All trickle infections continued until slaughter. Twelve pigs from Group A and B were slaughtered 10 days post challenge infection (p.c.i.) and the remaining 12 pigs from the each of the four groups were slaughtered 28 days p.c.i.. No clinical signs of parasitism were observed. The total worm burdens and the distributions of the challenge infection species were not influenced by previous primary trickle-infections with the heterologous species. Until day 10 p.c.i. the ELISA response between A. suum antigen and sera from the O. dentatum trickle infected pigs (Group A) pigs were significantly higher compared to the uninfected Group B. This was correlated with a significantly higher number of white spots on the liver surface both on Day 10 and 28 p.c.i. in Group A compared to Group B. The mean length of the adult O. dentatum worms was significantly reduced in the A. suum trickle infected group compared to the control group. These results indicate low level of interaction between the two parasite species investigated.     

Pathogenicity of two strains of Newcastle disease virus in the grey‐breasted helmet guinea fowl

Summary

Thirty‐five 6‐week‐old guinea fowl keets, seronegative for maternal antibodies to Newcastle disease virus, were infected with Hens strain (33/56) and Kumarov strain of Newcastle disease virus intramucularly (IM) or intranasally (IN).

Clinical signs were first noticed four days post infection (PI) in the group infected al but five days PI in the group infected IN with Hens strain of Newcastle disease virus. These clinical signs were similar in both groups and included anorexia, droopiness, huddling together, greenish diarrhoea and marked cachexia. Prominent nervous signs, including spasms of the head and neck, were observed in groups infected with Hens strain.

The major gross lesions observed were emaciation with prominent keel bone, empty intestinal tract and distended gall bladder in most keets.

The histological lesions were characterised by meningoencephalitis, necrosis and loss of lymphocytes from splenic and lymphoid aggregates. There was muscular degeneration and necrosis in the gizzard and mild pulmonary congestion and oedema in some keets.

Neither gross or microscopic lesions were observed in keels that had received the Kumarov strain.     

Effect of resinous extract from Commiphora swynnertonii (Burrt) on experimental coccidial infection in chickens

A crude resinous extract from Commiphora swynnertonii was tested against an experimental coccidial infection in local chickens. A total of 80 growing chickens were randomly assigned into five groups, which received different treatments. Chickens in G1 were not infected with coccidian oocysts and therefore served as a negative control. All chickens in G2, G3, G4 and G5 were infected through oral administration of coccidian oocysts suspension at a dosed rate of 1.5?×?10⁴ Eimeria spp. oocysts per bird. Starting from day 3 post-infection (p.i), chickens in different groups were treated for 7 consecutive days as follows: G1 and G2 (positive control) received 5 ml of normal saline as placebo, G3 and G4 were given the extract at 400 and 800 mg/kg bodyweight whereas G5 received anticoccidial drug. Clinical signs, bodyweights, oocysts counts and mortality rates were observed regularly. Results showed that oral administration of the resinous extract to chickens with coccidiosis significantly reduced mortality rate from 94 to 25 % and oocysts counts from 1.03?×?10⁵ to 6.55?×?10³ oocysts/g faeces (p?<?0.05). Also a body condition score chart indicated less severe clinical signs of the disease in the groups which received the extract. Mean daily body weights were slightly reduced by the administration of the extract but this effect disappeared by day 7 p.i. These findings clearly indicate that resinous extract from C. swynnertonii has significant anticoccidial effect against experimental Eimeria spp. infection in chickens. A larger field trial to validate the use of the extract in chickens naturally infected with Eimeria spp. is required.     

The pathogenesis of experimental infections of Cryptosporidium muris (strain RN 66) in outbred nude mice.

Three groups of six-week-old nude outbred mice were orally infected with 400, 20,000 and 1,000,000 oocysts of Cryptosporidium muris (strain RN 66) per mouse, respectively. Oocysts were detected in the faeces from 10-18 days post-infection (p.i.) and continued to be shed in large numbers in all groups until the termination of the trial on day 89 p.i. Clinical signs were not observed in any of the infected mice and there was no significant effect on weight gain compared to uninfected controls. Histological examination revealed the presence of parasites confined to the glandular stomach. Parasitised gastric glands were dilated, hypertrophied and filled with numerous parasites. The glands had lost their normal cellular architecture and were lined with many undifferentiated cells. In some mice receiving the largest innoculum, the glandular mucosa was congested and the lamina propria infiltrated with eosinophils, polymorphs and lymphocytes.     

Immunohistochemical and polymerase chain reaction studies in Neospora caninum experimentally infected broiler chicken embryonated eggs

The diagnostic characteristics of immunohistochemistry (IHC) and polymerase chain reaction (PCR) methods were studied in the tissues of broiler chicken embryos experimentally infected by Neospora caninum. An infection with N. caninum NC-1 isolate was conducted in 70 broiler chicken embryonated eggs randomly divided into seven equal groups. After 8 days of incubation, six groups were inoculated with 10, 10(2), 10(3), 10(4), 10(5), and 10(6) doses of tachyzoites/embryonated egg. The 7th group was considered as control. The mortality rate and pathological changes of the dead embryos and hatched chickens up to 60 days old were noticed. Consecutive sections to those used for histopathological examination including the liver, heart, brain, and chorioalantoic (CA) membrane were subjected to IHC. The intensity and distribution of the immunostaining was graded as highly to mildly positive. For PCR procedure, DNA was extracted from 50mg of the tissues and primer pair Np21/Np6 was used for amplification of the Nc-5 gene. The results of the immunosignaling ranged from variable degrees of mild to moderate staining as dark-brown to brown and coarsely to finely granular, mostly within the cytoplasm of infected cells such as the endothelial cells of blood vessels. The parasite aggregation was more predominant in the heart than other tissues. Immunoreactivity for N. caninum antigen was multifocally moderate positive in the heart, liver and CA of the 10(3) dose, and also heart, liver, brain and CA of the 10(4) dose. IHC showed mildly positive in the liver and heart of the chicken embryos infected with 10 and 10(2) tachyzoites, as well. The results of the PCR confirmed the existence of the parasite in all of the examined tissues from the 10(3) and 10(4) doses. In conclusion, the results indicate a good agreement between IHC and PCR in diagnosis of neospora antigen in the infected tissues.     

应用巢式PCR-杂交技术检出人血样及动物排泄物中弓形虫DNA

弓形虫是一种专性细胞内寄生的原虫，引起人兽共患疾病-弓形虫病。本文报道了一种基于PCR技术检测宠物弓形虫的方法，该方法的特点是通过检测宠物排泄物中是否存在弓形虫包囊，从而诊断宠物是否为真实的传染源。由于宠物的排泄物容易采集，避免了复杂的采样程序，方便于检测。     

Infectivity of Cryptosporidium sp isolated from wild mice for calves and mice

A study was conducted to determine the incidence of cryptosporidiosis in wild mice (Mus musculus) and the infectivity of oocysts from their feces for susceptible calves. The presence of oocysts and the duration of shedding of oocysts in the feces were evaluated in 115 wild mice. Approximately 30% of the mice shed Cryptosporidium sp oocysts, without evidence of clinical infection; recurrence of oocyst shedding was found in about 50% of the mice. Oocysts from the feces of naturally infected mice were infective for calves and mice. Calves began shedding oocysts at 7 days and shed oocysts for about 10 days. Nonfatal, clinical cryptosporidiosis developed in 7 infected calves. The mice began shedding oocysts at 6 days and shed oocysts for 12 days. Fatalities or clinical infection did not develop in 5 infected mice. The results indicated that Cryptosporidium-infected wild mice may be a source of cryptosporidiosis in susceptible calves.     

The pathological changes caused by Eimeria falciformis var pragensis in mice.

Groups of Swiss white mice weighing 25-28 grams were infected orally with 500, 2,000, 5,000 or 20,000 oocysts of Eimeria falciformis var pragensis. Depression, anorexia, weight loss, diarrhea or dysentery, and dehydration were most pronounced at eight to ten days postinfection. The highest mortality, 31%, occurred in mice infected with 20,000 oocysts. None of the mice infected with 500 oocysts died. The pathological findings were equally severe in mice infected with 5,000 and 20,000 oocysts. The enteric lesions, most pronounced at eight to ten days postinfection, were restricted mainly to the large intestine and consisted initially of both cryptal and absorptive epithelial cell destruction and submucosal edema. These changes were followed in 12 to 24 hours by a transient influx of neutrophils into the lamina propria followed by mononuclear cell infiltration which lasted for five to ten days. As the infective dose decreased, the inflammatory response occurred later and was less extensive. When seen, hemorrhage occurred seven to 11 days postinfection. In 50% of the mice infected with 5,000 and 20,000 oocysts, varying degrees of a nonselective mucosal necrosis were seen at eight to 12 days postinfection. In mice infected with 500 oocysts, mucosal destruction was restricted to the epithelium. Neutrophils predominated when necrosis was extensive, otherwise, mononuclear cells were the main inflammatory cells. Two to three days following necrosis, crypt hyperplasia was marked and mucosal integrity was restored. Ulcers, some of which extended into the submucosa, healed by days 14 to 20. Localized granulomatous colitis, induced by trapped oocysts within the lamina propria, was seen until the experiment was terminated at 25 days postinfection. Infection was followed by lymphoid hyperplasia in the lymph nodes and the spleen.     

Experimental infection of dogs (Canis familiaris) with sporulated oocysts of Neospora caninum

Neospora caninum is widely distributed in the world and this parasite is one of the major causes of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum and several species of domestic and wild animals are intermediate hosts. Dogs can become infected by the ingestion of tissues containing cysts and then excrete oocysts. It is not yet known whether sporulated oocysts are able to induce a patent infection in dogs, i.e. a shedding of N. caninum oocysts in feces. The objective of this study was to experimentally examine the infection of dogs by sporulated oocysts. The oocysts used in the experiment were obtained by feeding dogs with brain of buffaloes (Bubalus bubalis) positive for anti-N. caninum antibodies by indirect fluorescent antibody test (IFAT ≥200). Oocysts shed by these dogs were confirmed to be N. caninum by molecular methods and by bioassay in gerbils, and sporulated N. caninum oocysts were used for the oral infection of four dogs. The dogs were 8 weeks old and negative for antibodies to N. caninum and Toxoplasma gondii. Dogs 1 and 4 received an inoculum of 10,000 sporulated oocysts each; dog 2 an inoculum of 5000 sporulated oocysts and dog 3 received 1000 sporulated oocysts of N. caninum. The total feces excreted by these dogs were collected and examined daily for a period of 30 days. No oocysts were found in their feces. The dogs were monitored monthly for a 6-month period to observe a possible seroconversion and when this occurred the animals were eliminated from the experiment. Dogs 1 and 4 seroconverted 1 month after the infection with titer, in the IFAT, of 1600 and 800, respectively; the other two dogs presented no seroconvertion during the 6-month period. Dogs 1 and 2 were euthanized 180 days after infection and were examined for the detection of N. caninum in tissues (brain, muscle, lymph node, liver, lung, heart and bone marrow) by immunohistochemistry and PCR with negative results in both techniques. Bioassay in gerbils with brain of these dogs was also performed and again the results were negative. In conclusion, dogs infected with sporulated oocysts of N. caninum were not able to shed oocysts in feces. However, a higher dose of infection stimulated the production of antibodies against N. caninum in the dogs.     

Comparative efficacy of flubendazole chewable tablets and a tablet combination of febantel, pyrantel embonate and praziquantel against Trichuris vulpis in experimentally infected dogs

Fourteen of 23 dogs developing patent Trichuris vulpis infections by 120 days p.i. with 5000 embryonated eggs were allocated into three groups. One group was treated with flubendazole 220 mg chewable tablets (Flubenol) at the recommended dose regimen once daily for 3 days. The second group was given the recommended single treatment with a tablet containing 150 mg febantel, 144 mg pyrantel embonate and 50 mg praziquantel in combination (Drontal Plus). The third group remained untreated. All dogs were necropsied for worm counts 10 or 11 days after (first) treatment. No worms were recovered from the flubendazole treated dogs resulting in a significant worm count reduction of 100%. In contrast, 2 of 5 animals treated with the combination of febantel, pyrantel embonate and praziquantel remained infected; the geometric mean worm burden was reduced by 99.4% as compared to the control group but did not differ significantly from those of the controls.